Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /0, No. /, ,2*,2/ (,**3) 67

280



 26 

 

) I²¬*, ** ³¯ ´* µ¶*+* ·, ¸*
¹ º* -'».* ¼½V/**** ^_`abc_** defghij *** ^kldMechanisms of Sulﬁde Components Expression and Structural Determination of
Substrate Precursor in Jumbo Leek (Allium ampeloprasum L.)
Ayumi Uchida*, **, Jun Ogihara*, Hitomi Kumagai*, Makoto Akao*,
Hiroshi Matsufuji*, Hidetoshi Sakurai* and Ichiro Sekiguchi**** Nihon University, College of Bioresource Sciences, +200 Kameino, Fujisawa, Kanagawa ,/,ῌ2/+*** Wadaman Science Co., Ltd., /.0 Karasumaoike Agaru, Nijoden-cho, Nakagyo-ku, Kyoto *.ῌ*2./*** Oukura Pharmaceutical Co., Ltd., ,1ῌ2, Midoriichi-chome, Sumida-ku, Tokyo +-*ῌ**,+ῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌῌ
The characteristics of sulﬁde precursor/components in the bulb of Allium ampeloprasum L. (jumbo leek)
were compared with those in other Allium species : A. cepa L. (onion), A. sativum L., and A. ampeloprasum L.
(broadleaf wild leek). Sulﬁde components from jumbo leek were analyzed by gas chromatography/mass
spectrometry (GC-MS) using spectral analysis. The contents of sulﬁdes from jumbo leek were dimethyl
disulﬁde (M-SS-M ; +.1 mg/kg wet wt), methyl propenyl disulﬁde (M-SS-Pe ; +/.1 mg/kg), propyl propenyl
disulﬁde (P-SS-Pe ; 1./ mg/kg), dimethyl trisulﬁde (M-SSS-M ; ,+./ mg/kg), methyl propyl trisulﬁde (M-SSSP ; +*.* mg/kg), and methyl propenyl trisulﬁde (M-SSS-Pe ; +..* mg/kg). The composition of alk(en)yl
cysteine sulfoxide (alk(en)yl CSO) from jumbo leek was S-methyl cysteine sulfoxide (MCSO ; ..+5*./ mg/kg
wet wt), S-propyl cysteine sulfoxide (PCSO ; *.+5*.- mg/kg) and S-propenyl cysteine sulfoxide (PeCSO ; ,..
5*.. mg/kg). The total content was the same as those of A. cepa L. and A. ampeloprasum L. Moreover, N(g-glutamyl)-S-(E-+-propenyl) cysteine (Glu-PEC, 21 mg/kg wet wt) which is a alk(en)yl cysteine precursor
from jumbo leek, was detected. Therefore, the lower odor producing mechanism in jumbo leek wasdetermined to be the same as that in leek. That is, the substrate precursor Glu-PEC is converted to S-E-+propenyl cysteine (PEC) by g-glutamyl transpeptidase. Then, PEC is oxidized to PeCSO, a substrate forallienase. PeCSO, another naturally present alk(en)yl CSO, is converted to S-alk(en)yl acid by allinase(C-Slyase), followed by sulﬁde production via formation of the dialkyl thiosulﬁnates.
(Received Oct. -, ,**2 ; Accepted Jan. -*, ,**3)
Keywords : jumbo leek, sulﬁde, S-alk(en)yl cysteine sulfoxide, N-(g-glutamyl)-S-(E-+-propenyl) cysteine῍ῌ῏ῌ῎ :   Sῌ    NῌgoῌSῌEῌ+ῌ <=>?0 @ABC DEFGHI10 
@G0qrYUZMG @Ustourpvw

L. Umnop8

JAKLMH =NOPQR=ST @EFxyH z={|}~ ,n9-,   ST \?R  !0=P"\ AXAruCp,/8

UVWX1YUZM ,**, A[M+ A\]^

CZCq0M  =#U$ AruH%¡     C-S

EF Xqe¢ :Jumbo Leek; Allium ampeloprasum£I18 zY£¤¥H =¦§¨Aru=©&ª«¬p8,/,ῌ2/+* !"#$%&' +200
0*.ῌ*2./ (%)(*+,-./01234 /.0+-*ῌ**,+(* +ῌ,1ῌ2 Corresponding author ayumi_u@mue.biglobe.ne.jp

*
**
***

­®  @ Allium sativum L. =GrAruH¯'=@)A°±X1 Sῌ





  ( : Sῌ  ῌSῌ¦ 

23ῌ4 : OP5&

 27 

,QRS1T6

281

alk en yl CSO U9 7 V 8 *W& 9 X : * ; YZ

'  GC & <1 K( LMN[\]R = 1+ W& ^

9_`

Shimadzu GC-+.A /a Flame Ionization Detector&b>?Rc')*;c<=1*de+fZ= FID g]61+ J & W DB-WAX ,/ m  *.,/ mm&h iiLDj@A&

Film *.,/ ῌmO#[UZ)*@

(kB&mCDn:)C <_op alk en yl CSO mC &=91b?*EFZ=- 

qb?&Gr*s+( W)&mC
x Glu-alk en yl C 78ῌ ( 9& ! :";*<=
h*y:KL*+MzNOt& ' Thomas

P{+|Q<=RS-U}~)*dJTUZ+C=0
V( @2A%#&WV8*,fZ=d$%x#\8& alk en yl CSO&=91b

 alk en yl CSO Dn:)CtuJvHB&IJtu=\ w! ῍S῍$ 'C l& GC-MS <1K*+0/

 .8*4&OP5u=iiL)+* &

|*< eB78 MzXYZ +* g *[+w # ῌLMM6ῌ]^( MeOHῌCHCl-ῌD.W. +, : / : - VῌVῌV /* ml, _ 32῍ ECSO 2`a)C + mmol

\#[3#%L GH 3#%L wNOPd ,*  %#1b< q @2A%#Ub?<=

cV dy

Glu-alk en yl C &/ &e<=D, L !"# / 

G+ 3]  w#. q fg*C+(h e 'o{.y*[.& CHCl-ῌD.W. , : + VῌV   jk &jkyD (i H#+

alk en yl CSO hy1

+&l*'+   

( 1 >#?&@2A%# $%& Allium ampeloprasum L. iiL mn& Allium sativum L. why*[ .& . mM 3-ﬂuorenylmethylcholoro
oD ,*῍,-  +* formate in acetone Fmoc

NOP pq& Allium cepa L. GH 3#%L $%&x#[ W&, ry*[+ -ῌ. &r

Allium ampeloprasum L. d%# $%& Allium

 stampeloprasum. L. &B78 @ MCSO S῍
S῍$%





)1 S῍ªM«ῌ ]^( ῌ uv acetonitrile ῌ D.

W.ῌacetic acid // : .,./ : ,./ VῌVῌV

'C @

(y¢_z£ &{¤A ' qalk en yl CSO u= S῍$¡[i[% d¦§y, w i  ECSOoD wxcr st &|y*[ .&¥w oD*¨<=1*dJ

Fmoc &D(E&) *C &FG

, HIJ+  ACSO &'( Yu}~y © w# *../ ῌm .&, .

x

y ¬y:LMN[\]# HPLC < B  PCSO &'( Ibert ) d S῍ªM¥ixB& alk en yl CSO & ­h HPLC &<

 PeCSO &'( Lancaster )3 &

*+



1K( LMN[\]1+R=( Shimadzu LC-0A

2, 

g]6( COSMOSIL῎ /C+2-AR g]x L£® 

B"ª 78 ,** g  *]^( +/* ml d2`ay diethyl disulfide + mmolD  , L ¯ ..0 mm+/* mm

'C g]60 ., /$ FLD ex ,0. nm, em -.* nm "ª : +*

&!"# , "ª U¥# [*_=qῌ l,  +.* ml ῌ min 0/ q°( Bu#ered±@O#[ p*²& , L  ] ³*°+´

acetic acid pH ...-῍ VῌV Acetic acid-+῍ VῌV

L $%x# U-c<= -1  + 8x

triethyl amine/MeOH/acetonitrile // : ., : ,./ VῌVῌV#[, Likens-Nickerson L-N ]^'

C+@LMMw CH,Cl, *dJ + 8  ]^

e

1

'C
.  !"#$% * CH,Cl, i (>v[%56(,µ¶#w#*dJ(0jk *
CH,Cl, , mlD/:1 GC-MS d GC *dJ·,- ./ῌ/ GC-MS &0/1K( LMN

alk en yl C&'() Glu*++ )C&@2A%#(4&OP51b ¸x¹%
&º)UCh&'C[\]R=1+ Yokokawa analytical systems HPῌ ©w #

/23* Series, g]6( J & W DB-WAX -* m*.,/ mm@2A%#& M z XZ 1./ g * 1*῍ MeOH +/*Film *.,/ ῌm

'C 0 .* /  0 .ῌmin :0 ,+* &1K0/ q0d

/0( ,** 1 2%$3 (4%56 Heml

BuOH h

D +* »7VR*¼½t&   .

*+ , - )(. ADVATEC
£®

NO, *+., J 1 NO

TUVU.W  /0 X  / Y ,**3  / Z

282

<

!"#$[2\] % &^<'( _`< ab , )c JC2de*f< &^ < MeOH + g +  ,9:&'(;2 + < GC =78

GC-MS >?7@ABCD

EAFG:A9:- &'( dimethyl disulfide M-SS-M : +.1 mg/kg wet wtn.o , ml 

FG:KipP:A9:&'( methyl propenyl disul-

HPLC / +**῍ MeOH 1 &': qr<

ﬁde M-SS-Pe : +/.1 mgῌkg wet wt Kis:KipP

2 LH-,* tuvw0 <:A9:&'( propyl propenyl disulﬁde P-SS-Pe : 1./

῎ tuv7 x

mgῌkg wet wt FG:Kis:y 9:&'( methyl

BuOH +2i

BuOH 2 d e h78

 28 

jklm



tuv Sephadex LH-,* &z:0k{|}Q

propyl trisulﬁde M-SSS-P : +*.* mgῌkg wet wt FG

 - cm!,0./ cm 2/] '~&^<w0 + ml 2

:KipP:y 9:&'( methyl propenyl trisulﬁde

MeOH  . ml ,.* mlῌmin  ) -/1

M-SSS-Pe : +..* mgῌkg wet wt =78AFG:y 92) UVmini +,.* 345[6 2/:&'( dimethyl trisulfide M-SSS-M : ,+./ mg ῌ kg< ]h &uQ0C ,2* nm = 2278:; . s Q x<῏HPLC 7 1

wet wt 9^<ABCD

9:&'(<=2>?w0 @    M-SS-P

AB2/ s Qῌ2tuv : COSMO-< < ;Ch]< E

SIL῎ /C+2-MS-II/ ῌm ..0 mm!,/* mm tu{}9Q M-SS-A A-SS-A M-SSS-A =78 A-SSS-A ]-. /D tuv2 : .*" 9E : ,/. nm

hPCPQ

HPLC

HI +*

 JCK: : ,** ῌl  +./ mlῌminHI : +*῍ NyPy:*.+῍ JK



GH R

@ ABCD

SQF<9:&'(<=  Gh]< <

L 9:&'(; @ =]hPCPQ<  Rt. MHI min /.1 N:s Q2 1

SQ G]^< O>nP7@QRL<, , ST"8GH R alk en yl CSO

1<STL C2nP < 0¡ 
UVWnP< aw0 X2 32./῍ 2T

ABCD

"

L : M N C 9 BCO : mῌz /*1** FAB-MS^2< < a<=PCPQ¢ GH R

S

h78<Q '~ "¥w02a¦bbQiyMu&T)*+  ,-. / UPLC-MSῌMS

R

UL alk en yl CSO PCPQ¢ GH

SQ@¤hG] ¢AB SX+*,A,Y£ 2/ Z[\ {|C

w0 + kg U alk en yl CSO ;2 + < JCK: : / ῌl :*. + ῍<2}yuFG:0uC TMS ;AFG:9:&+0( DMSO-d0ῌ 1 *.// ml 23456 NMRAB ECA-/**

Y£-

. / 2/]h78<
NMR  H-NMR dValue, /** MHz C-NMR dValue,+"#_

 ' >?E ABCDR PCPQ¢ GH

SQ G@ ¢+-+,/ MHz DEPT Distortionless Enhancement by Po-

<fh %~9:&'(;2dg  ABCD ©ª Allium ampeloprasum L.

 DNA   C-S  RNA =78 N Rh ,/ ¯°±¨ij²h]+ E a³k {´ C;

larization Transfer H H-COSY Correlation, Spectro-=78 lµH=  mnoG a§ ABCD

Quantum Coherence, HMBC : Heteronuclear Multiple

T%·bST q2w¸<

Bond Connectivity 278< = H-NMR  J - +)¹hº»<

ABCD

01!"#$%&&^<QRvw, F°: AFG:9:&+ 0(1 j }: Qii¼:v=78c

PCPQ GH R

 Glu-alk en yl Cp¶9:&'(¨῍ sqSTtu'()*+,-./+ "

C ®©^<scopy C H-COSY HMQC : Heteronuclear Multiple

J #Hz$%s Qr #ppm$!/**

]

?cS§ a9:&'(¨T<=EdE FAB-MS e8 UPLC-MSῌMS /]<w0/w«¬­1o NMR_  MCSO 2;   ` ¢MCSO PeCSO =78 PCSO 2;h]<

2/ tuv : Waters Acquity UPLC Shield RP +2
9E : ,/. nm, HI : +*῍ NyPy  ACSOh ABCDSQ =JCx< yzo{ABCDp¶|½

45ῌ6 : 789 :;<=>?@ABCDEF&

 29 

 +Tissues
Sulﬁdes NT



283


mgῌkg wet wt

Jumbo Leek

GarlicOnion

GH garlic

Leek

M-SS-M
P-SS-Pe
M-SS-A
A-SS-A
M-SS-P
M-SS-Pe
M-SSS-A
A-SSS-A
M-SSS-P
M-SSS-M
M-SSS-Pe

+41*
14/*



+/41*


+*4**
,+4/*
+.4**



+/140*+ -324-*
.3/4+*+ 0+04.*

,,43*





,42*
24,*


-42*
+-4.*
++4+*



,..4.*/-.4-*

3*43*+*14-*

,14,*

*43*
.43*



04.*


34.*
.40*
04.*

Total

1*4.*

- 03*4-*-34-*+ **.4+*-,40*

MeanSE n-)
Tissues
conpornent

Jumbo Leek

GarlicOnion

GH garlic

Leek

MCSO
ACSO
PeCSO
PCSO

.4+*4/,4.*4.
*4+*4-341*42+*+42+340.42+4*,43*4/41*42.4*+4*+14*+4*.04-.4,.4**41-41*4.
*4,*4,Total

040+4,++04-,+4.

+,40+40

0-4-/4,

143+4-

M, A, P, Pe 
 A : allyl, M : methyl, P : propyl, Pe : E-+-propenyl4

MeanSE n0) +  !"#$%&GHIC 21 mgῌkg wet wt JKL/NMR C +-C-NMR DEPT :M#N; O31PQR

ῌ FAB-MS NMR &3
FAB-MSST"U$*VWX/ Rt. /.1

 mῌz ,3+  !"#$%&'

 !"#[\]^_ Lawson *++T"U`aF&()$*+,- N-(g-glutamyl)-S-(E-+ -propenyl)cysteine CqrX/ sLi mῌz +./!"#./ LC-MS UPLC-MSῌMS 012 NMR &3+

H-

N-(g-glutamyl)-

S-(E-+ -propenyl) cysteine Y Z g-glutamyl) -S-trans- + propenyl-L-cysteine, Glu-PEC CbcX/ d +

efghiQR ST"UJjk^_/lmno p<tuv NM-S-(E-+-propenyl)cysteine Czc^_/,-

Glu-PEC `w  T"UJ,* mgῌkg wet wt xy

78J 2* mgῌkg wet wt ^*{ {#`._I[6

Glu-alk en yl C

T"UCxy78$*\]^_

  /0284 , 

 / ! ,**3  / " 30 



a : g-glutamyl transpeptidase, b : H,O,, c : allinase (C-S lyase), R : M (CH-), P (CH-CH,CH,), Pe (CH-CHCH-)#$%& '()*)*++ +%),-./012 3 4&+%) ,-./012 012

 6

DC Glu-PEC EFGHIJKLMJ( 

9:FGHEST> UVW&
Glu-PEC E@Z

BN ,-./012O5

8PQRC N-(g-glutamyl)-S-(E-+-propenyl)cysteine Glu-PEC XY(& go [\]^_`H

c.cdefgh UVW)iP .

57 89:;<=> ?@ AB C#e

g-glutamyl transpeptidaseab() S-(E-+ -propenyl)

cysteine S-trans- + -propenyl-L-cysteine PEC

glu-

jQkc.cdlmC n DlopC

tamic acidlW)*++ +%) ,-./012 Glu-PEC

FGHIJeq S-(E-+ -propenyl) cysteine sulfoxide>

PeCSO EthC W Sh(& PeCSO#SrEsC> AB
4& Lawson W u(&c.cdv fwx,

1e$%& Glu-PEC E ,,* nm

UV  E> #6 +  ()*
c.cdE .

u(&eQC !#opEZ)iP  Glu-PEC op>  .

    PEC 
> >D )*y$Fz2{H|0}~1
C-S lyase

8P Sῌ|cFz.i8,|2`FG1E)FGHEthC> "  ,

e +* Q(&c.c,-./012#$ *)  i*dwx,1e transo+o_^co|0`)%&k 7e9*SlFz.l +0 ' ()* &Z Glu-PEC # l)iP 4& ( )  *¡ transo+o _^c`Fz.KLMeq'()*++
()  transo+o_^c`Fz.l\ª«wx,1 in vivo e¬1­r ®2C>()*++ ,-./012 Glu-PEC 

3¯ *)()*#*l ¤¥<+*´5eµ6¶*. 

»8¢+,#9*£lq l)* ¤¥-¦eWF1§1KLMhkIJ
hE. C >¨/#©0 ,-./012 °4c±«²«³ *)FGHi8FGHIJh  E 7 ·(&¸

Lawson Wc.cd9:;*)'(,-./012 c.cdk GH ¹10ºd
P 012 ¼½C l89(&#ῌ :  rj|}'[B& 31 

-v -.rj|}B

a M-SS-M +.1 mgῌkg wet wt M-SS-Pe +/.1 mgῌkg wet
wt kg P-SS-Pe 1./ mgῌkg wet wt M-SSS-M ,+./mgῌkg wet wt M-SSS-P +*.* mg mg/kg wet wtM-SSS-Pe +..* mgῌkg wet wt alk en yl CS* Ba,-

MCS* ..+*./ mgῌkg wet wt PCSO *.+*.- mgῌkg
wet wt  PeCSO ,..*.. mgῌkg wet wt zy ( B)a`rj|}~mno*. Glu-alk en yl./C  Bx Glu-PEC 21 mgῌkg wet wt+, -.zya 

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2

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Ariga, T., Kumagai, H., Yoshikawa, M., Kawakami, H.,
Seki, T., Sakurai, H., Hasegawa, I., Etoh, T., Sumiyoshi,
H., Tsuneyoshi, T., Sumi S. and Iwai, K., Garlic-like but

285

Odorless Plant Allium ampeloprasum ‘Mushuu-ninniku’,J. Japan. Soc. Hort. Sci., 1+, -0,῍-03 (,**,).
 &' ()*+,-.
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LINNE Shiro ;<=>? @ABA .0 -0/῍-1*
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YZ[+\-]^+_`a 2b1 // /.3῍//2
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cdef gh i !" / /$#jk l mno S῍pq/"rst $%o HPLC &u 2b1 /, +0*῍+00 ,**/
Yu, T.H., Wu, C.M. and Liou, Y.C., Volatile compounds
from garlic, J. Agric. Food Chem., -1, 1,/῍1-* (+323).
Ibert, B., Winkler, G., Muller, B. and Knobloch, K., Quantitative determination of allicin and alliin from garlicby HPLC, Plata Med., /0, -,*῍-,0 (+33*).
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the S-alk(en)yl-L-cysteine sulphoxides in onion (Allium
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Thomas, D. J. and Parkin, K.L., Quantiﬁcation of alk (en)yl-L-cystein sulfoxides and related amino acid in alliums
by HPLC, J. Agric. Food Chem., .,, +0-,῍+0-2 (+33.).
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Natur. Produ., /., .-0῍... (+33+).

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