Journal of Japanese Society for Extremophiles (2022) Vol.20

ISSN 2186-9936

Sep. 2022

極限環境
生物学会誌

第 21 巻第 1 号

Journal of Japanese Society for Extremophiles

目次
巻頭言‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 2

研究最前線‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 4

MINI REVEW‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 6

ORIGINAL PAPER ‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 15

論文執筆要領‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 72

学会会則 ・ 細則‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 73

運営体制‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥‥ 77

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2

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)	  ところで、筆者は極限環境生物学会のような小規模で専門性の高い学会のほかに、日本化学会や日本農芸化学会のような、より広範な専門分野をカバーする大規模な学会にも所属している。学会によって出会える研究者が異なるため、得られるものが違ってくるからである。決して大規模な学会がメインで、小規模な学会がサブであると考えているわけではない。小規模な学会をメイン、大規模な学会をサブとする研究者もいらっしゃるであろうし、筆者の場合、研究テーマによりメインとなる学会が異なってくる。筆者が所属する複数の学会は筆者にとって「縦糸／横糸」の関係にあり、縦糸と横糸がバランスよく織り成されることで美しい布地が生み出されるのである。従って、決して極限環境生物学会だけですべてを完結させる必要はないと考えている。それぞれの研究者によって、極限環境生物学会の位置付けは異なっていても構わないが、縦糸か横糸のどちらかに選んでいただけるような学会をめざし、副会長としての責務を果たしていきたい。
Reference
Wakai, H., Nakamura, S.*, Kawasaki, H., Takada,K., Mizutani, S., Aono, R., and Horikoshi, K. 1997.
Cloning and sequencing of the gene encoding thecell surface glycoprotein of Haloarcula japonica
strain TR-1. Extremophiles, １(1): 29-35.

3

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)研究最前線	 	 私 の 極 限 環 境 生 物 研 究 	 	
 研究室の集合写真（通称「プロセス研究室」の P のポーズをしている。筆者は後列右端）

富山県立大学工学部生物工学科	  金井保
は じめ に

市仁江海岸）からもほど近い位置にある。この様に本県2021 年 4 月に学生時代より 30 年近く在籍した京都大は極限環境生物研究において魅力的な立地でありなが学大学院工学研究科を離れ、本学に赴任した。現在は、ら、本県を含む北陸地方在住の研究者は少ない印象が富山県立大学工学部生物工学科の応用生物プロセスある。現在は富山県内からの（超）好熱菌の分離なども学講座（写真）に属し、牧野祥嗣講師と戸田弘講師と共開始しているが、本稿では私がこれまでに行ってきた研に研究室を運営している。職場と住まいが同時に変わる究を中心に紹介したい。

のは大学入学以来であり、様々な環境変化に戸惑いながらも刺激の多い日々を過ごしている。これまで富山県

１．極 限 環 境 生 物 に お け る 環 境 適 応 メカ ニ ズ ムとのゆかりは特になかったが、実際に住んでみると元がの解明

北陸地方（福井県敦賀市）の出身であるので生活習慣

私は大学 4 年生の卒業研究で「炭化水素資化性酵母

や食生活などで地元との共通点を見出し、懐かしい感

Candida tropicalis の遺伝子プロモーターを用いた外来覚を覚えることもしばしばである。その一方で、3000m 級

遺伝子発現系の構築」というテーマで研究生活をスターの立山連峰や日本海随一の深さを誇る富山湾の存在

ト さ せ た 。 本 菌 は n- ア ル カ ン を 資 化 可 能 ななど、他県にはないダイナミックな自然環境は本県の大nonconventional yeast（極限環境生物？）であり、n-アルきな魅力である。高低差に富む地形をもつ本県には多カン添加時に、細胞内小器官である peroxisome (Ps)が様な自然環境が存在しており、この様な特徴を活かして細胞内に増殖・蓄積する。一方で、本菌をグルコースでいくことも地方公立大学に課せられたミッションの１つと培養すると、Ps や Ps 局在性酵素群の発現誘導が抑えら考えている。さらに本県は、東洋大学の高品先生らによれるという、いわゆる「グルコース（カタボライト）抑制」現

り単離され、三角形の菌としても知られている高度好塩象が起きる。私の研究では、グリオキシル酸回路の鍵酵

性アーキア Haloarcula japonica の単離地（石川県珠洲素である isocitrate lyase (ICL)遺伝子のプロモーター領

4

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)域を用いた誘導型遺伝子発現系の構築を行った。
この研究を通じて、炭素源の違いにより細胞内の代謝系が動的に変化することや、その発現制御の鍵因子として各遺伝子のプロモーター領域が巧妙に機能していることなどを強く意識するようになった。
博士学位の取得後は、同じ専攻に所属していた今中忠行教授（当時）のグループでポスドクとして勤務した。今中研では鹿児島県の小宝島より単離した超好熱性アーキア Thermococcus kodakarensis
KOD1 株を用いて、DNA ポリメラーゼを始めとする有用遺伝子（遺伝子工学関連、糖質分解関連、タ
ンパク質分解関連など）の解析を幅広く行っていた。
その一方で、本菌を始めとする超好熱菌では、遺図 1 Tar の結合配列と制御遺伝子群

伝子組換え系が利用可能な種が、その当時存在せず、超好熱菌の細胞生理学的研究はほとんど行われて い な か っ た 。 そ の 後 、 今 中 研 究 室 に お い て T.

２. 超 好 熱 性 ア ー キ ア の 水 素 発 生 機 構 の 理 解

kodakarensis の遺伝子組換え系が開発されたことから、

とそ の 工 学 的 応 用

本菌の環境適応に関わる転写制御機構の解明に関す

今中研に入って最初に与えられた研究課題は、T.

る研究を開始した。研究の一例として、解糖系と糖新生

kodakarensis のゲノム解析により見つかった細胞質型系の代謝フラックスをコントロールする転写制御因子[NiFe]ヒドロゲナーゼ(Hyh)の大腸菌での異種発現でTgr について紹介する。遺伝子破壊株のトランスクリプ

あった。水素は理想的なエネルギーキャリアとして注目

トーム解析や in vitro でのタンパク質機能解析を進めたを集める反面、その生産は化石燃料に依存し、再生可

結果、Tgr が、環境中の糖濃度を検知するセンサー機

能エネルギーを用いた水素生産法の確立が急務である。

能を有し、さらにはプロモーター領域への結合や解離を生物的水素生産はその解決法になり得る技術であり、通じて解糖系および糖新生系遺伝子群の転写を制御

中でも発酵水素生産は、水素発生速度が高く、スケー

する、多機能性の転写制御因子であることが判明した。

ルアップが容易であるなど、生物的水素生産の中ではTgr により制御される遺伝子群(Tgr regulon)の総数は 30

最も工業化に適した特徴をもつ。

を超え、アーキアで見つかった regulon としては最大級

ヒドロゲナーゼの異種活性発現は、その活性中心構である。この Tgr regulon の発見は、超好熱菌がいかに造の複雑さのため、現在でもほとんど成功例がない。た少ない遺伝子数で環境変化に適応し、生命を維持するだ幸いなことに T. kodakarensis 菌体は、ピルビン酸やデか、という疑問に対する一つの回答とも言える。Tgr 以外

ンプンを基質として培養を行うと高い水素生成能力を示にも、T. kodakarensis の Trp 生合成遺伝子群の転写制すことが判明し、その後、菌体の連続培養による水素連

御に関わる転写調節因子の探索を行った。ゲノム情報

続生産システムを構築した。さらに本菌の水素関連代

より、転写制御因子の候補タンパク質を in vitro で合成

謝に関する遺伝学的研究により、本菌の水素生産に関し、そのライブラリーを用いたスクリーニングにより、Trp

わるヒドロゲナーゼとして、膜型[NiFe]ヒドロゲナーゼ

生合成遺伝子群のプロモーター領域に結合する転写(Mbh)を同定し、一方で Hyh は水素吸収に関与してい制御因子 Tar を同定した。Tar は本菌の芳香族アミノ酸ることが判明した。これらの[NiFe]ヒドロゲナーゼの生理（Trp, Phe, Tyr）の生合成系を共通して制御する、グロー的機能に関する知見を基に、水素高生産株の遺伝子

バルな転写活性化因子である（図 1）。

工学的育種を進め、連続培養時における単位菌体あた5

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)りの水素生産速度として、120 mmol H2 g-dcw−1 h−1 を達を完全に資化可能な KC04ΔtM1 株を創出した。本株の成した。

消費キチンに対する水素生成の割合は、理論値の約85%に達した。

続いて、非可食性バイオマスからの水素生産系の構築を目指し、未利用バイオマスであるキチンに着目した。
T. kodakarensis のゲノム上には、キチン分解・資化遺伝

最後に子群が存在するにも関わらず、キチン依存的な生育は

富山県は北陸随一の工業県であり、生物関連分野で確認されていなかった。これは本菌のキチン分解・資化は製薬企業のプレゼンスが特に大きく、学生の主たる就遺伝子群の発現量が不足しているためであると考え、こ職先となっている。そのため、学生も薬に関わる研究をれらの遺伝子群の強化により、キチン資化が可能な菌好む傾向があるように感じている。この様な環境の中、体を育種できると考えた。そこでキチン分解・資化遺伝

極限環境生物に関する研究の魅力をどのように伝える

子群のプロモーターを高発現プロモーターへと変更しか、日々試行錯誤中である。現在まで私が研究者を続たところ、キチン分解能は向上したが、一方でキチン分けられているのも、恩師を始めとする多くの魅力的な研解産物である glucosamine の蓄積によってメイラード反究者の方々から多くの刺激を受けてきたからであり、私応が生じ、添加したキチンの完全分解には至らなかっ

自身も研究の面白さや醍醐味を一人でも多くの学生にた。そこで adaptive engineering 手法による菌体育種を伝えていきたいと思っている。

組み合わせることで、最終的には添加キチン（0.4% w/v）

	
 6

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)Journal of Japanese Society for Extremophiles (2022) Vol.21(1), 7-12

MINI REVIEW

Katayama T¼ɏñɘ.=&)Œ>+-&# Atribacterota ɂɟőà OP9/JS1ɠǠǻ0ǠǲŮȬ+ƩƧƩĴ
ƪūĺȀǨÞǂǎĸ ïìȞƔƨ÷ǂǎȷɂƟĖ ƈŲ
Institute for Geo-Resources and Environment, Geological Survey of Japan, National Institute ofAdvanced Industrial Science and Technology (AIST), AIST Tsukuba Central 7, 1-1-1 Higashi,
Tsukuba, Ibaraki 305-8567, Japan
Corresponding author: Taiki Katayama, katayama.t@aist.go.jp
Tel: +81-29-849-1537, Fax: +81-29-861-3666Received: Feb. 14, 2022/ Revised: Mar. 14, 2022, Revised: Mar. 24, 2022/ Accepted: Mar. 29, 2022
Abstract

proteins among gram-negative bacteria. Physiologically,

Cultivation of uncultured microorganisms is a

RT761 primarily produces hydrogen for electron

critical step in uncovering their phenotypic features,disposal during sugar fermentation, and co-cultivationsuch as cell structure and metabolic function. However,with a hydrogen-scavenging methanogen improves

due to difficulties in cultivation, there may be more thangrowth. Similar metabolisms that can be associated with

80 candidate phyla in prokaryotes, significantly

methanogens

have

been

outnumbering phyla with cultivated representatives.

cultivation-independent

analyses

Whiledisplaying their syntrophic lifestyle in vairous anoxic

omics-based

cultivation-independent

predictedofinAtribacterota,

ecosystems.

characterization can circumvent cultivation, metabolic
reconstruction is based on genes characterized incultured organisms and, thus, prediction of novel

1 :.

phenotypic features of uncultured microorganisms ĪƩƠɟ*1ÔŦƩƠ0Ǡǻ+ÚǠǻDľɠ

remains challenging. Recently, we have succeeded in1ǰƾ*Ȉ-#:©

0İȟDŒ>.@

isolating a bacterium that affiliates with the candidate#:.1ƨ÷.ąî@¢;0ĪƩƠǋ0

phylum ‘Atribacteria’, and we proposed a new phylum,>ėñï* 12 ǋǊĞ.7*ɍǍ¼ɏ

Atribacterota. The isolate, RT761, is an anaerobicøź

fermentative bacterium that has an intracytoplasmic

ȷ¼1Ʀœƚ*ñɘƆŒ-ǚǣǭ.=&)Ó

membrane apparently surrounding the nucleoid. The:>A)@ǠǻDŘ;ɜŵ0¼ɗɊǞ

RT761 genome encodes proteins with features that(phylum) *¼ɗ@+Ǜ 110 ɂ+-@!0

might be related to the complex cellular structure,%ñɘAİȟȒȤA#ǋá7A@ɂ1Ǜ 3

including N-terminal extensions in proteins involved in

Å.Ȱ-(Genome Taxonomy Database, GTDB,

important processes (such as cell-division protein FtsZ)

https://gtdb.ecogenomic.org/)Ź? 7 Å0ɂɟ«ȃɂɠ

and one of the highest percentages of transmembrane

7

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Journal of Japanese Society for Extremophiles (2022) Vol.21(1)1ñɘDƨ÷>ƺłħ>A#ȴăȸ¾0

ĂŽƨ÷.ũı@Ǡǻ0ǚǣǭ+)ƩĴĆƵ.

8>ŮĵA@śñɘǚǣǭ*@A>1ȽȆȊA->; 10)OP9 +)ȕșA# 1998

ćĉ.ąî@+A@śǀ0ŕɞƠȟɟ]PtĚşǝǖñɘť1ħ>A)-&#ŝǌ*\ɠ.-">ĪƩƠ]Pt\+ä2A)

1ñɘ.=&)Œ>+-&#Ōɂ Atribacterota

@ĪƩƠ]Pt\D¼ɏñɘ)ǘǪ.

(őàAtribacteria’)0ƩƧĆƵƩĴĆƵơī.'ơī@+1ÔŦƩƠ0Ŕȯİ+üůİ!

)ŝǚǣǭ.Ʌ@A7*0ǀȈD-0ŧĝ.

> ŭ Ȗ  #   -   GTDB * 1  ŝ ɂ 1@ȮË0ȰǊDƍƧȎ@$*-ƩƠȞƔ+)Ǉ.řÈƉƫ@*ȽȆ*Caldatribacteriota’+åàA)@ƎDȵ@@#:*1 Atribacterota .ǣ)ȁȒ@ ŘȦǓǯ>1«ȃɂAtribacteria’.ęĔ@ Ǡ ǻ RT761 ť 0 ǝ ǖ ñ ɘ . ĵ Æ   Ō ɂ
AtribacterotaŌĔŌǋ Atribacter laminatus +)åǠ ǲŮȬ+Sgv0ơī

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àńŨ# Atribacterota 11998 Ě.HJW

 2013 Ě.Ƒƅ>¶ŮǕA# OP9 0Sgv

bëǐ³í0ƑƅóǍƠ> 16S rRNA ȴă.1Q{vɉİǻ0Ľ'ûǵ.ɅC@ȴăɟ¥P+)Ū»A«ȃɂ OP9 +åàA#

2LipidA ƩÞĵ.Ʌ@ lpxAɠȈ'&#5)

!0àǉ1ȔŊŀØA#Ƅ Obsidian Pool+>Q{vɉİð0ǵŮȬDř@+ŃĊ.ƭş)@2013 ĚVQ}Y}Sgvw

A# 2)ßů.Ķ\SgvȎš.=&)Ƒƅ> OP9 0Sgvȸ¾

RT761 ť;Q{vŤǹȔɛ.)ɉİDǆ#¶ŮǕAȘųǳ-,ŃĊA# 2)ßœ.

+BRT761 ťǠǲDɑăɖĪɁ*Ȍď#+Ő0ġ#>-ƨ÷.ƭş#+>

Bûǵµǵ(Ǡǲǵ).Ç)Sgv DNAŕ¼ɏñɘ# A. laminatus+ĳã.%-E*OP9 .C?ŖĊɂàDÊ9Ǡǲµǵȕ:>A#(ê 2) 6)P{HKɑă Atribacteria’ + ń Ũ  A # 2)  2016 Ě . 1 ǩbxQ{lGDá9ɖĪɁȌď.=&)

phylogenomic Ȏš.ò()OP9 0Āÿǚǣǭ*RT761 ť10ơīƵ-ǠǲŮȬDǆ#

@ JS1 Atribacteria’.ÇC&#10)

JS1 1Ƌ• Ǡǲµǵ1Ǡǲ¼Ȃ0,0ŻɊ0Ǡǲ.;Sg

ĝóǍƠ0¯Óǚǣǭ+) 2004 Ě.«ȃɂ JS1 +vDÊ9Ģ*ȌďAûǵµǵ+ȭÉ)åàA#ǚǣǭ* @ 14)OP9 + JS1 1¼ăǚǣ¼Ȃ)#Ų*Œƿ.ÌÀA)?¼ɗɊǞ ǧ(class)• 3 ŵ°ǐ¡ŮȬxa}*1Ǡǲµǵ1Ǡǲǵ+~o}*Ʋ-@Atribacteria’.ęĔ@ƨ÷PƢǐ)ąî#7#Ǡǲµǵ0Ą;Ȍď

1ĂŽĿž¸ƧŰw\ƳȺŰƑƅƃƬ

A-&#ƋĝóǍƠ-,ů -ƨ÷.Ū»A(ê 1)Ĝſ-• |r[vůǗă(2030 nm)Ǡǲµǵ0µ¬0ǠǲȟɟSgv DNA +ßǏɄɠ$*-Ǡǲµǵ0û¬0Ǡǲȟ.)ɟđ--
>;ɠȌďA# ÔŦƩƠ0ȮË0Ȍƚ.);Ƿãƍơī+Ȑ@ûǵµǵ.Ç)0ǒ 3 0ǵąîê 2 A. laminatus RT761 ť0ǠǲŮȬ(A) ȩȰðɑăɖĪɁ.=@ȠǾ½Ɵ0·ƽƴ;<;<+Ȉ@Sgv DNA Ǡǲµ0ǵ.Ê
7A#ůăȌďA@(B)P{HKɑăǩb
xQ{lG*ņģ¶ŮǕA#Ǡǲ0W{
HWƮ®ļý#Ʈ®*1ûǵµǵǠǲµǵ+-@ê 1 Atribacterota ɂ 16S rRNA ȴăP0ƨ÷¼ĘSILVA a\oW(138.1 SSU Ref
NR 99, https://www.arb-silva.de/)>OP9 + JS1.!A"AęĔ@Pȸ¾a\DØħƭş@ƨ÷+.ɍȑ# Others .
1 lake sediment, hydrothermal sediment -,á
7A@8

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)するのかどうか、現時点では完全に証明されておらRho）。保存性の高いこれらのタンパク質に NTE

ず、RT761 株の細胞構造について上記以外に 2 通りが認められただけでなく、他の Atribacterota ゲノの解釈が為されている

12)

ムにも NTE が保存されていた。

。1 つは、最外膜は外膜ではなくタンパク質で構成される S 層であり、従って、

	
  ①と②は RT761 株の生体活動において膜機能が中間の膜は外膜、最内膜は内膜(細胞膜)という解釈重要であることを示している。③の NTE は以下のである。これは、クライオ電子顕微鏡観察において、

点から RT761 株の複雑な細胞内構造に関与してい最外膜に局所的に見られる規則的な凹凸構造が、Sると推測された。一部の細菌は細胞内に Bacterial層に似ているからである。クライオ電子顕微鏡を用

microcompartment (BMC)と呼ばれるタンパク質のい、強い電子線を RT761 株の細胞に当てると、脂殻に覆われた多面体構造物を有し、細胞質と物理的質二重層の証である二重線構造が 3 つの膜すべてに隔離させた BMC 内部空間で特定の代謝反応を行に観察される結果は、第 3 の脂質膜が存在すること

うことが知られている（例えば、シアノバクテリアを強く支持する。しかし、そのような「見た目」以

3)。BMC 内部で働く酵素は NTEのカルボキシソーム）外の観点から検証されていないため、S 層である可を有し、その酵素が BMC 内部に局在するために能性を完全に排除できない、というものである。もNTE が不可欠であることが明らかとなっている 7)。

う 1 つは、細胞内膜は内膜とどこかで繋がっており、

RT761 株にて発見された 34 個の NTE タンパク質の従って、細胞内膜は内膜の一部という解釈である。

内、BMC に関連する酵素は BMC ホモログタンパ

トモグラフィーでは、細胞を回転させながら連続的

ク質の遺伝子クラスターに含まれているに撮影し、その情報に基づいて立体構造をモデリン

deoxyribose-phosphate aldolase のみと考えられた。そグするが、360 度回転させることはできない。従っの他のタンパク質の NTE も細胞内局在性を決めるて、立体構造モデルの「死角」において、細胞内膜機能を有するのかどうか、現時点では実証されていが内膜と繋がっている可能性を排除できない。1 点ない。しかし、細胞分裂に必須な FtsZ については目の解釈については、細胞壁であるペプチドグリカ

以下の興味深い特徴が明らかとなった。RT761 株とンの位置を確定させる、あるいは、ペリプラズム空

Atribacterota ゲノムは FtsZ 遺伝子を 2 コピー有して間・内膜・外膜に特異的なタンパク質の局在性を明おり、その内の一方のみが NTE を有する。また、らかにすることが必要と考えられる。2 つ目は、ト

対数増殖期においてどちらの mRNA も高発現してモグラフィーによる細胞観察数を増やす、あるいは、いる。タンパク質二次構造予測において FtsZ の NTE

細胞の完全な立体構造を復元することのできる別は両親媒性のヘリックス構造を形成する。FtsZ と協の顕微鏡技術が必要と考えられる。

調的に機能する FtsA は、C 末端に両親媒性のヘリッ	  どの解釈であれ、RT761 株が従来の細菌とは大きクス構造を有し、これを以て細胞膜に結合すること

く異なる細胞膜構造を有することは間違いない。上で FtsZ リングの足場と為すことが知られている 11)。

記 RT761 株の特異な細胞膜構造を念頭に、全ゲノ

FtsZ と異なり、FtsA 遺伝子は Atribacterota ゲノム

ム配列(約 3.1 Mb)及び発現 RNA を解析したところ、上には 1 コピーしか存在せず、NTE を持たない FtsZ

次の特徴が明らかとなった 6)。

遺伝子と同一オペロン上に並んでいる。以上の in① 対数増殖期において mRNA 発現量トップ 10 のsilico 解析から、細胞分裂において NTE 含有 FtsZ

タンパク質の半分が膜結合型タンパク質であっはもう一方の（一般的な）FtsZ とは異なる細胞局在た(いずれも機能不明)。

性ならびに機能を有すると推測された。既知の細菌② 全タンパク質に対する膜貫通ドメイン含有タン

基準株 6751 株のうち 9 株は RT761 株と同様に NTE

パク質の割合が多く(約 30%)、既知のグラム陰性

含有 FtsZ を有するが、両親媒性のヘリックス構造

菌 3503 株中トップ 10 に含まれた。

が予測された配列はなかった。RT761 株の FtsZ の③ 他のバクテリア系統群の相同タンパク質には無

特徴は、多重の層という特殊な細胞構造を二分裂さ

い、N 末端の余剰配列(N-Terminal Extension、以せるために必要なのかもしれない。第 3 の脂質膜を下 NTE)（約 10〜70 アミノ酸残基）を有するタン

証明し、NTE 含有タンパク質の細胞内局在性を明パク質が 34 個見つかった。その中には、細胞分らかにすることは、多重膜間の輸送システムの解明

裂、DNA 複製、DNA 修復、転写制御、エネルギーにもつながるだろう。

保存といった生命活動の根幹を為すタンパク質が含まれていた（例えば、Cell division protein FtsZ、

生理・生態

DNA polymerase、Transcription termination factor

9Journal of Japanese Society for Extremophiles (2022) Vol.21(1)RT761 ť1Q}TWD¼ȎžǟȹȻ

 RT761 ť1ÎǼƼDĬ.Ĝ@ÑɅŠMWƬȻËƙǟ+ŬĪȿ0J\g}DƩĵ@ǤĐĂ>ñɘA#ÑɅŠMWƬ0þƞMW1ɝŌ

6)ǃȻǃȻ¦ɀ

>ŗŌ)óǍ#ÏƍƋİ0óǍƠĕ0ǄȻDɑăÙč¡+@ĂŽäâ1Ľ#-h_ïĕžɟÚƋžɠ.Ɨą@#%*Ȝư)?

^ñɘ.)Žƻ0žǟƘĞøÇ@+ø6)źɆČA#!*žǟÁƫİw\ƩĵǻƞMWƩƪ.)1ïĕžDï.ƀ8+0´ñɘ.=?Žƻ0žǟDɈÕ@+

MW+žD¼ɏ

RT761 ť0øźȫĞÖ3ǻ¡ĎĞ0ɖǽ-øÇ<ǁɠ0ȷ;ƀ8>AƂǁŰ.Ɩ:>A@

ȕ:>A#ƳȺǻ1ȎǙ.=&)ƩƪA#£Ä

RT761 ť1ȰÕ0ƋĝóǍƠ.ƭş@0Ƃǁ@1řųƠ.ƐRT761 ť0õŰóǍƠ+ïĕžDĂŽƵ.ɍǍñɘ@+.

ÞFY^} CoA >J\g}7*0×ĮDȏ=&)¼ɏA#ïĕž0öƘĞ1Ƌž+ßǊĞā@Ⱥǟ0ȴăDř@'7?J\g}*ƳȺǳDř@.;C>,0=-ñɘ00ĂŽäâ0ƜųɑăÙč¡;ȻËƙǟŞøźlIX*

û1ţƏ)@ 6)w\Ʃĵ.ȳ#ȶ°Ƶ-Žİ0ƳȺǠǻ*0ɑăDžǟ

@&);ƳȺƪƠ+)J\žƗİþƞMWƬ+ä2A)@ėūƵ-þ@0ɌȜưĕ0óǍƠɟƇ@Ƌž+Ȳ&)ǄȻ156Ū»A!

g}D5+E,ƪƩ-0+>

ƨ÷*?ȧ0 RT761 ť0Ș0ơī+ȗ

RT761 ť1.žǟƩĵ.=&)ȻËȶ°h{

æƵ*

@

WDǦĽ@+ǆçA@RT761 ť0ř@ FeFe ñɘDƨ÷>ƺłħ>A# Atribacterotahydrogenase 1ɒǶƳƵ-×Į*ɂǠǻ0Sgvȸ¾Ĳô>1RT761 ť+ß

@ NADH ȻËÖ3mbȶ°DǶƳƵ-×Į*@1Ʋ-@ƲËȘųǳŃĊA)@(1)

@ȶ°ð
@

2013 Ě.VQ}Y}SgvÖ3w\SgvȎš

+.=&)ȶ°ðɑăȱ¡0¶ƩDžǟƩĵ*Dƫ)Ƒƅ>¶ŮǕA# 2 '0Sgv.1

Ferredoxin 0ȻËÖ3mbȶ°+´Ĥ

10)



,%>;VQd}pm^cDř@ Endo-1,4-

A>0ơī>RT761 ť1žǟÁƫİw\Ʃĵ-glucanase Ȉ'&)?Y}W¼ȎǳD

ǻ+0´Ʃ-C%žǟǋɄȱ.ơË#ř@+ŃĊA)@ 2)-RT761 ť1Y}ȘųǳDř@+Ȑ@-RT761 ť1ǄËž

WD¼Ȏ@+*-16S rRNA ȴǟąî*øźɆČA@+>žǟÁƫİ

ă.ò(¼ăǚǣȎš*1RT761 ť+1¼ɗɊ

@(Electron confurcation)

ȝ+Ûǳ*'&)@

2), 10)A1OP9 P~c0ƨ÷Ǟƹ(order)~o}*Ʋ-&)@2 '0Sg

P0üĂŽƌËŰ-,0w\Ʃĵƨv0% 1 '1Y}WDƫ#KUHb*÷>Ū»A)@ƚ(ê 1)+Ŷ.ȗæƵ*0ɍǍñɘDǿ&#ȔŊ>¶ŮǕA)@@

Řȳ-ñɘŞ.@ªÇœɄ 5 Ő(ÒƢñɘ)+7#Y}W¼Ȏ.Ʌ@FugȻŹò+ slow-grower 0 RT761 ť.+&)w\Ʃ§ąA)@+>Y}W¼ȎǳDř

ĵǻ+0ǋɄžǟȱ1ĂŽƨ÷*e_^DƤħ@ÛǳİĠŇĽA@ 2)

@#:0ȽȆ-ķƱ-0*

 2014 Ě . L d ] 0 Ɠ ž  > ¶ Ů Ǖ  A #B

ȁ 1 ƨ÷>¶ŮǕA# Atribacterota ɂSgv0ŃĊȘųǳǚǣǭ

SgvØħA#ƨ÷ŃĊƲËȘųǳ

OP9

Hot springŒ

Meromictic lake´ƩƵȹȻ¼Ȏ

4)

JS1Marine sedimentȎǙJ\g}ƳȺ

1)

JS1

Bioreactor´ƩƵmkKȻ¼Ȏ

10)

JS1

Oil reservoir

ƙËžǟ(mi)¼Ȏ

9)

JS1

Marine sediment(nu)Y}W¼Ȏ
ȎǙJ\g}ƳȺȎǙJ\g}ƳȺ
ȹȻƩĵ

10

ğƫ°
2)

13)

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)Atribacterota 門のゲノムからは、還元的アセチルを駆使して、微生物ダークマターの実態を探ろうとCoA 経路(Wood-Ljungdahl pathway)に関与する全てする研究が近年盛んに行われている。上述の通り

4)の酵素の遺伝子が見つかっている 。同試料の 16S

Atribacterota も 2013 年の研究 2)を皮切りにゲノムかrRNA 遺伝子アンプリコンシーケンスデータのネッ

ら生体機能が推定されてきた。一方で、細菌ゲノム

トワーク解析では、Atribacterota 門の検出率が水素からアノテーションされたタンパク質の約 3〜7 割

利用性メタン生成菌群(Methanomicrobiales 目)と強が機能不明な hypothetical protein であることから、

い正の相関を示したことから、共生的酢酸酸化能、微生物ダークマターの全貌解明には環境ゲノム解

すなわち、酢酸を二酸化炭素に酸化する過程で生成

析だけでは不十分である。実際、培養実験から明らされる水素をメタン生成菌が速やかにメタンに変かになった RT761 株の特徴、すなわち、複雑な細換することでエネルギーを獲得する代謝機能を有胞内構造、水素利用性メタン生成菌との共生は、ゲすると推察されている。メタン発酵槽から再構築さノム配列情報のみでは推定が難しい知見と言える。

れたゲノムからは、メタン生成菌と共生的にプロピ

当分野ではこのような環境ゲノム解析の限界や培オン酸を酸化する代謝機能が推定されている 10)。こ養株から得られる知見とのギャップも認識されてちらは、methylmalonyl-CoA pathway に関与する全て

おり、近年では微生物ダークマターに有効な培養方の酵素の遺伝子が見つかっており、メタトランスク法も開発されつつある 8)。Atribacterota 門について

リプトーム解析にてそれらの遺伝子の発現が確認は、同門の他の系統群(JS1 など)の単離株を更に獲されている。アラスカの油田から再構築された得し、生理・生態・進化・分類学的知見を深めるとAtribacterota 門のゲノムからは、芳香族・脂肪族炭同時に、培養によって明らかとなった表現型を司る化水素の嫌気分解のキーとなる Fumarate addition

遺伝子型を紐付ける等、ゲノム科学の基盤データをreaction を担う酵素遺伝子が検出されている

9)。し拡充することが重要であろう。

かし、β酸化を含む下流の反応を担う遺伝子群は検出されていない。一方、プロパンを発酵的にカプロ

引用文献

ン酸に変換して ATP を獲得する反応経路が提唱さ1)れている。これらの推定代謝反応は油田から得られ

Carr, S. A., Orcutt, B. N., Mandernack, K. W.,
Spear, J. R. (2015) Abundant Atribacteria in

た JS1 クレードのゲノムに特有なことが判明している。

deep marine sediment from the Adélie Basin,	  Atribacterota の各系統群が生息する環境に適応しAntarctica. Front. Microbiol., 6: 872.

た代謝機能を有することは、本門に帰属する 16S

2)rRNA 遺伝子クローンの系統分布が環境分布と概ね

Dodsworth, J. A., Blainey, P. C., Murugapiran,
S. K., Swingley, W. D., Ross, C. A., Tringe, S.

リンクしていること 10)と調和的である。全体で見る

G., Chain, P. S., Scholz, M. B., Lo, C. C.,と、本門はメタン生成菌と共生する可能性を示す点で共通している。RT761 株にて観察された特異な細Raymond, J., Quake, S. R., Hedlund, B. P.

胞構造が OP9・JS1 クレードに共通し、その構造が(2013) Single-cell and metagenomic analyses

共生的な生理生態とどう結びついているのかが今

indicate a fermentative and saccharolytic

後の研究の焦点となるであろう。ただし、上述の再

lifestyle for members of the OP9 lineage. Nat.

構築されたゲノムから得られた情報は、未だ実証さ

Commun., 4: 1854.

れていない点に留意したい。各文献にて記載されて3)いるゲノムビニング及びアノテーションの精度や、

Fan, C., Cheng, S., Liu, Y., Escobar, C. M.,

メタトランスクリプトームやプロテオーム、集積培

Crowley, C. S., Jefferson, R. E., Yeates T. O.,

養実験等、異なる研究手法から得られたサポート

Bobik

データの有無に基づいて尤度を判断し、今後の生態

sequences package proteins into bacterial

研究や分離培養に活用すべきと考えられる。

T.

A.

(2010)

Short

N-terminal

microcompartments. Proc. Nat. Acad. Sci.
USA., 107: 7509-7514.

おわりに
4)

	
  大規模な DNA シーケンス解析技術が広く普及さ

Gies, E. A., Konwar, K.M., Beatty, J. T. ,れてきたことで、メタゲノム、シングルセルゲノム、Hallam, S. J. (2014) Illuminating microbial

メタトランスクリプトームといったオミクス解析

dark matter in meromictic Sakinaw Lake.

11

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)5)

Appl. Environ. Microbiol., 80: 6807-6818.

Cultivation of elusive microbes unearthed

Hugenholtz, P., Pitulle, C., Hershberger, K.L.,

exciting biology. Nat. Commun., 12: 75.
13)Pace, N. R. (1998) Novel division level6)

bacterial diversity in a Yellowstone hot spring.

Pockalny, R., Murray, R. W., Smith, D. C.,

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D’Hondt, S., Orsi, W. D. (2020) Atribacteria

Katayama, T., Nobu, M. K., Kusada, H., Meng,

reproducing over millions of years in the

X-Y., Hosogi, N., Uematsu, K., Yoshioka, H.,

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Kamagata, Y., Tamaki, H. (2020) Isolation of

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Lehman, B. P., Chowdhury, C., Bobik, T. A.

identified by 16S rRNA gene sequences(2017) The N terminus of the PduB protein

originally found in deep marine sediments.

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Appl. Environ. Microbiol., 70: 5708-5713.

theprotein

shell

of

the

Pdu

Lewis, W. H., Tahon, G., Geesink, P., Sousa,

Nat. Rev. Microbiol., 19: 225-240.
Liu, Y-F., Qi, Z-Z., Shou, L-B., Liu, J-F.,
Yang, S-Z., Gu, J-D., Mu, B-Z. (2019)
Anaerobichydrocarbon

degradation

incandidate phylum ‘Atribacteria’ (JS1) inferredfrom genomics. ISME J., 13: 2377-2390.
Nobu, M. K., Dodsworth, J. A., Murugapiran,
S. K., Rinke, C., Gies, E. A., Webster, G.,
Schwientek, P., Kille, P., Parkes, R. J., Sass,
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Journal of Japanese Society for Extremophiles (2022) Vol.21(1)Journal of Japanese Society for Extremophiles (2022) Vol.21(1), 13-23

MINI REVIEW

Inoue K

Photoreceptive membrane protein rhodopsins of Asgard archaea
J^SkJTJ2·äģȈcrVȰkx\ƂĐěƳŏǔǢŘāThe Institute for Solid State Physics, The University of Tokyo, 5-1-5 Kashiwanoha, Kashiwa, Chiba,277-8581, japan
Corresponding author: Keiichi Inoue, inoue@issp.u-tokyo.ac.jp
Phone:+81-4-7136-3230
Received: Feb. 14, 2022/ Revised: Mar. 30, 2022/ Accepted: Apr. 1, 2022
AbstractǫŃÚ1ɋC1851 ĸ 1 k L g  Ȟ Ê ě Ȁ 2

Heinrich Müller 1A), ROH3=.

Rhodopsins are photoreceptive membrane proteinsDÓusing retinal chromophore. Two large families ofƳ2ǷȈ1uVȍ2ƳȰĚĀrhodopsins, animal and microbial rhodopsins, wereE&.1AD B1 2ƳȰH·1B

known so far, and they are used by diverse animals and.$2ȍȾȍ

microorganisms, respectively. In contrast, Asgard

ƻěȀ2 Franz Christian Boll 1A),șÁE

archaea2ȚțƑȄ.2ɘ°ţ÷E&

arelivingprokaryotes

which

is

being

D.

D.ćñ

1876 ĸ1kLgƽ5)Ǐ2ƳȰ$

considered to be closest to the last common ancestorƹĀ2kx\-D $2ìÉ3U\Jbetween prokaryotes and eukaryotes, and they were notȥ2qthought to use light for their survival. However, weűžȥ-3ȚǬ.>ô4ED $2Ń2ǔǢ1AC

have recently discovered many rhodopsins in their

kx\2ğ¬3 ǷȈ2Ƈ¬ǰȃ21ĚĀ

Hœó

D rhódon. opsis1ǁƁ

D½ǉȈ1ĬĀ

metagenome-assembled genomes. In particular, one ofDȈcrVȰ-D.

crVȰɍÂ1tc A 2Ȥĩ¬-them, schizorhodopsin, is revealed to be a newGCrhodopsin family which functions as a light-driven

D 11-cis ă2elǆȍû.,Ǵê

inward proton pump. In this review, we will introduceD.-çȚĄ1ïâHǖ

the history of rhodopsins and our studies on therhodopsins of Asgard archaea.
1. ÓƳ.ŇƽƳ Ŗ

ɴ*2kx\sjHî<ď2ÓƳ3·DƼĊ1, ǏHƾ,òý1DƳ¬2Ŀ?ȍHșD.-D

E3ǷȈ1ĚĀ

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Journal of Japanese Society for Extremophiles (2022) Vol.21(1)George Wald B2ǔǢ1A),

el·Hŏ1ɘGD ·`Z.,3&Bǟɪ2

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H. salinarum 2čǅƅHƾ&ǔǢ1A),ëĞïâ.ųB.0)&

61)

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crassaɱ2 cDNA1971 ĸ1 Walther Stoeckenius . Diter

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halobium .ô4E,&ɱ2ǰȃȈ2- ǯȍH

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 H+}x.,3&Bkx\ɰxhQBƌŕE,D.șÁE& 44) ľŵ3:'

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1, all-trans ă2elǴê B110)&ɰþ 2ɱ28,40,48)$E·Hïâ
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D¼í Cl!}x-D 38,52):&Ɠȭ2Ȳ·14

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)系の一つとされている。そしてロドプシンの未知のを意味するヘリオロドプシン（HeR）と名付けられ、機能を解明した研究の中で最も大きなインパクト

既存の微生物ロドプシンと同様に、細菌やアーキア、をもたらしたのが、2002 年から 2003 年にかけて、

真核微生物、巨大ウイルスなど極めて広汎な種に存

ドイツの Peter Hegemann らによって報告された緑在することがわかっている。また既存の動物および藻 Chlamydomonas reinhardtii 由来のチャネルロドプ

微生物ロドプシンはいずれも、N 末端と C 末端がそ

シン（ChR）である

41,42)のものと似ているが

32)+。ChR の構造はやはり BR

れぞれ細胞外と細胞内に位置する配向を細胞膜内+で取るが、HeR はそれとは逆の配向を示すという構、ChR が光を吸収すると Na

2+や H 、Ca などの陽イオンを透過するチャネルがタ造的な特徴を持つ（図１）。しかし、HeR は微生物

ンパク質内に形成され、電気化学ポテンシャルにロドプシンのようなイオン輸送を示さず、また従って、これらの陽イオンが受動的に輸送される。

GPCR でもないことから、その分子機能は不明であそして 2005 年から 2006 年にかけて米国の Karl

り、なぜ広汎な微生物が HeR を有するのかは明らDeisseroth や東北大学の八尾寛らによって、動物のかとなっていない。

神経細胞へ ChR を発現し、光を照射すると神経興奮を誘発できることが報告された 6,25)。このロドプ

3. アスガルドアーキアのロドプシン

シンなど光受容タンパク質を、研究対象とする生物

	
  一般に、地球史上において真核生物はアーキアの種の細胞に遺伝学的に導入し、その活動を光で操作

一部から進化したとされているが、特にアスガルドする技術はオプトジェネティクスと名付けられ、現アーキアは現存の種の中で真核生物の祖先となっ在数多くの生命科学研究に利用されている

たアーキアに最も近縁であるとされている 65)。これ

11,12,36,37,47)。一方で、外向き H+ポンプや Na+ポンプ、まで、アスガルドアーキアが光を利用することは知−

内向き Cl ポンプを用いることで、神経細胞を過分られていなかったが、2018 年にチェコの Rohit Ghai

極させ、神経発火を光で阻害する、抑制性のオプトらを中心とする国際グループの研究によって、アスジェネティクスツールとしての利用も行われていガルドアーキアのゲノム中にもロドプシン遺伝子。しかしこれらイオンポンプは一光子の吸があることが見出された 8)。これらのロドプシン遺収で一つのイオンしか輸送しないため、神経操作に

伝子には既存の微生物ロドプシン（一般に Type 1

必要な量のイオンの輸送を達成するには、非常に強ロドプシンと呼ばれる）や HeR に分類されるもの

い光を用いる必要があり、光毒性などの問題が指摘のほかに、進化系統樹上でちょうどそれらの中間にされていた。しかし、2014 年に陰イオンを輸送す

位置する新たなグループが含まれていた（図 3(A)）。

る

9,17,29)

2,62)

、ポ

このグループは英語の split に対応するギリシア語

ンプ型のロドプシンより極めて弱い光で神経抑制の 接 頭 語 の schizo- を 用 い て 、 シ ゾ ロ ド プ シ ン

が可能であることが示された。さらに 2015 年には（schizorhodopsin, SzR）と名付けられたが、その分

米 国 の John L. Spudich ら に よ っ て ク リ プ ト 藻

子機能は不明であった。

るチャネルロドプシンの改変体が報告されGuillardia theta から、野生型のアニオンチャネルロ

	  そこで筆者らが配列情報をもとに、SzR の遺伝子ドプシン（ACR）が発見され、人工的なものよりもを合成して大腸菌を形質転換し、SzR タンパク質をさらに 1000 倍程度弱い光で神経発火を抑えること

異種発現させた。そしてその大腸菌に光を照射した

が示された

16)

。また最近では微生物ロドプシンの C

ところ、細胞外液の pH の上昇が見られた（図 3(B)）

や、グアニリルシク

24)

ラーゼ 51)、ホスホジエステラーゼ 63)などの水溶性

ノ

酵素ドメインが付加した酵素型ロドプシンも見つm-chlorophenylhydrazone (CCCP)を添加したところ、かっており、ChR とは異なった形でのオプトジェネ

光照射に伴う pH 変化が極めて小さくなった。この末端側にヒスチジンキナーゼ

27)

ティクスへの応用が期待されている

39)。さらに細胞膜の H+透過性を向上させるプロト
フ

ォ

ア

で

あ

る

carbonyl

cyanideことは SzR が光を吸収すると細胞内に H+を輸送す。

	  この様に動物ロドプシンは 170 年以上、微生物ロることを示している。

ドプシンは 50 年以上にわたって、研究が行われて

	  しかし、大腸菌の細胞内の H+濃度は細胞外の溶きたが、PR を発見した Béjà と NaR の発見者である媒中より低いため、この実験からは SzR が H+を能

神取、および筆者らによる国際共同研究によって、動的に輸送する内向き H+ポンプであるのか、受動

既存のいずれのタイプにも属さない、第３のロドプ

的に輸送する H+チャネルのかを区別することがでシンファミリーの存在が 2018 年明らかとなった 46)。

きない。そこで細胞内外の電気化学ポテンシャルを

このロドプシンはギリシア語で「太陽のロドプシン」

自由に変化できるパッチクランプ法を用いてホ乳

15

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)þ 3 J^SkJTJACș*)&\bkx\ 24)ɲ(A) ŇƽƳɰType 1ɱkx\.yQkx\ɰHeRɱ A5\bkx\ɰSzRɱ2Ǫǵɘ°(B) ĐȇȐ1ǅǟǆƹE& SzR 1AD·¯Ěǈ0¼í H+ȸȿƠŏƪĞ ɯȍǹ-ǖE&ŵɗ1
,·HưĨ Ǹȍǹ3xjovPJ-D carbonyl cyanide m-chlorophenylhydrazone (CCCP)HÐ&Ĉê2ǴƄHǖ
:&þ1$E%E2 SzR Hǅǟǆƹ"&ĐȇȐȐ¬2{fj2¿ǎHǖJ^SkJTJ2ŝ* Type 1 kɪǰȃ1ǆƹ"& SzR 2LQȸȿHȧ7&.

 ů-F ɠƙÕě}h\ɀȵDɮƓ2Ȉ

x\3Ďí H+}x-D.ǦȀB2ğɠªHÞÐ&ƵŔ->¼í H ȸȿǕȣEɭ1A),ųB1E,D 24) ń), J^S&.B SzR 3·ɬÓă2¼í H+}x-kJTJ3Ďí H+}xăkx\+D.ųB.0)&

E3 XeR 1Ƕ+í H }xăkx\+*Ǌ2¼í H }xăkx\2ǆș-D¼$,ƑȄżǒ2y

Qkx\2ǟɪ2kx\Hƾ, ·16

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)HÇƾ,D ȊóƤ.1J^SkJT

TM6 . TM7 3 Type 1 1ɮǌëŏC 

J2( TJTJ?jJTJ3 Type 1

B SzR  Type 1 . HeR 2!E1įkx\Hŝ&0ů- J^SkJTJ

ŭ

D2HÅ

D23üɟ-)&3Âę2ƌɄH>.1 SzR 2ƴňHȧ

2-> Ÿ>JTJ.ǎƆƽƳ2Ÿǲ»ɂǘ¶ ƒ1Ŗ

1Ƚ>22*-D.ǿBE,DyL

7D.Hȡ;& :!ĐȇȐBǩȖE& SzRdJTJ EBǟɪ2kx\¹,H

2ǟɰSzR1ɱH ɮɃàęɗÎɩŇɔɰHS-AFMɱ

Ź,D BĘƙǈ0ƼĊ1ƈ<J^SHƾ,ȜĦ&.F SzR 3ɓ¬HĿŕkJTJ ĕƙǈ0ƼĊ1ɆÁ ǎƆƽƳ6

.ųB.0)&.ɆÕƍDɜ1 ǟɪ2kx\Hƾ AC

kx\3Âę.10ďɓ¬ƌɄH­D.ǒBE,D 53)

ɓ¬3 Type 1 kx\2->1 BR 0/đɛ·HĴ;1®DA10)&.ƤɘG)&çȄŏţ÷E,D

D

8)JTJŝ*Âę-șBED>2-C ɓ¬
HĿŕ

D HeR .3Đǅ0D>2-)&

4. SzR 2 H+ȸȿ2ÂęRm_ů ɆÕǪǵƏ- SzR 3 Type 1 kx\. HeR

3?3C Type 1 .ëƍ1 N ŽǤ2D³HǰȃĎ

2ɗ1ªǽ

6íDɎíHŝ*.ǖE& $,B1

D ÉȀA5ŃȀ2!E1

ǰȃ-2Âę2ɎíHȧ7&.F

SzR

ÂɪED7 >3¹ƶǣ2vI

Ȣǰ0ƌɄHŅD&= SzR 2ƒµƌɄH X ǹǴ.

ŶƌɄȞƃ1A),ȧ7&.F

D21*,ȗŪ2ȜƯBƉȠHȒ)&SzR4 .ô4E:!JoɐɎÃ2ǌëŏ1*,ș,;D.ɷ

D SzR 2ƌɄH 2.1 Å 2ÂȞȄ-ƜĞž2ȈȮɂyfV^ɰTM1–7ɱ2( N ŽǤÏBžǊ2yfV^ɰTM3ɱ2ɎÃ3
kx\AC HeR 1Ƚ>2-)&D.1ŕ

BR 1Ƌ=,Ȍ©&ɓ¬ƌɄȜƪE

Type 1&ɰFig. 4(A)ɱ18)

ů-

B

ń),

EB2ƌɄ2ɪ©ŏ

SzR 3 Type 1 1ÂɪEDkx\-

þ 4 \bkx\2 X ǹǴŶƌɄ. H +ȸȿǳȴ 18)ɲ(A) \bkx\ɰSzR4ɱ2ɓ¬ƌɄɰɸǰȃȰ³Bș&ƌɄ ɸǰȃȈBķȒ0ůí-ș&ƌɄɱ (B) SzR4 2·ÒȳŃ1ƽDɗ¬2ïâ^{Vjɰĳɱ.·áŌZLViɰéɱ·ÒȳŃ K L .)&ɗ¬-3el3\fvĉĆǴê H+Õ& 13-cis ă.0DɰǸȍǹɱM ɗ¬1,3\fvĉĆǴêB H+ǰȃȰ³6Ȟɞ Dɰɡȍǹɱ (C) SzR4 2 H+ȸȿ1ɑȘ
0Joɐ. ŒĞED H +2ȸȿǳȴ (D) SzR4ɰ~acɱ. BRɰƚȍɱ2ȈȮɂyfV^2Ɏ
íƗȶ SzR4 -3 TM2 A5 TM6 2ǰȃȰ³ BR AC>Ǔ0),DɰǸȍƯǹǑÞɱ17

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)şÓ2čÕHșD.- H+Häã)&ƵŔ2x

D.ǴȨ+BE& ƒ1 SzR 2 H+ȸȿRm_Hȧ7D&=l

jJV`xc2ƽŕHȜƪ
7,21,33)

D.-

 SzR 2Ĉê3 M ɗ¬1,oǜ2r^[Hƾ,Âę2ïâ^{VD

jčÕHƪĞ

D [v$2ƍ0ȱĎïâčÕ3ȜƪE0)& 24) 2\^ƞHƾ,SzR4 2·áŌHȧ7&

SzR4 3·HïâƄă1ǅŏÕ&.

f\vPj.3ș Glu81 xjJV`xc-D$2Ǵ

D.el all-transK

L.£Ȧ.ǐǍ
H BR .Ɨȶ

M 2ɵ*2ïâ^{D.

DA-D

SzR4 2ƌɄ

TM2 A5 TM6 2ǰȃȰ³

Vj2ǅ0Dɗ¬Hǳ,ŪØɹŪǇ ms -ĖƵƋǤ1Ǔ0),D.G)& E1A

Ŕ6.ŗD·ZLVáŌHǖ&ɰþ 4(B)ɱ 

C

2-ƴ1 M ɗ¬3ǯĎĄ1&D ! = 390 nm

rVȰȔɤB0C¼³1ĚĀ

2 H+}x-3xjJV`xcc

SzR4 2 Glu81 3ǰȃȰ³Ȕɤ2

Ƚ1ïâHŝ*.B 2ɗ¬1,D21ħȽ´1ªǽ

el.$EǴê&\ƕĆ.2ɗ1Ŀ

D..0D B RSB B Glu81 62 H+ǝŕED H+Õ\fvĉĆǴêɰ–C=N+H–ɱB2

Ó2ɃĺAC> Glu81 BǰȃȔɤ62 H+ŨÁ2

ɇƨǈ0 H+2Ȟɞȳ),D.GD

ɃĺɃ0D. Glu81 1 H+Ǵê&ƵŔń), SzR 3 M ɗ¬2ƽŕŵ1el\fÂ·ěǈ1ȜƪE0àú-D.ǿBE+

vĉĆɰRSBɱ2 H HǰȃȰ³6.ŨÁ&.

DM ɗ¬2Ƨȕ2ɜ1Ů&0 H+ HǰȃĎ³B
RSB 1¾Ǵê

DĿ-ãCȻ<.ǿBED

5. ĕƱJTJ2 SzR+ RSB 2 H ɇƨǈ1Ȟɞ,ȸȿEDƑƌ3+

 ŸÄ2J^SkJTJ2 SzR 2ǆșB+Ďí H }x? XeR 2¼í H }x1,

ű:-2ɗ1 A$ 100 ǟɪǞĺ2 SzR ƍ0>»ɂ

D>2-D SzR -32ƌɄHȧ7&

JTJ?ɍ2ǰȐ-ǆșE,D $2-.F

BR 0/ȋǈ0ŇƽƳkx\-3Ŗ

3  Ʃ ŏ 2 J  T J -  D Methanoculleus

RSB 2ǰȃĎ³1*2ȅ H+Õ&ɐŏJoɐ

taiwanensis ? Ʃ Ɲ ƚ  B ǆ ș  E & ż Â ɪ 2ĚĀ RSB 2ƓɠȏHĜĞÕ

Methanoculleus į2JTJɰMethanoculleus sp.ɱQ.,2ŀÌHƄ&DRMcL.ǒBE,D SzR Hŝ*.HșÁ& 34)

E:-ɮƩƼ

SzR -3 $EBɐŏJoɐ2ůĒGE RĊ1,3Ďí H }xă2ŇƽƳkx

McLQ TM7 2 Asp184 &'*-D\2;ćñE,C 26,55,59) EB2JT++.Â)& B SzR 2 RSB 2 H 2äDǰȃĎ³62ɢɠĻÎ3ǌħǈ1Ī0C ¼³
62 H+ǝÓHȳCŴ,D.ōGED+

2ƍ0ƴň3ë¼í H }x-D XeR 1>
șBED>2-D ů Űǒ2 H+}x1, RSB BȞɞ& H+3J^r

Uɐ?Wcɐ0/2xjJV`xc.ô4EDɐŏƕĆ1äƨED$, SzR 1,>ëƍ1ɐŏƕĆ H+

2ǝÓ1ɑȘ10D.ǿBED SzR4 2Ĉê
Ɠ1 TM3 2ǰȃȰ³1 Glu81 ĚĀ
2ƌɄBųB.0)&D.$$,2 Glu81 H+

H 2äƨ-0Wc1čǅ&.
F

ȸȿƠŏĝ¹1ƢĒ&.BLMþ 5 ȁƱŏ SzR 2Ʊčŏğɭɲ
85 °C 1DɮƩƼĊǁƁ2 SzRɰMtSzRɰQ]ɱ MsSzRɰɯɱɱ. SzR1ɰɡɱ.2 ƱčŏɇǞ2Ɨȶ SzR1 3 85 °C 2ɮƩ1 250 ÂǞĺB .89¹,2crVȰčŏ D
 MtSzR MsSzR 1*,3ĐɍÂ2crVȰżčŏ2::-D+2ɇǞ1, RSB B2 H 3xjJV`xc-D Glu81 Hǳǁ,ǰȃĎ³6.ŨÁ
ED.ǿBEDɰFig. 4Cɱ 2 RSB BxjJV`xc62 H+2äƨ1*,+H }x2ĈêSzR ¢Ď2ĎíA5¼íȱĎÂ·ƪĞ1A), C=O ¨Ǽ18

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)

アが SzR を持つことは、より多様な機能を持つロド

共通点は同じ内向き H+ポンプという機能を達成す

プシンが高温環境下で用いられていることを示唆るため、分子レベルでの収斂進化が起きたことを示していると考えられる。

唆している。

	  そこで我々は M. taiwanensis と Methanoculleus sp.

	  最後に ATP 合成などの駆動力を与える外向き H+のロドプシン（MtSzR、MsSzR）の熱安定性を、アポンプとは対照的に、SzR が行う内向き H+輸送のスガルドアーキア由来の SzR1 と比較した（図 5）。

生理的意義はいまだ不明であるが、同じく機能が未その結果、界面活性剤で可溶化された SzR1 は 85 °知である HeR と共に、今後難培養性のアスガルド

C の条件下で、ほぼ全てのタンパク質が 250 分以内

アーキアの培養技術や遺伝学的研究手法が確立しに変性してしまったのに対し、MtSzR と MsSzR はていくことで 19)、その役割が明らかになっていくと同条件下においても大部分のタンパク質は変性を期待される。

示さなかった。これは外向き H+ポンプ型以外では初めての耐熱性を持つロドプシンであり、高熱環境

謝辞

下においても SzR の内向き H+ポンプ機能がアーキ	  本稿で紹介したアスガルドアーキアのロドプシ

アの生存に関わっていることが示唆される。MtSzR

ンに関する研究は、筆者が東京大学物性研究所におや MsSzR がどの様にして高い熱安定性を実現していて主宰する研究室のメンバーおよび、名古屋工業

いるか、その分子論的なメカニズムは不明であるが、大学・神取秀樹教授ならびに角田聡特任准教授、細今後 X 線構造解析などを通じて明らかにしていく

島頌子博士、今野雅恵博士、Manish Singh 博士、中ことを計画している。

村良子氏、富田紗穂子氏、イスラエル工科大学・
Oded Béjà 教授、チェコ科学アカデミー・Rohit Ghai

6. 終わりに博士、名古屋大学・内橋貴之教授、生命創成探究セ	  動物と微生物由来の二つのロドプシンは光受容

ンター（ExCELLS）・渡辺大輝博士、東京大学・濡

タンパク質のモデル系として長年にわたり、数多く

木理教授、志甫谷渉助教、樋口晶光氏、その他数多の研究の対象とされてきた。さらに近年のゲノム解

くの共同研究者の方々のご協力によって達成され析技術の発展に伴い、新奇機能を持つ分子の数が爆たものである。これらすべての方々に、この場を借発的に増加し、共通の構造から機能の多様化をもたりて厚く御礼申し上げます。

らす分子メカニズムの解明が急速に進められている。そしてこれらの知見をもとにした新たなコンセ

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23

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)Journal of Japanese Society for Extremophiles (2022) Vol.21(1), 24-38

ORIGINAL PAPER

Koei Hamana1, Hidenori Hayashi1, Takemitsu Furuchi2,
Masaru Niitsu2, Takashi Itoh3, Mitsuo Sakamoto3
and Moriya Ohkuma3

Additional polyamine analysis of newly validated archaeal halophiles,
methanogens, thermophiles, acidothermophiles, and mesophilicammonia-oxidizers for a chemotaxonomy in the phyla Euryarchaeota,
Crenarchaeota, and Thaumarchaeota-Polyamine catalogues of bacterial and archaeal extremophiles- (XI)

1

Faculty of Engineering, Maebashi Institute of Technology, 460-1 Kamisadori-machi, Maebashi, Gunma 371-0816,

Japan.
2

Faculty of Pharmacy and Pharmaceutical Sciences, Josai University, 1-1 Keyakidai, Sakado, Saitama 350-0295, Japan.

3

Japan Collection of Microorganisms (JCM), BioResource Research Center, RIKEN, 3-1-1 Takanodai, Tsukuba,

Ibaraki 305-0074, Japan.

Corresponding author: Koei Hamana, koeihamana@gmail.com
Phone: +81-27-234-4611, Fax: +81-27-234-4611
Received: Jan. 6, 2022/ Revised: Feb. 14, 2022/ Accepted: Mar. 8, 2022
Abstract

Cellularpolyamines

N4-bis(aminopropyl)spermidine.

acid-extracted

In thaumarchaeotes,

from newly available 31 strains of the class Halobacteria,

mesophilic ammonia-oxidizing Nitrosopumilus zosterae

one strain of the class Methanobacteria, one strain of thecontained putrescine and homospermidine, whereas

class

Methanocococi,

Methanomicrobia,
Thermoplasmata

five

one
and

strainsstrain

three

of
of

strains

of

the

class

thermoacidophilic,sulfur-

the

class

Conexivisphaera calida contained putrescine, spermidine

the

class

and

spermine.

and

Two

iron-reducing

alkanolpolyamines,

Thermococci of the phylum Euryarchaeota, and twoN-(2-hydroxyethyl)-1,3-diaminopropane

strains of the class Thermoprotei of the phylum(N-(3-aminopropyl)aminoethanol)

Crenarchaeota,

and

one

strain

of

the

class

N-(3-hydroxypropyl)-1,3-diaminopropane

Nitrososphaeria

and

one

strain

of

the

and

class

(N-(3-aminopropyl)aminopropanol) were detected inConexivisphaeria of the phylum Thaumarchaeota, were

thermoacidophilic Sulfuracidifex metallicus belonging to

additionally analyzed by HPLC and HPGC-MS.

the class Thermoprotei.

The

New polyamine data were

halophilic

combined into our final archaeal polyamine distributionarchaea Halanaeroarchaeum and Natrarchaeobaculum

catalogue for a chemotaxonomy in the domain Archaea.

anaerobic

sulfur-respirating

extremely-

were poor in agmatine, spermidine, and spermine.
Extremely-thermophilic
Methanofervidicoccus

Keymethanogenic
abyssicontained

words:

haloarchaea,

thermophile, Archaea

spermidine,

spermine, and a quaternary branched penta-amine,24

polyamine,

methanogen,Journal of Japanese Society for Extremophiles (2022) Vol.21(1)Natrarchaeobaculum40-42) were newly analyzed in the

Introduction
Inthedomain(haloarchaea),

Archaea,extremeanaerobic

hyperthermophilesinpresent study.

methanogens,and(thermoacidophiles)

halophiles

combined with the polyamine catalogues summarized byacidothermophiles

additiontoFurthermore their polyamine data are

ourprevious

haloarchaeal

polyamineanalyses4,5,12,13,15,18,26,43), as shown in Table 1.

mesophilicammonia-oxidizers, have been isolated from various

The present final report on archaeal polyamines in ourterrestrial and marine environments, and furthermore,polyamine studies showed the first polyamine analysis of

1,24,35)

Acidophiles andnovel acidothermophilic, sulfur- and iron-reducing

alkaliphiles in haloarchaea, and thermophiles, halophiles

Conexivisphaera calida isolated from an acidic hot

and alkaliphiles in methanogens have been found.

spring in Japan and belonging to the second classfrom human associated samples.

Allarchaeal isolates reported so far belong phylogeneticallyConexivisphaeria of the phylum Thaumarchaeota27).

to only several clades located within three phyla.

novel

They

mesophilicammonia-oxidizingAarchaeon,are classified into the nine classes; Halobacteria,

Nitrosopumilus zosterae33) was available as the third

Methanobacteria,

species of the genus of the first class Nitrososphaeria of

Methanocococi,

Methanomicrobia,

the phylum Thaumarchaeota.

Methanopyri, Methanonatronarchaeia, Thermoplasmata,

It has been reported thatand Thermococci in the phylum

these ammonia-oxidizing archaea are predominantEuryarchaeota, the class Thermoprotei in the phylum

among ammonia-oxidizing prokaryotes (bacteria andCrenarchaeota, and the two classes; Nitrososphaeria andarchaea) in soils30). Thermophilic, alkaliphilic, andConexivisphaeria in the phylum Thaumarchaeota1,24,35),

halophilic members are distributed within methanogenic

Archaeoglobata

Meanwhile, over 15

archaeal clades, therefore a new extremely-thermophilicarchaeal phyla, such as candidatus Aigarchaeota,

methanogen, Methanofervidicoccus abyssi, belonging toKorarchachaeota, Nanoarchaeota (Nanoarchaeia) andthe class Methanococci of the phylum Euryarchaeota39),

anaerobic methanotrophic (methane-oxidizing) archaea

was analyzed.

as shown in Tables 1 and 2.

Newly analyzed polyamine data on the(ANME), have been left uncultured or cultured but notthree novel archaea and 12 other additional archaeal

released in research communities24,35).

methanogens, thermophiles and acidothermophiles were

By our extensive cellular polyamine analyses ofcombined in a polyamine catalogue of representative

culturable archaea and bacteria, which have been

culturable archaea previously reported6-9,11-13,15-17,20,21,43),

continued

as shown in Table 2.

from

the

early

researchesofarchaea(archaebacteria) in Japan 28), it has been suggested that
cellularpolyamine

distribution

profiles,as

Since our cellular polyamine analyses of many archaeaaandbacteria

suggested

that

major

polyamine

chemotaxonomic marker, are related to their optimum

compositions, as polyamine distribution profiles, were

growth temperature, pH, and salt concentration, in

not varied within their genus level2,14,19), a representative

additiontoclassified

species in each genera is shown as a reference in Tables

locations

2,14,16,19)

Ionic interaction of polycationic

1 and 2; thus discussion on the polyamine distribution

polyamines such as aliphatic linear diamines, triamines,

profiles for chemotaxonomy to phylogenetic diversity

tetra-amines,

their
.

phylogeneticallyandand for adaptation to growth environments withinbranched tetra-amines and penta-amines, to nucleic acids

penta-amines,

andhexa-amines,

culturable validated archaea was added in this our final

and other acidic components in the cells, is important

report.

especially for extremophilic archaeal and bacterial
growth31,34,38,44).

Materials and Methods

Various haloarchaea belong to the class Halobacteriaof the phylum Euryarchaeota.

Archaeal strains were provided by JCM BRC RIKEN,

More than 60 genera are

Japan and NBRC NITE, Japan and cultivated in thecurrently validated and located in one of the three orders;liquid culture media (100-1000ml) designated in the36)

Natrialbales .

JCM Catalogue 25) or the NBRC Catalogue 35), at the

Polyamines of an additional 31 strains belonging to 24

temperature, pH, and NaCl concentration, listed indifferent

Tables 1 and 2.

Halobacterales,
generasulfur-respiratory

Haloferacales
including
genera

and
the

novel

anaerobic,

Halanaeroarchaeum

andThe archaea in the stationary phasewere harvested by centrifugation.

25

The pellets ofJournal of Japanese Society for Extremophiles (2022) Vol.21(1)archaea (0.5-10g) were washed with 0.8% NaCl or 10%

found, effect of culture temperature and pH conditions

NaCl (for extreme halophiles) and then homogenized byfor their polyamine contents are described below.

ultrasonic disruption in equal volumes of cold 1M
15)

perchloric acid (PCA) .

In the order Halobacterales, a low level of putrescine

Polyamine extraction with

0.5M PCA was repeated two times.

(4), spermidine (34), spermine (343), and agmatine

The PCA-extract

(Agm)

was

found

ubiquitously.

Alkaliphilic

was subjected to a Dowex 50WX8 column (1cm I.D. x

Natronoarchaeum species grown at pH 8.8 have high3cm) and then the column was washed with 1M HCl.

agmatine content.

Polyamines were eluted with 6M HCl from the column.

have similar polyamines to those of neutrophiles.

The concentrated polyamines were analyzed by HPLC

was observed that a slight thermophilic growth condition

on a Hitachi L6000 using a column of cation-exchange

at 45-50℃ did not affect polyamine profiles.

The acidophiles grown at pH 4.5-5.0
Itresin, Hitachi 2619F (=Hitachi 2720) (4mm I.D. x

In the order Haloferacales, an alkaliphile, Halopavum50mm) and determined by post-labeled fluorometry on aalkalitolerans, has no significant amount of polyamines.

Hitachi F1050 Fluorescence Spectrophotometer afterA relatively high concentration of spermidine and

3,13)

After

spermine were found in the two species of Halorubrum.

heptafluorobutyrization of the concentrated polyamine

A high agmatine level was sporadically found insamples, HPGC-MS on a JEOL JMS-700, equipped withneutrophilic species within the order.

being

a

heatedcapillarywitho-phthalaldehyde

column

of

Inert

.

Cap

1MS

Alkaliphiles and neutriphiles are distributed in the(polydimethylsiloxane, 0.32mm I.D. x 30m) (GLSciences) was performedofpolyamines

3,13)

.

Cellular concentrations

andorder Natrialbales.

Most alkaliphiles contained a

relatively high level of agmatine and cadaverine (5).

A

alkanolpolyamines

high spermidine content was detected in a Natrialba(polyaminoalkylalcohols) (µmol/g wet weight of cells or

species, however its level did not vary under different pH

nmol/g wet weight of cells) were estimated from theand temperature as shown previously 12,43).

peak height in comparison with standard polyamines andpolyamine level of a Haloterrigena species did not varyalkanolpolyamines in HPLC and HPGC.

under temperature condition between 30-60℃ and under

A low

Numeric codes of polyamines abbreviated as number

pH condition between 6.5-8.5, as shown previously12).

of methylene (CH2) groups between amino (NH2) or

Polyamines of the newly- analyzed seven strains of theimino (NH) groups are used in Tables 1 and 2 of theorder had similarly low levels, as shown in Table 1.

present study.	
 clear phylogenetic correlation of their polyamineA

distribution profiles was not found among the three
Results and Discussion

orders Halobacterales, Haloferacales and Natrialbales.

Results of the additional new polyamine data in the

Halanaeroarchaeum sulfurireducens belonging to thepresent study and additional chemotaxonomic polyamine

orderdistribution profiles are discussed in the five archaeal

sulfurireducens belonging to the order Natrialbales

groups (1)-(5) separately, followed by our previous

inhabiting the same environment of a hypersaline

discussions on archaeal polyamine distributions in thelake40-42 and cultivated in similar anaerobic growthseries reported in this journal
1)

The

class

11-13,15,16)

Halobacteria.

of

Halobacteriales

conditions,thephylumwere

andsimilarly

lowNatrarchaeobaculum

in

the

commonpolyamines spermidine and spermine (Table 1).

Euryarchaeota

In conclusion, haloarchaeal polyamines are putrescine,

Polyamines of 31 strains belonging to 24 genera which

cadaverine, spermidine, spermine and agmatine, itswere newly available, were analyzed in the present study

cellular concentration varied at low levels, and clear

and then the polyamine data were added into thephylogenetic differences on their distribution profiles

polyamine catalogues of 41 representative species of thewere

175 strains previously analyzed, as shown in Table 1.

Furthermore little variation of the profiles was observed

Unavailable strains of eight haloarchaeal genera in theamong slight acidophilic, alkaliphilic, and thermophilicpresent study are shown in the table.

species of haloarchaea.

Although within

not

found

withintheclass

Halobacteria.

It has been shown that some of

haloarchaea grown under 2-5M NaCl, alkaliphiles andthe cellular polyamines leak into culture media or that

acidophiles were isolated but strict thermophile was not

uptake from the polyamine-containing media used for the26

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)culture of haroarchaea, under a high salt growth
condition
2)

4,5,43)

The

classes, were affected not only by their classified

may occur.

classes

location under same growth conditions (temperature, pH

Methanobacteria,

Methanomicrobia,

Methanococci,

Methanopyri

and salt concentration), but also by growth conditions

andwithin same taxon such as class, order, family and genus.

Methanonatronarchaeia of the phylum Euryarchaeota

3) The classes Thermoplasmata, Archaeoglobata andWe have reported the distribution of a quaternary

Thermococci of the phylum Euryarchaeota

4

branched penta-amine, N -bis(aminopropyl)spermidine

In

the

order

Thermoplasmatales

of

the

class

(3(3)(3)4) in addition to a triamine, spermidine (34), andThermoplasmata, a strain (BRGM4) of Ferroplasmaatheacidiphilum distributed in an acid mine drainage,

andGermany36) was additionally analyzed in the present

Methanotorris species of the class Methanoccoci, grownstudy, and contained spermidine (34) as the major

tetra-amine,

spermine

hyperthermophilic
at 80℃

13,15).

(343)

withinMethanocaldococcus

The branched penta-amine was also foundpolyamine, and putrescine (4) and spermine (343) as thein extremely thermophilic Mathanofervidicoccus abyssi

minor polyamine (Table 2).

grown at 70℃, as shown in the present study (Table 2)

study, high levels of homospermidine (44) and spermine

whereasweremoderately/slightly

thermophilic

found

in

the

As shown in the previous
mesophilic,

neutrophilic,

Methanothermococcus grown at 60 ℃

lacked themethanogenic Methanomassiliicoccus luminyensis whichbranched penta-amine within the class.

Linear andwas isolated from human feces, locating in the orderbranched penta-amines have never been detected in the

Methanomassiliicoccales of the class Thermoplasmata11).

hyperthermophilic Methanothermus species (grown at

On cellular polyamine profiles, the unique order

80℃) or the moderately thermophilic Methanothermobacter

Methanomassiliicoccales was distinguished from anotherspecies (grown at 65℃) of the class Methanobacteria, or inorder

the hyperthermophilic Methanopyrus species (grown atmesophilic/slightly-thermophilic

acidophiles,

95 ℃ ) of the class Methanopyri (Table 2).

Cuniculiplasma,

Ferroplasma,

The

Thermoplasmatalesconsisting

Acidoplasma,ofisThermoplasma, Thermogymnomonas and Picrophilus

phylogenetically significant among different classes ofisolated from acidic terrestrial environments (Table 2).

methanogenic archaea in addition to the degree of their

Thermoplasmatales is classified into the five different

thermophily.

families in the order, however the former five areoccurrence

of

the

branched

penta-amine

Within the class Methanomicrobia, homospermidine

cell-wall-deficient genus whereas an S-layer of cell(44)-dominant genus, spermidine (34) and spermine

surface is present as a cell wall in the last genus24).

(343)-dominant genus, and spermidine, homospermidine

Slightly thermophilic Thermogymnomonas acidicola

and spermine-dominant genus were widespread.

Sincegrown at 60 ℃ contained spermine whereas slightly

this class consists of the three orders; Methanomicrobiales

thermophilic Thermoplasma acidophilus grown at 60℃

including nine genera, Methanocellales including one

lacked spermine11), suggesting that Thermogymnomonas

genus and Methanosarcinales including ten genera,(family is unclassified) is phylogenetically distant fromdetailed description of phylogenetic or chemotaxonomic

the other four families.
New members belonging to the class Archaeoglobata

significance among the three polyamine-distribution
types

is

difficult.

Halophilic

were not available in the present study.

Methanohalobium

Only one genus

andArchaeoglobus is validated and it has been shown that

Methanohalophilus species under 2-7% NaCl contained

several species of the genus ubiqitously contained

a low level of spermidine and spermine (Table 2).

sprmidine,

evestigatum

has

grown

been

under

shown

25%

that

NaCl

It

9.0

and

under

13%

NaCl,

Methanonatronarchaeia,

contained

spermidine,(Tableofthenorspermine

N4-bis(aminopropyl)norspermidine

Methanonatronarchaeum thermophilum grown at 45℃,
pH

spermine,

N4-aminopropylnorspermidine

haloalkaliphilic

4

N -bis(aminopropyl)spermidine

class

Quantitative

putrescine,

analysis

of

(333)

(and

(3(3)3)),(3(3)(3)3)(3(3)(3)4)
norspermine

and11,13).

andCellularN4-aminopropylnorspermidine was difficult in thesepolyamine compositions of archaeal methanogens whicharchaea, because separation of 333 and 3(3)3 is difficult

are phylogenetically classified into the five different

in HPLC, and 3(3)(3)3 and 3(3)(3)4 are destroyed toandspermine2).

27

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)produce 3(3)3 during HPGC in the present study.

(N-(3-aminopropyl)aminopropanol)

Occurrence of the two quaternary-branched penta-amines

detected in Sulfuracidifex metallicus JCM 9184, as

is a character of the neutrophilic, extremely thermophilic

shown in Table 2.

genus (Table 2).
found

as

were

Although alkanolpolyamines havebeen found in the thermophilic bacteria Thermus,

In the class Thermococci, N4-bis(aminopropyl)spermidine
was(33OH)amajorpolyamineinMeiothermus and Calidithermus10), this is the first reportmarineon the occurrence of alkanolpolyamine in archaea.

Thermococcus indicus and Thermococcus sp. 5-4, andLinear penta-amines and hexa-amines have beenmarine Pyrococcus sp. PK 501, which have been newly

detected

available in the present study (Table 2).

The ubiquitous

Thermoproteales but not in the orders Acidilobales,

occurrence of the quaternary branched penta-amine,

Sulfolobales and Fervidicoccales, furthermore branchedputrescine (4), spermidine (34), spermine (343), andpolyamines have never been detected in the class.

agmatine (Agm) in other Thermococcus (26 species),

Except for phylogenetic distances within the class,

Pyrococcus (6 species) and Palaeococcus (3 species)

growth temperature (60-100 ℃ ) and pH (1.3-8.0)

species belonging to the order Thermococcales (classconditions might affect polyamine distribution profiles.

Thermococci) has been reported

11-13,15,16)

.

in

the

ordersDesulfurococcales

andAt the culture

Without statistical polyamine analyses, we avoid thetemperatures 65-95 ℃ ,the two terrestrial freshwaterdescription of the effects in detail for their growth

Thermococcus species, T. waiotapuensis and T. zilligii

conditions.

were cultivated in the medium containing 0.25-0.75%(44) in Sulfophobococcus species, and homospermidine

NaCl and other halo-tolerant marine Thermococcus,

and aminopropylhomospermidine (344) in Thermogladius

Pyrococcus and Palaeococcus species were cultivated in

species, are novel within the class (Table 2).

the medium containing 0.70-2.5% NaCl.

5) The classes Nitrososphaeria and Conexivisphaeria

The branchedpenta-amine level decreased under a lower saltof the phylum Thaumarchaeota

concentration or a lower growth temperature in
Thermococcus species, as previously reported

11,16)The major occurrence of homospermidine

Mesophilic.

ammonia-oxidizing

Nitrosopumilus

zosterae isolated from a coast in Japan33)

was analyzed in

Within archaea, sporadic occurrence of canavalmine

the present study.

(434) was specific for the order Thermococcales.

other Nitrosopumilus species contained putrescine (4)

4)

The

class

Thermoprotei

of

the

phylumThis new species as well as twoand homospermidine (44) (Table 2).

Crenarchaeota

Thermoacidophilic,

Hyperthermophiles

andacidothermophiles

sulfur-andiron-reducing

areorganoheterotrophic Conexivisphaera calida located in

distributed in the five orders of the class Thermoprotei

the new class Conexivisphaeria contained spermidine(one class belongs to Crenarchaeota)

36)

and previous(34) and spermine (343) as the major polyamine andpolyamine profiles of the representatives are summarized

diaminopropane (1,3-diaminopropane) (3) and putrescine

in Table 2.

as the minor polyamine.

It has been shown that the occurrence ofnorspermidine(33),

norspermine(333),

andThe high content of spermine,a tetra-amine, found in C. calida grown at 65℃ seemed

caldopentamine (3333) was a chemotaxonomic interest

to be correlated to its slight/moderate thermophily.

among the orders or families in the class.

	 Type strain

On

the

other

hand,

anothermesophilicof Sulfuracidifex metallicus and a strain (N-8) of

ammonia-oxidizing Nitrososphaera species in addition to

Sulfolobus

acidocaldaris

acidothermophilicorderbelongingtotheNitrosopumilus

Sulfolobales

the

class

Nitrososphaeria

speciesbelonging

contained

to

the

class

putrescine

and

11)

Thermoprotei were additionally analyzed in the present

homospermidine as the major polyamine , as shown instudy.

Table 2.

(4),

The two acidothermophiles contained putrescinenorspermidine,

spermidine(34),

tetra-amine

norspermine,

two(32OH)

areremarkably

different

putrescine and homospermidine were found in thealkanolpolyamines,

archaea belonging to the class Nitrososphaeria whereas

N-(2-hydroxyethyl)-1,3-diaminopropane(N-(3-aminopropyl)aminoethanol)

distributions

between the two classes of the phylum, however

spermine (343), and thermospermine (334).
Furthermore,

It is suggested that diamine, triamine and1,3-diaminopropane,andputrescine,spermidine

andspermine were found in the archaeon belonging to theN-(3-hydroxypropyl)-1,3-diaminopropane

28

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)(Ac3(3)(3)4) in Thermococcus kodakarensis.

class Conexivisphaeria.

2-Hydroxyputrescine

Polyamine components and distribution profiles in

Betaproteobacteria

Although studies on enzymatic linear and branchedsyntheses

andcharacterization

reported

and

22,29,32,37),

wediscussedandHydrogenophilalia

of

the

bacterial phylum Proteobacteria2), have never beenofaminopropyltransferases in thermophilic archaea have
been

Hy4)

2-hydroxyspermidine (H-34, Hy34) found in the classes

Archaea and Bacteria
polyamine(H-4,

polyaminefound in archaea.

N-Methylated polyamines such as

methylputrescine

(M4,

Me4),

N4-methylspermidine

5components and modified polyamines in our cellular(M34, Me34) and N -methylspermidine (3(M)4, 3(Me)4)

polyamine catalogues of culturable archaea belonging tofound in algae and plants, have never been detected in

the three phyla.

both archaeal and bacterial organisms.
In bacteria, several alkanolpolyamines were first foundVarieties of polyamine distribution profiles were
found within the three phyla of Archaea but polyamineincomponents specific for the archaea have never been

Calidithermus

found.

alkanolpolyamines N-(2-hydroxyethyl)-1,3-diaminopropane

and

It is suggested that common ancestor of bacteria
archaea

evolutions

had

the

potential

thermophilic

Thermus,species10).

In(N-(3-aminopropyl)aminoethanol)ofMeiothermus

andarchaea,two

the(32OH)

anddecarboxylation activity for basic amino acids such as

N-(3-hydroxypropyl)-1,3-diaminopropane

ornithine, lysine and arginine to produce putrescine(4),(N-(3-aminopropyl)aminopropanol) (33OH) were foundcadaverine (5), and agmatine (Agm), respectively andin Sulfuracidifex metallicus in the present study.

aminopropyl-transfer

the first report on the occurrence of alkanolpolyamine inactivitytothediaminesorThis isarchaea.

guanidinoamine.
Decarboxylation of aminopropionic acid to produce

Author contributions

1,3-diaminopropane (3) has never been found in archaea
whereas 1,3-diaminopropane produced by the polyamine

This study was carried out as the joint research projectoxidase for spermidine (34), and spermine (343), in“Polyamine distribution profiles as a chemotaxonomic

addition to aminopropionic acid decarboxylase activity,

marker in Bacteria and Archaea” between Maebashi

was found in bacteria.

Both archaeal and bacterial

Institute of Technology (Koei Hamana and Hidenori

hyper/extreme thermophiles grown at a high temperature

Hayashi) and JCM BRC RIKEN (Takashi Itoh, Mitsuo

contained mostly long linear penta (and hexa)-amines or

Sakamoto, and Moriya Ohkuma).

branched penta-amines (N4-bis(aminopropyl)norspermidine

and Masaru Niitsu of Josai University participated in the3(3)(3)3, N4-bis(aminopropyl)spermidine 3(3)(3)4 and

HPGC-MS analyses of polyamines.

4

N -aminopropylspermine 3(3)34).

However, withinbacteria, linear penta-amines and branched penta-amines

Acknowledgments

coexist in the bacterial hyper/extreme thermophile,
3,10)

We thank JCM and NBRC for supplying archaeal

The coexistence of linear andstrains and advice for their cultures, and also thank Dr.

branched long polyamines has never been found in

Sorokin, D.Y. (Delft Univ. of Technol., Netherlands) for

archaea.

andthe deposits of Halanaeroarchaeum sulfurireducens

N-bis(aminopropyl)cadaverine (353) found in some

JCM 30661 and Natrarchaeobaculum sulfurireducens

Thermus species.

Takemitsu Furuchi

Aminopenthylnorspermidine

(335)

2)

bacteria have never been archaea.
Inbacteria,

N-acetylated

JCM 30663.

polyamines

1

such

as

8

N -acetylspermidine (Ac34) and N -acetylspermidine(34Ac) were detected in the family Enterobacteriaceaeof the class Gammaproteobacteria of the phylum
Proteobacteria2).

Inarchaea,

we

have

found

1

N -acetylcaldopentamine (Ac3333) in Staphylothermus
hellenicus21) (not shown in Table 2).

Hidese et al.23)

reported the occurrence of an acetylated branchedpenta-amine,

N4-bis(aminopropyl)-N1-acetylspermidine

29

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)30Journal of Japanese Society for Extremophiles (2022) Vol.21(1)31Journal of Japanese Society for Extremophiles (2022) Vol.21(1)32Journal of Japanese Society for Extremophiles (2022) Vol.21(1)33Journal of Japanese Society for Extremophiles (2022) Vol.21(1)34Journal of Japanese Society for Extremophiles (2022) Vol.21(1)35Journal of Japanese Society for Extremophiles (2022) Vol.21(1)References

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38

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)Journal of Japanese Society for Extremophiles (2022) Vol.21(1), 39-71

ORIGINAL PAPER
Koei Hamana1, Hidenori Hayashi1, Takemitsu Furuchi2,
Takeshi Uemura2, Masaru Niitsu2, Takashi Itoh3, Mitsuo
Sakamoto3 and Moriya Ohkuma3

Polyamine analysis of acidophiles, alkaliphiles, halophiles and thermophiles
belonging to the bacterial phyla Actinobacteria, Aquificae, Bacteroidetes,
Cyanobacteria, Desulfobacterota, Firmicutes, Tenericutes, and Thermotogae-Polyamine catalogues of bacterial and archaeal extremophiles- (XII)1

Faculty of Engineering, Maebashi Institute of Technology, 460-1 Kamisadori-machi, Maebashi, Gunma 371-0816,

Japan.
2

Faculty of Pharmacy and Pharmaceutical Sciences, Josai University, 1-1 Keyakidai, Sakado, Saitama 350-0295, Japan.

3

Japan Collection of Microorganisms (JCM), BioResource Research Center, RIKEN, 3-1-1 Takanodai, Tsukuba,

Ibaraki 305-0074, Japan.
Corresponding author: Koei Hamana, koeihamana@gmail.com
Phone: +81-27-234-4611, Fax: +81-27-234-461
Received: Mar. 11, 2022/ Accepted: Jul. 5, 2022
Abstract
inthe

Cellular polyamine distribution catalogues
domainBacteria

have

providedcovalently linked polyamines in 16 new members of thevaluableclass Negativicutes of the phylum Firmicutes was added

information for their chemotaxonomic classifications andin this paper.

for polyamine profiles essential for extremophilic growthpolyamine data of representative species in the taxa:

environments.

Acid-extracted polyamines from 339

genus, family, order and class, we presented here ouradditional bacterial strains belonging to the eight phyla

final polyamine distribution catalogues of the eight major

Actinobacteria (25 strains), Aquificae (21 strains),

bacterial phyla.

By also referring to our previous

Bacteroidetes (107 strains), Cyanobacteria (3 strains),
Key words: Actinobacteria, Aquificae, Bacteroidetes,

Firmicutes (132 strains), Desulfobacterota (38 strains),
Tenericutes (3 strains) and Thermotogae (10 strains),

Cyanobacteria, Desulfobacterota, Firmicutes, polyamine,

including various extremophiles, were newly analyzed

Tenericutes, Thermotogaeby HPLC and HPGC.

Cellular concentrations ofnorspermidine,

spermidine,homospermidine,

Introduction
To characterize cellular polyamine components ofnorspermine,

spermine,

thermospermine,

bacteria,

diaminopropane, putrescine, cadaverine, diaminohexane,

aminopropylhomospermidine,variouslinearandbranched

polyamine

components such as aliphatic diamines, triamines,

canavalmine,homospermine, caldopentamine, homocaldopentamine,

tetra-amines,

thermopentamine, caldohexamine, homocaldohexamine,

acid-extracted from cultured bacterial cells and analyzed

thermohexamine,by high-performance liquid chromatography (HPLC) andhomothermohexamine,high-performance4

N -aminopropylnorspermidine,
N4-aminopropylspermidine,penta-amines,

2-phenylethylamine were analyzed.

are(HPGC)

Through this process,endogenous cellular polyamine distribution profiles

N4-bis(aminopropyl)norspermidine,
N -bis(aminopropyl)spermidine,hexa-amines

chromatography

developed in our laboratory.

N4-aminopropylspermine,

4

gas

and

agamtine,

withinandbacteria

chemotaxonomic

Occurrence of

39

have

already

provided

information8,21,29,56).

valuableWe

haveJournal of Japanese Society for Extremophiles (2022) Vol.21(1)reported previously several polyamine analyses in the(Table 3), Cyanobacteria (Table 4), Desulfobacterota

class, order, family and genus ranks of the six phyla(Table 5), Firmicutes (Table 6), Tenericutes (Table 7)

Actinobacteria

6,51)

Cyanobacteria

10,27,47),

Bacteroidetes,

24,33,34,35,50),

and Thermotogae (Table 8) are included alphabetically in

6,20,36,51),

the present study to survey the effect of their growthFirmicutes

Aquificae14,20,22) and Thermotogae20,22,39) using validated

conditions to their polyamine profiles.

and available strains before 2006, 2008, 2016, 2014,of polyamine profiles and cellular polyamine levels

2014, and 2014, respectively.

In our previous studies ofshould depend on phylogenetic distance, comparisonsextensive analysis of bacterial polyamines, we attemptedamong these extremophiles within allied genera in same

to decipher polyamine distribution profiles useful for

phylum are required.

chemotaxonomic classifications of bacteria and essential

polyamines of new members of negativicutes (the classfor extremophilic growth environments in bacteria.

Negativicutes) of the phylum Firmicutes are shown

In the present report, polyamine analyses of 298 newmembers belonging to the six phyla are shown.

Since major part

Additionally, covalently linked

separately in Table 9.

To

In this study, we referred to the taxonomy database of

propose polyamine catalogues of so far not investigated

NCBI (National Center for Biotechnology Information,

bacterial taxa of different lineages and to describe

US National Library of Medicine, USA) 60) and added thechemotaxonomic

polyaminepolyamine catalogues of the validated species by

profiles in bacteria, we added the cellular polyamine

analyzing the strains available from the domestic culture

profiles of 38 newly validated strains of the new phylum

collections JCM, MCC-NIES and NBRC.

Desulfobacterota,

class

reclassification of previously validated bacteria is

Deltaproteobacteria of the phylum Proteobacteria andcontinuing, emendation of genus name and its taxon

the former phylum Thermodesulfobacteria63) werename are possible in future.

evaluation

where

of

the

cellularformerSince

combined, as the seventh phylum in the present report.
of the phylum Tenericutes have never been analyzed,

Materials and Methods
Bacterial strains were provided by Japan Collection

polyamines of three representative Tenericutes wereMicroorganisms (JCM) of the BioResource Researchanalyzed here, as the eighth phylum in the present report.

Center (BRC), RIKEN, Japan and by BiologicalSince ionic interaction of highly basic polyamines to

Resource Center (NBRC), of the National Institute ofnucleic acids DNA and RNA and other acidic

Technology and Evaluation (NITE), Japan, and bycomponents in bacterial cells is affected by their growth

Microbial Culture Collection, National Institute forenvironments, cellular polyamine profiles would be

Environmental Studies (MCC-NIES).

diversified depending roughly on growth temperature,

cultivated in the liquid culture media (100-1000ml)

Though cellular polyamines of the cell wall-less bacteria

described in JCM Catalogue53), NBRC Catalogue59) orpH, and salt concentration, in addition to their
phylogeneticallyclassified

The strains were

MCC-NIES Catalogue57) at the temperature and pH,

locations21,22,23).

Furthermore, cellular polyamine levels might affect thelisted in Tables 1-9.

The bacteria in the stationary

cellular production of short chain fatty acids and acidic

phase were harvested by centrifugation.

polysaccharides in bacteria.

bacteria (0.5-10g) were washed with 0.8% NaCl and thenThe pellets ofhomogenized by ultrasonic disruption in equal volumes

Although definition of extremophily for bacterial
growth has not been fully decided, psychrophilesof cold 1M perchloric acid (PCA).

(optimum growth at 5-15°C), slight thermophiles

extraction with 0.5M PCA was repeated twice.

(45-50°C), moderate thermophiles (55-65°C), extreme

PCA-extract was subjected to a Dowex 50WX8 column

thermophiles

hyperthermophiles(1cm I.D. x 3cm) and then the column was washed withacidophiles

1M HCl.

(70-80°C),(hyper-thermophiles)(85-100°C),(pH

Polyamine
The

Polyamines were eluted with 6M HCl from

1.5-5.5), alkaliphiles (pH 8-12), and halophiles (optimum

the column.

growth in the presence of 3-20% NaCl), are covered inanalyzed by HPLC on a Hitachi L6000 using a column

the present report.

Newly available extremophilicof cation-exchange resin, Hitachi 2619F (=Hitachi 2720)

members distributing to the eight phyla; Actinobacteria(4mm I.D. x 50mm) and molecular species were(shown in Table 1), Aquificae (Table 2), Bacteroidetes

determined by post-labeled fluorometry on a Hitachi

40

The concentrated polyamines wereJournal of Japanese Society for Extremophiles (2022) Vol.21(1)F1050 Fluorescence Spectrophotometer after being
heatedwitho-phthalaldehyde

9,19)

.

Nitriliruptorales (alkaliphilic Nitriliruptor alkaliphilus)

Afterand the order Euzebyales (Euzebya tangerina) were

heptafluorobutyrization of the concentrated polyamine

observed (Table 1).

samples, HPGC-MS on a JEOL JMS-700, equipped withalkaliphilic and slightly thermophilic (grown at 45-50°C)

aspecies of Actinomyces, Micromonospora, Nocardia andcapillarycolumn

of

Inert

Cap

1MS

(polydimethylsiloxane, 0.32mm I.D. x 30m) (GLStreptomyces

Sciences) was performed.

spermidine

Cellular concentrations ofMany mesophilic, halophilic,contained
andputrescine,spermineatlowcadaverine,
concentration.

polyamines (per wet weight of cells) were roughly

However, their cellular polyamines were judged to be

estimated from the peak height in comparison withincorporated

standard polyamines in HPLC and HPGC
Numeric

codesofaliphatic

9,19)

.

cultivationpolyaminesare28,29)

from
.

the

media

used

for

their

Thermophilic Rubrobacter species(Rubrobacterales),thermophilic

Thermoasporomycesabbreviated as number of methylene (CH2) groups

species (Propionibacteriales), and thermoacidophilic

between amino (NH2) or imino (NH) groups andAcidothermus species (Acidothermales) as well as othercapitalized abbreviations such as Put (putrescine), Cad

mesophiles belonging to the order Bifidobacteriales,(cadaverine), Spd (spermidine), Spm (spermine), Agm

Euzebyales and Coriobacterales (Atopobium, Collinsella,(agmatine), and Pea (2-phenylethylamine) are also used

Coriobacterium, Eggerthella, Olsenella, and Slackia)in the present report.

contained no significant amount of polyamines6,43,51) andsome of them are shown in Table 1.

Thermophily at

Results and Discussion
	  Results of the new polyamine data in the present study

45-60°C, acidothermophily and alkaliphily does notand chemotaxonomic polyamine distribution profiles are

phylum.

seem to affect cellular polyamine levels within thediscussed in the eight bacterial phyla separately, by also
referring to our previous discussions on bacterial
polyamine

distributions

ourseriesof2) Phylum Aquificae (Table 2)

this

Chemolithoautotrophic thermophiles belonging to theTentatively summarized polyamine

phylum Aquificae are widely distributed and often dominate

profiles in thermophilic bacteria within 13 bacterial

in various sulfidic geothermal environments such as

journal15,17,18,20,22).

inphyla have been shown in our previous report 20).

terrestrial hot spring and deep-sea hydrothermal vents. We
have1) Phylum Actinobacteria (Table 1)
The

classes

Coriobacteriia,

Acidimicrobiia,
Nitriliruptoria,

analyzedpolyaminesof21

extreme/moderate-thermophiles belonging to two orders
Actinobacteria,(total three families) of the phylum Aquificae11,14,19,22,48), as

Rubrobacteria,

and

60)

shown Table 2.

Wejamiesonii belonging to the family Aquificaceae4) washave reported the absence of appreciable amount ofpublished and the occurrence of a branched penta-amine,polyamines in many species belonging to the orders

N4-bis(aminopropyl)norspermidine, was found in the

Actinomycetales, Bifidobacteriales, Coriobacteriales andpresent

Rubrobacterales including moderate thermophiles grownsulfide-oxidizing large sausage-shaped bacterium belonging

Thermoleophilia are included in this phylum .

at 60°C12,51).

By our previous polyamine analysis,

Thermoleophilum

speciesbelongingtothestudy.

A new species of Thermocrinis, T.

Thepolyamine

profileofato the family Hydrogenothermaceae, collected from aorder

sulfidic hot spring (at 75°C and pH 6.5) in Nagabusa, Japan

Thermoleophilales of the class Thermoleophilia, grown

and previously analyzed in our report26) was added in Table

at 70°C, contained norspermidine (33), spermidine (34),N4-aminopropylspermidine
N4-bis(aminopropyl)norspermidine
42.

(3(3)4),(3(3)(3)3)39)

N -bis(aminopropyl)spermidine (3(3)(3)4) .

The sulfide-oxidizing extreme-thermophile is closely

related to the genus Sulfurihydrogenibium, however, it isandunculturable and as not been validated61,62).

Analyzed

In theAquificae bacteria contained ubiquitously one (or two) ofpresent study on the 25 new members, the presence ofthe quaternary-branched penta-amines and did not containcadaverine and spermidine in the order Acidimicrobiales

homospermidine and linear penta-amines.

(Aciditerrimonas, Acidimicrobium, and Iamia) and theabsence of significant amount of polyamines in the order3) Phylum Bacteroidetes (Table 3)41

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)By the addition of the 107 newly analyzed members

polyamine(strains), heterogeneous polyamine profiles have been

profilesconsistingof

putrescineandspermidine, or putrescine and homospermidine.

observed within each order or family of the phylum
Bacteroidetes

consisting

Sphingobacteriia,

of

the

Saprospiria,

sixclasses;4) Phylum Cyanobacteria (Table 4)

Chitinophagia,

The cellular polyamines of 140 cyanobacterial strains

Flavobacteriia, Cytophaggia, and Bacteroidia (the six

have been roughly separated into spermidine-dominantorders Sphingobacteriale, Saprospirales, Chitinophgales,

types and homospermidine-dominant types10,27,47).

Flavobacterales, Cytophagales, and Bacteroidales), as

additionshown in Table 3.

cyanobacteria,

The present polyamine data intomarineandterrestrial

slightly/moderatelyInfreshwater
thermophilic,

addition to some previous polyamine data6,24,33,34,35,37,49,50)

halophilic and thermoalkaliphilic species are distributed

are summarized and are shown in Table 3.

in oxygen-producing photosynthetic bacteria belonging

In

Sphingobacteriales, analyzed Sphingobacterium species

to the phylum Cyanobacteria, as shown in Table 4.

contained homospermidine exclusively as a polyamine.

Isolation sources are also shown in Table 4.

In Saprospirales, polyamine profiles varied among theshown that thermophilic Thermosynechococcus species

seven strains.

In Chitinophaga, homospermidine wasisolated from terrestrial hot spring contained low levels

major in Chitinophaga, Taibaiella, Niabella speciesof putrescine, spermidine, homospermidine and spermine.

analyzed.

Unusual polyamine profile was not found in the

In Flavobacteriales and Cytophagales,

spermidine-dominant type, homospermidine-dominant

halophilic cyanobacteria.

type and spermidine plus homospermidine-dominant
type were found as a triamine type.
speciesandsome

The two additional ‘spirulina’ samples analyzed in the

Flammeovirga

Hymenobacter

species

It has beenpresent study showed similar polyamine pattern found inofprevious ‘spirulina’ samples (Table 4).

Although

Cytophagales did not contain significant amount ofthermospermine (334) is usually detected together with

spermidine

Threenorspermine (333) and spermine (343) in most otherwithinbacteria, thermospermine was found as a major

Some of the spermidine-dominantpolyamine not by accompanying norspermine orandtriamine-dominant
Bacteroidales.

homospermidine.
types

were

species contained spermine.

also

found

The sporadic detection

sperminecadaverine, agmatine and 2-phenylethylamine is not
significant

for

their

chemotaxonomy.

species)24)

Limnospiraplatensis

This novel polyamine type seems to berelated to its haloalkaliphilic (pH 9.2, 1.7% NaHCO3)

Similargrowth condition.
Although most cyanobacteria contain chlorophyll a as a

described in detail in the previous studies on thepolyamine analyses of 41 species

haloalkaliphilic(‘spirulina’) .

discussion on the polyamine distributions has been
50)

inand 71 genera (105major pigment, chlorophyll b-containing Prochlorococcus

of this phylum.

species and chlorophyll d-containing Acaryochloris locate

The occurrence of a pentamine, thermopentaminein the order Synechococcales.

Acaryochloris tomasi,(3343), in addition to spermine (343) has been reported

which is newly analyzed in the present study, commonly

in moderately thermophilic Acetomicrobium flavidum

contained putrescine and spermidine as the major

6)grown at 60°C .

Crenotalea thermophilea grown atpolyamine in the cultures No. 1 and No. 2 (Table 4).

55°C contained the tetra-amines, spermine (343),

Homospermidine and agmatine were also detected in theaminopropylhomospermidine (344) and homospermine

culture No. 2.

(444) in addition to spermidine (34) and homospermidine

marina MBIC 11017 contained spermidine and agmatine,(44) (Table 3).

andHowever Luteonella thermophile andSphingobacterium thermophilum grown at 45°C, andIn the previous studies, Acaryochloris

Acaryochloris

marinadiaminopropane,

NIES

putrescine,

10,27).

2412

contained

spermidine

Althoughdistribution

andofSchleiferia

thermophileandThermaurantimonas

homospermidine

aggregans

grown

50°C

did

homospermidine is a chemotaxomomic marker within the

at

not

containaminopropylhomospermidine,homospermine,thermopentamine (Table 3).

Haloalkaliphiles andphylumandCyanobacteria,

polyamine

profilesareheterogeneous within the cultures, strains or species of thetiny single-celled marine cyanobacteria, Acaryochloris.

psychrophiles in this phylum showed the common

42Journal of Japanese Society for Extremophiles (2022) Vol.21(1)Related bacterial strain belonging to the newly
validated phylum Melainabacteria

60)

palaeochoriense did not contain polyamines whereas

was not available inspermidine and spermine were found in slightly

the present study (Table 4).

thermophilic

Dissulfurimicrobiumhydrothermale.

Syntrophus aciditrophicus contained homospermidine
5) Phylum Desulfobacterota (Table 5)

and spermine.

The class Deltaproteobacteria was formerly described as

Desulfacinum infernum of the class Syntrophobacteria

a class of the phylum Proteobacteria until 2005 and thecontained spermidine and spermine.

members of the class were phylogenetically separated into
the

phyla

Desulfobacterota,
63).

Bdellovibrionota

We

Myxoccota,

have

analyzed

In the class Thermodesulfobacteria, alkaliphilic,andextremely-thermophilic Caldimicrobium thiodismutans54),

cellularwhich was newly analyzed in the present study,

polyamines of 24 deltaproteobacteria belonging to the new
20,23,41,44).

phylum Desulfobacterota

Thermophilic Desulfosoma profundi andcontained

The former phylum

linearcaldopentamine

homocaldopentamine,

and

and

branched

Thermodesulfobacteria in which polyamines of fourN4-bis(aminopropyl)spermidine, as well as extremely

thermophilic species were analyzed previously12,20,48) was

thermophilic Thermodesulfobacterium species (Table 5).

placed as a class, Thermodesulfobacteria, in the new

The two linear penta-amines were not detected in the two

phylum Desulfobacterota consisting 14 classes.

thermophilic Thermodesulfatator species.

Since

alkaliphiles, halophiles, thermophiles and psychrophiles are
distributed within the new large sulfate-reducing phylum6) Phylum Firmicutes (Table 6)

Desulfobacterota, polyamine analyses of 38 newly available

We analyzed 132 firmicutes in the present study andspecies of the phylum were combined into the previous

report their polyamines separately in the following

polyamine catalogues, as shown in Table 5.

classes (orders).

In

the

class

Desulfuromonadida,

thermophiles

A polyamine catalogue is presented inTable 6.

contained spermine (343) as a major polyamine.
Spermine was not found in psychrophiles.
In

the

class

Class Clostridi

Desulfovibrionia,(Order Clostridiales)

N4-bis(aminopropyl)spermidine (3(3)(3)4) in addition to

Various extremophilic bacteria are distributed withinspermine were found in thermophilic Desulfothermus
species

grown

at

60°C.

However,

the

the

genus

order.

Acetomicrobium

faecalehasreclassified as Caldicoprobacter faecalis

3)

been

and wasDesulfothermus had been located in a class of the former

analyzed here.

phylumProteobacteria.

Caldicoprobacter species belonging to the first family

N -Bis(aminopropyl)spermidine, a quarternary branched

Caldicoprobacteracese contained spermidine, spermine

penta-amine,and N4-bis(aminopropyl)spermidine (Table 6).

4

has

been

Thermodesulfatator,
Caldimicrobium

inthermophilic

Thermosulfobacterium,

speciesbelonging

Thermodesulfobacteria
Desulfobacterota

found

of

(Table

this

5).

to

the

new

The three extremely thermophilicfamilies

Clostridiaceae,

Toward

and

four

class

Lachnospiraceae and Ruminococcaceae, polyamines
were analyzed (Table 6).

phylumthe major polyamine.

Fundidesulfovibrio,

Thermophiles of the four families

werefamily Desulfovibrionaceae, which have been analyzed

Youngibacter in which putrescine was major andpreviously

spermine was minor, was unique.

in

the

homospermidine (44).

present

study,

contained

in

They contained spermidine as

Humidesulfovibrio, and Megadesulfobivrio species of the
and

rich

Eubacteriaceae,

Petpococcaceae,

Newly validated Desulfovibrio

spermine.

all

Polyamine

profileofIn the family

thermophilic

speciesofDesulfotomaculum and some mesophilic species ofbiadhensis contained homospermidine.
In the class Desulfobacteria, two Desulfosarcina species

Desufosporosinus as well as previously analyzed

contained homospermidine, whereas spermidine was the major

species6,51) contained a significant amount of spermine inpolyamine in the other seven genera analyzed (Table 5).

addition

The

four

speciesbelonging

to

the

to

spermidine.

Acetoanaerobiumclasses

speciesInofmesophilicthefamilyDesulfobulbia and Desulfarculia contained spermidine as

Peptostreptococcaceae, an alkaliphilic species was richthein spermine.

majorpolyamine.

Thermophilic

Defrrisoma

43

Although new member belonging to theJournal of Japanese Society for Extremophiles (2022) Vol.21(1)family XVII Incertae Sedis of the order Clostridiales wasof new extremophilic members of Bacillales were added

not available in the present study, polyamines ofin the present study.

acidophilic Sulfobacillus containing spermidine, spermine

In the family Bacillaceae, alkaliphilic Alkalibacillus

and agmatine and thermophilic (or alkalithermophilic)

species contained low level of spermidine and spermine

Thermaerobacter species containing linear penta-amines(Table 6).

and hexa-amines, are listed in Table 6.

and thermophilic Aeribacillus species grown at 60°C

(Order Halanaerobales)

contained spermidine and spermine (Table 6).

Another alkaliphilic Amphibacillus species
AceticLow level of putrescine, spermidine and spermine

acid-producing Bacillus species, B. coagulans, B.

were detected in the halophilic bacteria belonging to theacidiproaceus and B. badius, and a novel halophile,families Halanaelobiaceae and Halobacteroidaceae

Salisedimibacterium halotolerance contained spermidine

including new species analyzed in the present study

and spermine (Table 6).

(Table 1) and previously analyzed species

6,51)

.

Since

It has been reported previouslythat linear penta-amine(s) and/or hexa-amine(s) are

they have not been grown on polyamine-free synthetic

distributed

sporadicallywithinthermophilic

media, net polyamine synthetic ability is not clear.

Anoxybacillus,

Bacillus,

Perobacillus,

Geobacillus,

However, the trace or minor polyamine components

Hydrogenobacillus,

found in the most species may be incorporated from the

Caldalkalibacillus species grown at 60-78°C.

complex media containing polyamines.

occurrence(Order Thermoanaerobacterales)

N4-aminopropylspermine (3(3)43), was found in some ofofaSaccharococcus
tertiarybranched

andThe

penta-amine,

the thermophilic species.

Although new members of the order were not

2-Phenylethylamine, an aromatic amine, which has

available, polyamines of Caldicellulosiruptor species
The extreme

been found in alkaliphilic B. cohnii and Bacillus sp. AO1,

thermophiles contained various linear and branched

was not detected in the other firmicutes including

tetra-amines and penta-amines.

alkaliphiles, which was analyzed in the previous andpreviously analyzed are listed in Table 6.

present studies (Table 6).
In the family Paenibacillaceae, a branched tetra-amine

Class Bacilli

and a branched penta-amine have been detected in(Order Lactobacillales)

alkalithermophilic Thermobacillus composti grown at

The polyamine analysis of lactic acid bacteria
belonging

to

the

five

families

55°C.

Lactobacillaceae

Branched tetra-amine, branched penta-amine,(Lactobacillus and Pediococcus), Leuconostococeae

linear penta-amine and linear hexa-amine were not found(Leuconostoc), Streptococcuceae (Streptococcus andin the thermophiles belonging to the families Bacillaceae

Lactococcus),and Paenibacillaceae newly analyzed in the present

Enterococcaceae

Tetragenococcus)

and(Enterococcus

andstudy, as shown in Table 6.

Carnobacteriaceae(Carnobacterium) has been reported5,43,51).

CellularLong polyamines were not found in the thermophiles

concentration of spermidine and spermine in the lacticofacid bacteria varied by culture medium composition,

acidothermophilic Alicyclobacillus species grown at

however, appreciable amount of the polyamines was not

60°C or Planococcaceae containing Ureibacillus species

detected in Pediococcus and Tetragenococcus grown ingrown at 60°C, as shown in Table 6.

5)polyamine-free media .

thefamilies

Alicyclobacillaceae

containing

Thermophilic species grown at 50-60°C, belonging toAlcalibacterium species did not contain cellular

the family Thermoactinomycetaceae contained spermine

polymines (Table 6).

New Carnobacterium species ofas a major polyamine in addition to spermidine6),

Carnobacteriaceae

andfurthermore Geothermomicrobium terrae grown at 55°C,

Oenococcus

speciesofLactobacillaceae contained appreciable amount ofanalyzedinthespermidine.

thermopentamine (Table 6).

(Order Bacillales)

contained only spermidine (Table 6).

	  Cellular polyamines of thermophilic, acidophilic,
Class Tissierellia

alkaliphilic and halophilic bacilli distributed in six
families have been analyzed

6,51)

and cellular polyamines(Order Thissierellales)

44

present

study,

contained

Mesophiles in the family

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)Moderately

thermophilicorders Petrotogales, Kosmotogales, Thermotogales, andTepidimicrobium

xylanilyticum contained spermidine and spermine and the

Mesoaciditogales52,60).

We have analyzed polyamines

occurrence

of

of

of

spermine

18)

thermophily .
amiovorans,

may

be

Mesophilic
Sedimentibacter

relatedtoits

Dethiosulfatibacteracidominovoransthethermophiles

thisphylum12,20,22,38,39,48).

Furthermore, polyamine distribution profiles of 10 newandspecies available in this study were added in Table 7.

Tissierella pigra contained only spermidine (Table 6).

Slightly

thermophilic

Athalassotoga

saccharophila

which is classified in a new order Mesoaciditogales
grown at 55°C52), lacked penta-amines, as well as other

Class/Order OPB 54

species grown at 40-60°C.

Hydrogenispora ethanolica located in the unvalidated
55)

Moderately/extremely

Class/Order OPB 54 of the phylum Firmicutes ,thermophilic species grown at 65-85°C, contained somecontained only spermidine (Table 6).

linear penta-amines and hexa-amines.

7) Phylum Tenericutes (Table 7)

9)

Cellularpolyamines

of

three

strainsastheValidatedmembersofpolyamines

of

the

class

(Table 9)This is the first report on Tenericutespolyamines.

linkedNegativicutes belonging to the phylum Firmicutes

representative Tenericutes were analyzed in the present
study.

Covalently

We have reported the occurrence of the polyamines

the phylum

covalently linked to the alpha-carboxy group of the

Tenericutes are mesophilic wall-less bacteria that do not

D-glutamic acid residue of cell-wall peptidoglycan in thesynthesize precursors of peptidoglycan and are thoughtmesophilic,

neutrophilic,

to emerge through a series of repeated genome

Acetonema,

reductions

Anaerovibrio,fromdistant

phylum Firmicutes.

ancestryintheanaerobic

18

Acidaminococcus,
Dialister,genera;Anaeromusa,
Dendrosporobacter

The phylum consists of the class

Megasphaera, Pectinatus, Propionispira, Selenomonas,

Mollicutes containing the five orders Acholeplasmatales,

Sporomusa, Vellonella, and Zymophilus in 19997) andAnaeroplasmatales,

200242), and Desulfosporomusa, Megamonas, Mituokella,

Entomoplasmatales,

Haloplasmatales, and Mycoplasmatales60).

Propionispora, and Sporolituus in 201225), of the class

In thepresent study, Acholeplasma laidlawii was shown to

Negativicutes (formerly the subbranch Sporomusa).

contain low levels of putrescine, spermidine, spermine
and agmatine.
mesophilic

Negativicutes locates in the phylum Firmicutes, a

It is rare that spermine was detected inbacteria.

However,incorporation

Gram-positive taxon, and lacks murein lipoprotein whichofmaintains the structural integrity of the outer membrane

spermine from the culture medium containing spermineofis possible.

Negativicutes

Mycoplasmoides pneumonia contained

spermidine and agmatine (Table 7).

Halophilic

theenvelopof

typicalconsistofAcidaminococcales(family

Haloplasma contractile representing a new order

Selenomonadales(families

Haloplasmatales of the class Mollicutes of this phylum

Sporomusaceae),

andhas been reported

1,2)

.

60)

The novel Tenericutes grown

Veillonellaceae) .

Gram-negative

taxa.

the

ordersthree

Acidaminococcaceae),
Selenomonadaceae
Veillonellales

and(family

They were isolated mainly fromunder 12% NaCl contained a low level of spermidine andneutrophilic mammalian gastrointestin and oral cavity.

agmatine, as shown in Table 7.

Veillonella species such as V. atipica are known to

It is suggested that themesophilic wall-less bacteria belonging to the phylum

produce propionic acid from lactic acid.

The short

Tenericutes contain a low level of putrescine, spermidine

chain fatty acids might interact with basic polyamines inand spermine.

the cell.
In the present final report of our studies on negativites,8) Phylum Thermotogae (Table 8)
Membersofthephylum

16 strains including the newly validated genera
Thermotogae

arePhascolarctobacterium, Methylomusa and Lucifera andwell-characterized by their unique outer sheath-likenew species of Anaerovibrio, Selenononas, Pectinatus,

structure called ‘toga’ and shows variation in their

Propionispira,degrees of thermophily

52,60).

The thermophilic phylum

Sporomusa,

Desulfosporomusa,

Megasphaera, and Veillonella were analyzed as shown

Thermotogae (class Thermotogae) consists of the fourin Table 9.

45

Some reanalyzed polyamine data and citedJournal of Japanese Society for Extremophiles (2022) Vol.21(1)polyamine data from related species were also shown in

and/or 343.

the table.

Actinobacteria contained 34 and 343.

Although distribution profiles of their

The most species of the phylum
The absence ofcovalently linked polyamine levels did not directly

polyamines was found in mesophilic Conexibacter,correspond to their intracellular polyamine pools and toalkaliphilic

their growth conditions, distribution of covalently linked

Acidothermus.

polyamines within the class has become conclusive bycontained 3(3)(3)3 and 3(3)(3)4.

the present study.

phylumNitriliruptor,

andacidothermophilic

Thermophilic

Thermoleophilum

Cyanobacteria,

In the photosynthetic

34-dominant

speciesand44-dominan species were distributed, and 343 and 334
Universality and peculiarity of polyamine profiles

werewithin bacteria

haloalkaliphiles.

We have proposed universalities and peculiarities byselectively

distributed

in

thermophiles

andThe triamine 44 has never been

distributed as a major polyamine in the bacterial

the distributions of polyamines within (or among)

thermophiles.

bacterial taxonomic ranks and various extremophiles.
We summarize 8 bacterial phyla reported here.

Polyamine analyses of other bacterial phyla

Polyamines 33, 34, 3(3)(3)3 and 3(3)(3)4 were

In the all phyla of the kingdom (domain) Bacteria,

distributed in an order, Aquificales, and 34 and 3(3)(3)4

consisting 37 phyla (including eight phyla in the present

were distributed in another order, Desulfurobacteriales,

report) validated in 2022, polyamine profiles of the two

within the phylogenetically early-branched thermophilicminorphylum Aquificae.
334

were

early-branched

distributed

thermophilic

phylumwithin12,20)

Dictyoglomi

Polyamines 33, 34, 333, 343, andubiquitously

thermophilic

phylumthe

Caldiserica17,20)

andand the radio-resistant/thermophilic

Deinococcus-Thermus9,15,19,40)

elsewhere.

Thermotogae,

phyla

are

shown

The extensive polyamine analyses of thefurthermore 3333, 3343, 3334, 33333, 33433, 33343, andphyla Acidobacteria, Armatimonadetes, Atribacterota,

33334 were found in the extreme thermophilic species inBalneolaeota, Chlorobi, Chloroflexi, Chrysiogenetes,

the

Petrotogales,

Kosmotogales,

Deferribacteres,

Thermotogales and Mesoaciditogales.

In the phylum

Fusobacteria,

fourordersElusimicrobia,

Fibrobacteres,Gemmatimonadetes,

Ignavibacteriae,

Bacteroidetes consisting the six classes Sphingobacteriia,

Kiritimatiellaeota,

Saprospiria, Chitinophagia, Flavobacteriia, Cytophagia

Planctomycetes,

and Bacteroidia, 34 exclusive, 44 exclusive, and 34 plus

Synergistetes,

44 were class-specifically distributed, and 343 wasreported51,16).

distributed sporadically in thermophilic species.

Lentisphaerae,

Nitrospirae,

Rhodothermaeota,

Spirochaetes,

andVerrucomicrobia

have

been

We have reported the polyamine analyses of the

In thesulfate-reducing phylum Dedulfobacterota, 34 exclusive

former phylum Proteobacteria8,29,30,45).

or

Polyamine

howeveranalyses of new members locating in the emended

thermophilic Desulfothermus species of the order

phylum Proteobacteria (the classes Acidithiobacillia,

Desulfovibrionales contained 343 and 3(3)(3)4, and theAlphaproteobacteria,

thermophilic order Thermodesulfobacteriales contained

Gammaproteobacteria, and Zetaproteobacteria) and the33, 34, 333, 343, 334, 3333, 3334, and 3(3)(3)4.

44

exclusive

were

major

profiles,

Betaproteobacteria,

In thetwo new phyla Bdellovibrionota and Myxococcota

large phylum Firmicutes, alkaliphilic species lacked

belonging to the former class Deltaproteobacteria (thepolyamines, and 34 and 343 were distributed in the most

new phylum Desulfobacterota is described in this report),

mesophilic orders.

and the new phylum Epsilonbacteraeota (including the newly

Polyamine 3(3)(3)4 was found inthermophilic Caldicoprobacter, and 333, 343, 334,

proposed but non-validated phylum Campylobacterota)

penta-amines and hexa-amines were distributed incorresponding to the former class Epsilonproteobacteria

alkalithermophilicof the phylum Proteobacteria 60,63) are in progress as ourThermaerobacter,and3(3)34,

final bacterial polyamine analysis.

3(3)(3)3 and 3(3)(3)4 were distributed in thermophilic
Caldicellulosiruptor.

Polyamines 4, 5, and/or 34 arelinked covalently to cell wall in the class Negativicutesof the Firmicutes.

On the other hand, the cell wall-lessspecies of the phylum Tenericutes contained 4, 34,

46

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)Table 1.

Cellular concentrations of polyamines in the phylum Actinobacteria

47

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)48

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)Table 3.

Cellular concentrations of polyamines in the phylum Bacteroidetes

49

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)50Journal of Japanese Society for Extremophiles (2022) Vol.21(1)51Journal of Japanese Society for Extremophiles (2022) Vol.21(1)52Journal of Japanese Society for Extremophiles (2022) Vol.21(1)53Journal of Japanese Society for Extremophiles (2022) Vol.21(1)54Journal of Japanese Society for Extremophiles (2022) Vol.21(1)55Journal of Japanese Society for Extremophiles (2022) Vol.21(1)56Journal of Japanese Society for Extremophiles (2022) Vol.21(1)57Journal of Japanese Society for Extremophiles (2022) Vol.21(1)58Journal of Japanese Society for Extremophiles (2022) Vol.21(1)59Journal of Japanese Society for Extremophiles (2022) Vol.21(1)60Journal of Japanese Society for Extremophiles (2022) Vol.21(1)61Journal of Japanese Society for Extremophiles (2022) Vol.21(1)62Journal of Japanese Society for Extremophiles (2022) Vol.21(1)63Journal of Japanese Society for Extremophiles (2022) Vol.21(1)64Journal of Japanese Society for Extremophiles (2022) Vol.21(1)65Journal of Japanese Society for Extremophiles (2022) Vol.21(1)66Journal of Japanese Society for Extremophiles (2022) Vol.21(1)haloanaerobes
withinGram-positive
eubacteria: Distribution of spermine andagmatine in thermophiles and halophiles.
Microbiol. Cult. Coll. 15: 9-28.
7) Hamana, K. 1999. Distribution of cellwall-linked
polyamines
withintheGram-negative anaerobes of the subbranch
Sporomusa belonging phylogenetically toGram-positive taxa. Microbios 100: 145-157.
8) Hamana, K. 2002. Polyamine distribution
pattern and chemotaxonomy of bacteria.
Microbiol. Cult. Coll. 18: 17-43 (in Japanese).
9) Hamana, K., Furuchi, T., Hayashi, H., Itoh, T.,
Ohkuma, M. and Niitsu, M. 2016. Occurrence
of
two
novel
linearpenta-amines,
pyropentamine and homopyropentamine, in
extremely thermophilic Thermus composti. J.
Gen. Appl. Microbiol. 62: 334-339.
10) Hamana, K., Furuchi, T., Hayashi, H. andNiitsu, M. 2016. Additional analysis of
cyanobacterial polyamines –Distributions ofspermidine, homospermidine, spermine, andthermospermine
withinthephylumCyanobacteria–. Microb. Resour. Syst. 32:
197-186.
11) Hamana, K., Hamana, H. and Itoh, T. 1995.
Polyaminesinthehydrogen-oxidizing
eubacteria
Hydrogenobacter,
Calderobacterium and Hydrogenovibrio andsulfur-redusing archaebacteria Stygiolobus andDesulfurolobus. Microbios 81: 223-229.
12) Hamana, K., Hamana, H., Niitsu, M.,
Samejima, K. and Itoh, T. 1996. Distributionof long linear and branched polyamines inthermophilic eubacteria and hyperthermophilicarchaebacteria. Microbios 85: 19-33.
13) Hamana, K., Hamana, H., Niitsu, M.,
Samejima, K., Sakane, T. and Yokota, A. 1993.
Tertiary and quaternary branched polyamines
distributed in thermophilic Saccharococcus
and Bacillus. Microbios 75: 23-32.
14) Hamana, K., Hamana, H., Shinozawa, T.,
Niitsu, M., Samejima, K. and Itoh, T. 1999.
Polyamines of the thermophilic eubacteria
belongingtothegeneraAquifex,
Thermodesulfobacterium,
ThermusandMeiothermus,
andthethermophilic

Author contributions
This study was carried out as the joint research project“Polyamine distribution profiles as a chemotaxonomic
marker in Bacteria and Archaea” between Maebashi
Institute of Technology (Koei Hamana and Hidenori
Hayashi)andJCM, RIKEN

BioResource

Center(Takashi Itoh, Mitsuo Sakamoto and Moriya Ohkuma).
Takemitsu Furuchi, Takeshi Uemura and Masaru Niitsu
of Josai University participated in the HPGC and
HPGC-MS analyses of polyamines.
Acknowledgments
We thank Dr. Koji Mori and Dr. Yoshihito Uchino of
NBRC for their advice for the culture of NBRC bacterial
strains.

We also thank MCC-NIES for the supply of

cyanobacterial strains.
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71

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5,000 ´ʯɩ·ēǟ*<ȫ92 65 Ʒu+-'&Ƶì0ǣɕ.9;ńʢ 2,000 ´+!<+*<Àȫ/āá0ųħƁì/Ȧäŕȫ/āá0ņ.9<ŕũɒ?śɅ+!<ʰʯƯʰȉʶƠ Ɲ0ǹȅǪɀÖ2ɘǑȋ/ʌ§ưʑǞăǟǘēɑʯȶäʽJournal of Japanese Society
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73

Journal of Japanese Society for Extremophiles (2022) Vol.21(1)3.	
  会長、副会長の任期は２年とし重任を妨げない。
（評議員会）
4. 評議員の任期は２年とし重任を妨げないが、原則として連続 2 期を限度とする。但し、会長が指名する者はこの限りではない。	
 第１３条	
  評議員会は会長、副会長、評議員をもって構成する。会長は年１回以上評議員会を召集し、議長となり、会の重要事項を審議し総会に提案する。

5. 幹事長の任期は 2 年とし重任を妨げない。
（総会）
6. 庶務幹事の任期は 2 年とし重任を妨げないが、連続 2 期を限度とする。
7.	
  年会幹事の任期は 2 年とし 1 期限りとする。

第１４条	
  本会は原則として年１回総会を開き、会務を協議し議決する。
総会は会長が召集する。

8. 庶務幹事、年会幹事以外の幹事の任期は 2 年とし重任を妨げない。

第１５条	
  総会の議決は出席会員の過半数の賛成をもって行う。

（名誉会長）（会計年度）

第１１条	
 

第１６条	
  本会の事業年度は 4 月 1 日に始まり、翌年 3 月 31 日に終る。

本会には名誉会長をおくことができる。

2. 名誉会長は、本会に対し大局的な見地による適切な助言を与える。

（付則）

3. 名誉会長は、会長として学会の運営及び発展に特別の功績があった者で、幹事会による推薦、評議員
会を経て、総会の承認を受ける。

第１７条 	
  本会則の施行および本会の運営についての細則は総会の議決を経て別に定める。
第１８条 	
  本会則の変更は総会の議決を経て行う。

（委員会の設置と委員の任期）
第１９条	
  本会則は、1999 年 10 月 19 日より施行す
る。

第１２条	
  本会に学会誌編集委員会、研究奨励賞選
考委員会、若手シンポジウム委員会を置く。

2001 年 11 月 29 日改定
2002 年 12 月 1 日改定
2003 年 12 月 1 日改定
2005 年 11 月 7 日改定
2007 年 12 月 1 日改定
2010 年 1 月 1 日改定
2012 年 12 月 1 日改定
2013 年 10 月 26 日改定
2017 年 10 月 16 日改定
2018 年 12 月 8 日改定
2020 年 4 月 1 日改定

2. 学会誌編集委員会（会長指名の若干名で構成し､委員長は互選する）は学会誌の編集、論文審査の任にあたる。
3. 研究奨励賞選考委員会（副会長が委員長を務め、学術担当幹事と委員長指名の委員、計 10 名程度で構成する）は本会の研究奨励賞受賞者を選考する。
4. 若手シンポジウム委員会（会長指名の若干名で構成する）は学術担当幹事と共同でシンポジウムを企画立案し、運営をサポートする。委員の任期は 2 年とし 2 期限りとする。委員長は 2 期目の委員の中から互選する。

74

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2002 ń 12 Ɩ 1 Ƌżę
2003 ń 12 Ɩ 1 Ƌżę
2006 ń 6 Ɩ 30 Ƌżę
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Journal of Japanese Society for Extremophiles (2022) Vol.21(1)

編集委員会
委員長	
  ：石井 正治（東大）
副委員長：栗原 達夫（京大）
学会誌担当：相馬 亜希子（千葉大）、八波 利恵（東工大）、東端 啓貴（東洋大）
学会事務局	
 〒350-0815 埼玉県川越市大字鯨井２１００
東洋大学 理工学部 応用化学科 生命工学研究室 峯岸宏明研究室内
極限環境生物学会事務局	
  中村	
  顕	
  (幹事会幹事長)
E-mail: kyokugenjm10@extremophiles.jp
URL: http://www.extremophiles.jp/
学会誌編集
山本まみ
発刊日	
  2022 年 9 月 30 日