Abstracts
rBJ]!. Res. Inst. Ferm. Yamanashi Univ. No. 11, !'iov. 19(14)Studies on Malo-lactic Fermentation
Isolation of Malo-lactic Organisms and Inocula-~ion of theIII.

Isolates into Musts
Hideo NONOMURA, Yuwao OHARA, Hisashi KAGAMI and Kei-ichi KAZAMA:
NiJ.>p:m Jozo Kyok.1i Zassh£ (.1. Soc. Brew. Japan) 59, 513 (1964)
Three hundred and twenty four strains

10 were inoculated into th~ red musts (Black-of malo-lactic organisms were isolated fromQucen and Merlot) and the white musts(KOS~1U) respectively,26 sam;>les of musts and lees (representing7

each must was 20 lin \"olum~, and contained 50 ppm S02, 23varieties) with ROGO~AS tomato

grape

juice-liver extract medium modified by theor 25 % sugars and 2 % starter (ca.

.additioOl of 20 ppm

celh/lIll).

20 ppmactidione,J O·A strain of heterofermentat-ive

eurocidine, 500 ppm sorbic acid and 2 90

Lactobacillus and a strain of Leuconostoc.agar.

(sp. related to citrovonllll) were effectively

The ratio of the organisms to total

isolates increased in the progress of must

induce the fermentation in all t!1e threefermentation.

And it was usually greater

kinds of musts, and the latter strain seemedin red wine lees than in white examined.

to cause the fermentation with the greatest

repres~ntatives,

The

isolates

were45 strains,classified

intoof4regularity.

thegroupsstrains, were lower in acidity, higher in(included 9 isolates) of Lactobacillus, and6(36isolates)ofLeuconostoc.

The wines from the musts, inwhich the fermentation took place by these

pH \'alue, and better in flavor and tasteNine

than the control wines.

strains representing 8 groups out of above[Bul!. Res. Inst. Ferm. Yamanashi Uni\'. No. 11, Nov. 1964)

Studies on Japanese Wine Yeasts
VIII.

Cla,sifica'::ion of Flor Yeasts Isolated in Yamanashi Prefecture, Japan.
Isami YOKOTSUKA and Shuki MOROZUMINippon Jozo Kyokai

Zasshi (J. Soc. Brew. Japan) 57, 836 (1962)
Thirtyyeasts

werethreeThey were classified in accordance witht;,pical strainS of f10r

selectedfrom847

the procedLHes and systems ofcultures

LODDER

andisolated from 470 samples collected at C6K. -van Rr,J as follows: 5 strainS of Sacch-wineries and several vineyards

arolllyces cel'evisiae ; 6 of S. l'osei ; 2 ofin grape

S. steineri ; 2 of S. exiguus; 3 of S.

and wine producing areas.
77

Abstracts

78
helerogeniclis and 15 of S. ovijormis.

Protuberances

resembling

toconjugation

Following two noticeable characteris ics

tubes were always present on the cultures.

(1)

on GORODKOWA agar slallt or gyps:Jm block.

were observed for all strai'lS tested:

fseudoffijceliums were not form~d and (2)IX.

Physiological Properties in Fermentation Stage ofFlor Yeasts Isolated in K6fll Valley, Japan.
Isami YOKOTSUKA

The com;>aris::>n of physi::>l. properties

Ibid., 57, 943 (962)growth or

ferm~nation,thei r tolerance toin fermentation stage between f10r yeasts

S02(I) and non-film fvrming yeasts (II) was

velocity, and their abilities of producing

conducted by using 33 strains of 1 and 51

alcohol, organic acids, esters and aldehydes.

orpH

values,theirfermentationof ordinary II isolated from many wines

But the results of testing of their wines

m'lde by spontanous fermentation.

showed that bad yeasts seemed to be more

The data obtained confirmed the resJltsof many workers,and indicated that 1

abundant in 1 than in II and tJ1at good ones.
were more in II than in I.

resembled ordinary II in their temp. forX.

Utilization of Flor Yeasts to Flor Sherry Making andImprovement of the Quality of Dry Table Wine.
0) Flor Film and Its Cells of Flor Yeasts.

Isami YOKOTSUKA:Ibid., 57, 1197 (l962)

Twenty nine strains isolated in Kofu

(2) Excessive film formation occured f[(qu-valley, three (Jerez-5, Xeres-l and Xeres-2)

ently on wine contg. below 14 % EtOH

of typical Spanish and two (D-Douroveu

under favorable environmentaland E-Evora) of Portuguese f10r yeasts werewhich deteriorated the quality of wine.

cultured on dry wine, and film formation,(3) Cells of film. did not essentially differ

type of fi 1m and its cells were observed asfrom that in the fermentation stage.

condit!ion,

Theyfollows; (I) Each strain had a special type

gathered into a flock, but did not form aof film which was classified into five groups.

pseudomycelium.

XI.

(2) Changes Brought about in the Composition
and the Flavor of Wine by the Flor.
Isami YOKOTSUKA:

Twenty nll1e strains of Japanese andIbid., 58, 82 (1963)
compn. and the quality of wines by the flor

fi ve of foreign yeasts mentioned above werewere estd. as follows: There was no much

cultured on dry dessert wine and dry table

noticeable differece in the changes brought

wine, and the changes brought about in theabout in the compn. of wines by tbe f10r of

79

Abstracts
between the Spanish yeasts and the othel s.

between 0.5 and 2 gjl.

Flor character developed noticeably and thethe cultures did not change so much.

al dehy de (1) increased to between 150 andamount of sugar used by the flor seemed

250 mgjl by the fourth to fifth week. The

to be very little.

strength of flor character increased as theacid or lactic aci d wereamount of total 1 increased.

carbon source for film growth. Total esterS

But it is

The pH values ofThe

EtOH, glycerol, acetic
likely a goodlikely not the amount of total 1 but theincreased a little, but the amount of neutral

amonnt of fixed 1 that has strong effect.

esterS did not almost

The amount of EtOH used by the f10r wasincreased remarkably and in some case it

only below 1 %.

amounted to about 800 mgjl after one year.

In most of the cultures volatile acids
decreased to below 0.2 gil one year after
inoculation.

Fixed

1 increased

Total Ichange.

inwineastotal I

increased, but not in a constant ratio.

A fall in fixed acidity ranged

XII.

(3) The Influence of Environmental Factors and the Composi tionof the Wine on the GrJwth of Flor Film: Culture Temperature,and Alcohol Content and pH Value of the Wine.
Isami YOKOTSUKA and Shoji GOTO

The influen"e of incubation temp. , theale. content and the pH value of the win~50

andIbid., 58, 169 (963)

r e

all

complet~ films,strains

didnotformand 1, if it o~curred, wason fi 1m formation (1) of flor yeasts wererestricted to scant_, small patches. (2) The

examd. und~r lab. conditio.~s using 27strains

ale. strength of the wine : An optimumofJapanes~

montio:1edand fiveabo/e.

of foreign

Theresults

yeastso)tlin~dwere as folloNs: (l)Temp.:On table winosale. co:Hent for 1 by most strains used wasb~twcen 13 and14%, while the max. for

com::>lete films with some strains was above

conig. less than 13 % EtOH, the rapi d 115%. In this re;pect four strains resemble

o:cu ned at temp. up to 28°C with mostquite closely

strains uSed. It has been proved by mmy

sherry yeasts. The rate of 1 was increased

inves:igators that the o::>timum temp. forby low ~ring the ale. content, but 1 at such1 on wines contg. between 14 and 15%

low stren~ths were t·')o m:.Jch, very unstable

EtOH is about 20°C,MARClLLA'St;picalSpanish

and that at temp.

and apt to sink. (3) The pH value of thehigher than about 23° to 25°C, 1 does notwine: The optimum pH values for 1 wasoften oc:ur. At ION temp. beloN about 150

between "3.0 and 4.0. As the pH values ofe, 1 was usuall_ very solw. At about 100emost Japanese ordinary wines are between

on wines contg. les; than 12% EtOH, one

3.0 and 4.0, so it is unnece;sary to adjust

thi rd of the strains usetl fai led to develop

the pH value for 1.

as a fi 1m and at temp. do .vn to between

XIII. (4) The Quantities of Sulfur Dioxide and Sugar in Wine.
Isami YOKOTSUKA, Shoji GOTO and Yoshihide YAMAKAWA:Ibid., 58. 1221 (1963)

Abs~racts

80

The effects of the quantity of 50 2

below O. 2 %.

usually(4) Good 1 wasadded and sugar in wine on the film for-observed on the wine, which contd. onlymHion

below 0.1 % of reducing sugar estd.

(I)by flor yeasts in Delaware

white table wine contg. below 12% EtOH

BERTRAND'S

were examd. and the results obtained werelacking in fermentable sugar.

methodaDdbytherefore

was0 active 1

as follows : (1) The lOxity of 50 2 on 1

was caused byadJing sugar in the wine on

was ve7 strong. It could not be che<ked

whIch flor yeasts had completely developed,by culture temp. or ale. concn.

of wine.

but it occurreJ when~mallquantityof(2) The tolerance of 1 could not be raised

new wine contg. almost no sugar.

by training.

dingly there seems to be better growth(3) The quantity of sugar

used in 1 on table wine was so sm dl, andAccor-facto rs than s;.tgar.

XIV. (5) The Concentratio:J of Ethanol in Wine.
Isami YOKOTSUKA, Sl-oji GOTa ancl Yoshihide YAMAKAWA
I!Jid., 59, 84 (1964)
The effect of the strength of EtOH ona II wines tested were w much that they

1he development of flor character and theamounted from 60 to 70% of their initial-changes in the com;lOnent of wine broughtamounts.

<lbout by flor W;.IS studieJ. The flavor wasvigorous flor seemed to be check the flor

tastedto consume aci ds in wine.

and components were analysed 0:1

The

strength of EtOI-I

ur.der(3) Both thefour kinds of dry white wine; contg. 15.0,

accumulation velo~ity and the accumulatedb~ing

amount of aldeh, de (I) under vigorous flor

stored under the film; of ]erez-5, WF-I07,

were generally not a(fected by the strengthVV-21O and W-500 for one year respectively.

of EtOI-I. (4) The flor character (flavor)

14.3, 13.5 and 12.5% EtOH, after

Results obtained w~re as follows: (1) Theof the wines under flor increased to someamount of EtOH consumed by the flor wasextent as the am'Junt of 1 increased. It was

SJ small that it was not a seriDus problem

found that the amount of 1 accumu lated didfrom econo.nical point of view.
tota I(2) Bothaci d s an d non-volati Ie aci ds wereconsumed the mo,t by the f10r in the wines

notdominate the flavor,flavor

dependeJ

ontheand tlutthestrains.

Theamounts of I, in which superior fla\'or badcontg. 15.0% EtOH and the least in thealwdYs devdoped under the film of ]erez-wines contg. 12.5% EtOH. The amounts

5, were a Iways less than those in the winesof volatile acid; cor.sumed by the /lor forunder the fi 1m; of the others.

Xv.

(6) Culture Temperature.

Isami YOKOTSUKA, Shoji COTa ancl Yoshihide YAMAKAWA
Ibid., 59, 163 (1964)
The effect of cu Iture temp. on the qua-flavor

and analysing the components oflity of wines: under flor was investigatedwhite dry table wines contg. 12.8% EtOH,

by observing the fi 1m formation, tastingon which the films of ]ere--5 and WF-107

Abstracts
had covered for eight months at 10°, 15°,

the accumulated amount of aldehyde increa-20° and 25°e. Results obtained were as

sed as the temp. rose. (3) After one month(I) No film formlt'on at 10 0 e.

follows:

Slow and a little formation at 15°e,
~I81

but

fJor chlfacter was developed in all wines
except th~ wine incubated at 10 0 e. Thecom;:Jlete film cover was never made throughflavors of the wines incubated at 20° andthe whole period studied.

25°e were equally strong, but that of the

Very rapid at

both 2()0 and 25°e; com;:Jlete film coverwine at 15°e was comparatively weak.

after 20 days and thick films after 30 days,

After 40 days they were all satisfactorilly(2)strong, but after 50 days the flavor of theThere was no effect of temp. on th~ con-wines incubated at 25°e, smelled of yeast

sumption of EtOH by the flor.

and was inferior to the other wines.

and then the fi Ims grew to:> much.

sumptiontemp. rose,

The con-It isas

worth to notice that sufficiently strong £lorbut no remukable difference

chlracter was developed in the wines onot volatile

acids increased

was recognizeod in the amount of total acids

which the film were only 10 to IS};:).

consumed. Th~ accumulation ,'elosity andcomplete thin film cover seemed to be best.

TheXVI. (7) Strains of Flor Yeasts.
1sami YOKOTSUKA, Shoji GOTO and Yoshihide YAMAKAWA

Ibid., 59, 267 (1964)
The effects of 27 strains of Japaneseyeast~

we3.k, could not de lelo? strong fragrantmentionedflor ch3.racter, and mOit strains of Japanese

before, on the quality of both dessert wine

ye3.sts tested fell into this category in theirand table wine under their films for 90

abilitie3. (2) Only the strains, which oxi-and threeofSpanish

days were investigated by tasting the wines

dized alc. to aldehyde, and aldehyde toand analysing their components, and th~acids to ade:juate degree3, and then de-conclusions were obtained as follows: (1)compd. acid, moderately, much improvedTh~ straiQs of th~

yeasts,whose oxidative

activities and de~ompg. abi lities

the qualiyt of the wine3 by reducing theacidsacidity

at their film stage were too stro:1g or toofla\ or.

0:1

anddevelopingstrong

fragrant[Bull. Res. Inst. Ferm. Yamanashi Univ. ~0.11, Nov. 1964)

Studies on Japanese Plum Liqueur
1. The lnfuenct' of Manufacturing Processes upon t1:e Quality andthe Composion of Organic Acids d Liqueur.
1sami YOKOTSUKA and Shoji GOTO : Nippn Jozo Kyckai

Zasshi (J. Soc. Brew. Japan) 59, 633 (1964)
Six kinds of the liqueurs(I)were madeby the following different processes:(a)Moderately ripe plums and sucrose wereput in approx.

:~5% EtOH for about twoAbstracts

82months,then the extract was se;;arated,bottled and matured at room temp.

(b)always detected in all I. The conte:lt ofmalic acid was considerably affect'ed by theAfter plums were put in 35 % EtOH forpro:esses. (3) The kinds and the amounts

two months, the ext. was drawn off, thenof II detected in I by column chromatog-sucrose put on the same plums fot' the sameraphy were approx. as follows

perioJ, and the ext. se;;d. Both ext. werein % to total acids) : citric acid 55, m"lic

blended, and the mixture w dS bottled andacid 33 as a melD value, uniden.tified acid;matured.

(c) Completely crushed plums

were used.

(d)

EXln.

approx. 50% EtOI'1.

(e) Over-ripe plums(0 Extn. W.1S done for one

were med.
ye~r.

was done with(expre;~ed

10, oxalic acid 2, and acetic acid 1. 5.
(4) The quality of I examd. by tasting WdS
as follows : In 10 months after bottl:nJI (a) was ordinary. I (b) was the best. IThe compn. of I produced was exa-(c) was somewhat too sour and rough in'mined by a general chern. analysis, organic

taste and smelled of seeds. In 20 months

acids

after bottling: I (c) had improl'ed consi-(II)in I were detd. by column

chromatography, and I tasted. Thus thederably in quality.

Ide\'eloped stron:;

1nflue:lce of the proce;' e, appLed upon thefragrant flavour,_compn. esp. of II, and the quality of I

and rounded in taste. I ( b) was a lmost as

we e stud ied. The resu Its obtained were as

good as I

follows:(1) There W"s only little diffe-bnd was rich in bod;(c), but it was somewh'lt flatand watery in taste. I (e) developed strong

rence in the yields of I. (2) Considerable

fragrant £la\ our, but was fairly cloud; inchanges were oberved in the amounts ofappearance.

Both I (d) and I (f) hadII; exce,)t malic acid, prese<1t in I. Theiralways fairly cloudy appearance and badamounts depe'1ded to some extent on theoff-odors.

proce;~es,but the same kir.d; of II wereII. Identification of Two Unlmcwn Organic Acids found In Japanese
Plum Liqueur.

Isami YOKOTSUKA and Shoji GOT0
Two unknown organic aci d; (I) fOend
In the lique'Jr were studied by paper cl1ro-Ibid., 59, 636 (1964)

and its isomer, respe:tively.
Next, II was synthesized and its purematog raphy, column chromatography, gas

crystals were o~tained and demonstrated.

chromatography,Then the same expts. as mentioned above

analysis.

andinfrared

spectrum

The identification of the neuteal

compounds fro,n I,

and the comparison

weH',( repeatedusingpurecrystals

ofsynthetic II, and the data obtained in thebetween the amount of alkali used to neut-former e'xpts. and in the latter ones wereralize definite amounts of I and th~t medcompared. Thus it was con:luded th'.lt I

to saponify themwere II and its isomer,

were also made from

res;Jecti \ely. II

resu Its obtained in these expts., these two

was found for the first time in the liqueur

I were presumed to be monoethylcitrate(II)in this studv.