食用キノコのスフィンゴ脂質 : Ⅲ. オオヒラタケ Pleurotus cystidiosus のマイコグリコリピドにおけるヒラタケ属共通の多分岐型糖鎖構造

全文

(1)Shiga University. ]^XS_`SN8u vwoS No. , pp. ῍ , .

(2) ῌῌ ῎ Pleurotus cystidiosus . !"#$%&'( * ῌῌ ) *. +** , - . / 0 , 1 2 3 4 , 5 6 789 : ; < = , > ? @ A , B 6 C D. Studies on Sphingolipids of Edible Fungi ῌῌ ῎ Mycoglycolipids of Pleurotus cystidiosus EOhiratakeF have a Common Polytomy Sugar Chain among Pleurotus sp.* ῌῌ Saki ITONORI **, Kouji YANO, Hisao KOJIMA, Yukiko KURODA Masahiro ITO, Kenji YAMAMOTO and Mutsumi SUGITA Abstract Edible fungi, mushrooms, are a popular food in Japan and over fifteen cultured mushroom species are available at the food markets. Recently, constituents or ingredients of edible mushrooms have drawn attention because possibilities have been seen for their medical usage. Mycoglycolipids EBasidiolipidsF of higher mushrooms have been characterized as glycosylinositolphosphoceramides, and they have a common core structure of Mana ῍ Ins ῍GPOH῍Cer and extension of Man, Gal and/or Fuc sugar moieties. Five mycoglycolipids were purified from an edible mushroom, Pleurotus cystidiosus EOhiratakeF by successive column chromatography on ion exchange Sephadex EDEAE῍SephadexF and silicic acid EIatrobeadsF. Their structures were characterized to be Mana ῍ Ins ῍GPOH῍Cer, EGala῍ FEFuca῍FGalb῍Mana῍Ins῍GPOH῍Cer, EGala῍FEFuca῍FGalb ῍ Mana ῍ Ins ῍ GPOH῍Cer, EGala῍FEGala῍FEFuca῍FGalb῍Mana῍Ins῍GPOH῍Cer and IGala ῍E or F Gala῍JEGala῍FEFuca῍FGalb῍Mana῍Ins῍GPOH῍Cer by sugar compositional analysis, methylation analysis, periodate oxidation, partial acid hydrolysis, enzymatic hydrolysis, immunochemical analysis, gas῍liquid chromatography EGCF, gas chromatograph῍mass spectrometry EGC῍MSF, matrix assisted laser desorption ionization time῍of῍flight mass spectrometry EMALDI῍TOF MSF and H῍nuclear magnetic resonance spectroscopy ENMRF. Ceramide constutuents of their mycoglycolipids were composed of phytosphingosine as the sole sphingoid, and mainly ῍hydroxy C : and C : acids as the fatty acids. Key words : Mycoglycolipid, Glycoinositolphosphosphingolipid, Edible fungi EPleurotus cystidiosusF. * ? K L M N O P Q R ? % S T U T E U VW XYF Z[\ ]^XS_`SNaS_b EDepartment of Chemistry, Faculty of Liberal Arts and Education, Shiga University, ῍ ῍ Hiratsu, Otsu, Shiga ῍ F cdeXSfghiSNfgjkS_b EDepartment of Bioscience and Bioinformatics, College of Information Science and Engineering, Ritsumeikan Uni- ῌ. ῍. versity, ῍ ῍ Nojihigashi, Kusatsu, Shiga ῍ F lmXSXSnjdoSKLo EDivision of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Oiwake῍cho, Kitashirakawa, Sakyo῍ku, Kyoto ῍ F ** pqr῎)* + ECorresponding author : Saki Itonori, E῍mail : itonorissue.shiga῍u. ac.jpF E U V RthF. NII-Electronic Library Service.

(3) Shiga University. ±b. ]²cd ³´²ef µg²¶hi·¸²¹j kº²»; ¼K²lh m. }> ~6$?97 àp3( ῍ . . y5>3 T

(4) ;6 &'HCI>>JK(`v+o.

(5) . u,. .

(6) . !"#$. ῎ . . %&'()*++, -. /

(7) 0'12

(8) . ῎. ῍. . 34(

(9) 5!67#$+3(3. RRT

(10) ; Pleurotus cystidiosus . 2+, 4(1 89. ? L@AZBGM g (CD. #$:

(11) ;(<=>; Agaricus bis-. LD? gM EFGHI. porus :

(12) ; Agaricus campestris (?. f FPJ#$.,. @AB!CDEF > @GHC I>>JK LMycoglycolipidM #$. ῎. ῎. > AB()N1O 4$(. ῎. ῎. ῍

(13) .

(14) . -$ GalaῌGal O+ GalbῌMan. CD L gM F1[.3. (PQR>S!TEUVW#$. (K7 4$$ L (=gg /. X! Y#$+ L ῍ ῌ ῏ M,. F / L L : : , v/vM 1 , h. AB!(Z(["#$%. &Pi , h (M7., !(M7. +\+\&9'(]$&+( )^_. NOP QAB7. àB1Ob*(cd1O, e. R g S., +1 S$.QAB. (AB ?I>fghAB&. 7¡z >L7¢ T+¡@£-¤. Pijkl ShABCDEF >. zD h&Piz ;¥ hI>fgA. @G@jkl SAB!*$+. B¦.m zfE UV Ljkl SA. , + mn(CDEF > @G@. B7M g S. LD?W.. AB,o.PQ+(HCI. §M,. >>JK 4$(-93./. ῎. ῎. ῎

(15) . 0$12 p3à qr!4s$. jkl SAB7 g DEAEῌSepha-. +, (\ àp3 t-. dex Aῌ ¨CR ©ª

(16) =gHEI

(17). 1 Y#$+u(HCI>>J. klF LCOO« h g x cmM . Kv+ Mana ῌ Ins ῌwPOxῌCer !. P N7NO

(18) XY( ZX. y5zp3%!{#$. (=gg / F / L L : : ,. + L ῐ ῌ ῍ῌM, T

(19) ;|T

(20) ;67 #$. v/vM ZX(F &Pi ZX(. M [@z ¬¥ LF NM. T

(21) ; Pleurotus ostreatus &Pi. ®

(22) + &Pi\CR hAB. }> ~ Pleurotus eryngii !*$+!. 7@AB77. L῍῎M, (. 4$(HCI>>JKv+ T

(23). =gHEI

(24) klF1 ]Kn(=gg . ;KC<&Piz>(qrnP. / F / L&PiF PN7. -. }> ~8nPqr!$. P ^¯(_°`B . 4$(àp3!?97&+):. mg LD?W. §M (&Pi\. ! Y#$+ L ῏ ῍῍M,. CR hAB7 )a mn( . ;<=1 3Q) (T

(25) ;6(. M [@z ¬¥NOPN7P. 1ORRT

(26) ; Pleurotus cystidiosus. ^¯(°`B g LD?W. (HCI>>JK1 T

(27) ;. . §M (@AB7 LHCI>. NII-Electronic Library Service.

(28) Shiga University. º wB#t»¼'(.

(29). jsh. . !D> ?. GC Shimadzu. GCῌ A {P {. ῍. ῎. - .

(30) . mm x m #"& jsh. . BF mm # hw. ῌ

(31) / / :. Shimadzu HiCapῌCBP c . : , v/v ῌ RSῌ. {- o o o/min !. . . x cm !"#!. $. mg #%&'(). ῍. ῐ. ῍ + , +-. &'()*. *+#,-. !/012#34. mg #i! mL # M l%)m.. , 5. ,6- mL 789 . . h, o ) $K. qr '. frs. : ! ;-. . % mL # nῌvwx,yz . |. <=> ?@!ABCB#DEFG. 6 o. +. H ?50IJ

(32) #'(>#. U |-. +!%F. TLC KL M#N #O!/P?Q. %P hk&! h 5?!. fr. R/S fr. -TUVU AGLῌ .. % 5. mL # F¡. AGLῌ >W fr. > fr. *+-. X. . . #. 6 mM %F. # ῌ

(33) / . 6 )m? h, o !' $. / : : , v/v ,!/P. 6!(¢%)m. £¤#F¡. ?. TUVU AGLῌ a R/S AGLῌ b Y. . Z fr. *+-X . o + U | ! . $ K. jk)m[G+. # ῌ

(34) / / :. mL #¥R/S mL #"¢%). : , v/v ,!/P?. AGLῌ YZ. m?. o, min ¦s !D)Z. [R. 57U#X . ? GC {P GC {-. . x cm \. ] ml/min >^0. . o o o/min y5?^§. _!)`, 5. ?jsh !D#¨>©ª« P. ῍. ῏.

(35) TLC. ῍. ῐ. ῎ /. TLC

(36) a-. E. Merck bZ# Silica gel. 0+. , . +-.. &'()*. 5 -. ῌ

(37) / /. mg #i9 . mL #¥js. : : , v/v c . . t¡wh)m K. $i¬n&. def Q,-. gh / %. ?*+*®+¯°!/P?!$. i . DittmerῌLester i ῌ῎. ± ²U!, mg #-.%F. R / S HanesῌIsherwood i ῌ῏. > mL #³Fjs)m. jsF P ῌῐ min K. $6p ?. !/P. *+. mL #´ ¡ > mL #) ῍. ῐ. m?/µ!0 . ¶·1!/P?Mµy.

(38) GC. ῍. ῐ. ῌ . !"#$% !. #!. &'()*. z µ#´ ¡ µ ?. jsF2 ²#j. mg #i! mL # M jk&. sF2! mL #¢% / l% / : :. l%)m?n . o # h ). , v/v )m. 3¸+ 3¹a¥. TO-. pK. qr '%jsItu. SHIBA ERῌVS sec e¯° ?$. mL # nῌvwx,yz KL. K ῌῑ. . #'%{c |6!}%~)m?. |-. +!hk&!5?. % K. . +U |. !% K. mL # F¡. NII-Electronic Library Service.

(39) Shiga University. ®. /¯#$ °±¯²% ³&¯´'(µ¶¯·) *¸¯¹+ º¯»' ,¼. mM . ῍. ῑ. !"#$#%&'()*.

(40) , h

(41) . + , - . /

(42) MALDIῌTOF.

(43) !. MS. '(). AGLῌ AGLῌ b AGLῌ SOT AGLῌ.

(44) *+,-. /0 1 GC 2 3. "#$%&. MALDIῌTOF MS 27z Shimadzu/. 45 6 /min 78. KRATOS KOMPACT MALDῌ

(45) {F;|. . SOT}~BVF-7z Sun Micro-. ῍. ῏. ῎

(46). systems Workstation SPARC station. /9 . . :;<=> nῌ?@.

(47) >U-27z KOMPACT UNIX. ABC0

(48) GC 2 345 6. software

(49) R D

(50) mg/mL . /min 78 . 7O7UVV- / :;<F- /. ῍. ῏. ῏

(51). : : , v/v 7 ABt-t. mg D 7 mL E:;<F-G. vF7bc D mL

(52) % Z.

(53) h, ῌῑ H. '*t4o. I. > ῌaminoῌ ῌmethylcoumarin, cou-. J

(54) :,-K>L-

(55) nῌ?@AB. Q7 mL euU. C0MN ! )OP:;<F-

(56) N. marin SIGMA

(57) mg/mL K. QR mL M . ;<F-1 bc w. / :;<F- : , v/v SOT .

(58) 4oFuU>. mL UVV-

(59) ! WX. % Z! MALDIῌTOF MS 2

(60) . Y7 $YUVV-. vFFz nm "vFF

(61). Y

(62) Z'7 mL / :;<F- : ,. R zoyBF i. v/v 7O[\ ! "#$]. 7zK4 Pseudopotamilla occelata E. %&. '(. Ji AGLῌ mannosylinositolphospho-. a

(63) :,-=-.1 GC 2. rylceramide MῌHh m/z SOT. 6 /min 78 . AGLῌ dimannosylinositolphosphorylcer-. ^_Gà

(64) bc. amide MῌHh m/z

(65) R ῌῒ . ῍. ῐ.

(66) GCῌ. Q. 79 . MS. xHI

(67) ¡¢£¤7z. Shimadzu GCMSῌQP d>UVef. c. !7D

(68) b. ! ¥9euU>

(69) mL . 4ghi2j7OP Qkl$2. 2¦§z Applied Biosystems/Voy-. 234῍Shimadzu HiCapῌCBP ;. ager System

(70) {F;27z Work-. 345 ῍ :,- *-mF-*+. station Ver.

(71) R. L F 2 min 6 /. FR i7z@B¨K Lucilia. min 6 /min ; J

(72) 2 . caesar EJi ceramide triῌ pentaῌ. zoyB. min 6 /min 6 /. heptasaccharides m/z . min ; ^_G`2 min 6 .

(73) R ῌΐ . /min 6 /min 7n ; oB. ;FgpF>5῍ ; D qr5῍ ; ?s῍ kPa ; >tuv >wx῍ min ; oyB s῍ eV EI ; oyB ῍ mA EI . ῍. ῒ. 0123*4567 ManῌIns 8 9:;

(74). mg D AGLῌ

(75) mL K ; <F-7. ©(7 mL mM . M ª« pH

(76) h " ¬Z mL K,vBfF-

(77) . NII-Electronic Library Service.

(78) Shiga University. PI2Ng»

(79) ¼R.

(80) . L h k l d῍῍ ῎῍e . mL . . !"#$ %&'(%)*. mL 2qrr'K, / GHIJK d : , v/. mL 2 4 a66. +(,- h ./01 . ve GHIJK GHIJK / d : , v/. %23456!7!"89:;<. ve 2st SepῌPak fJ*. = >?01 ?. +n. @ 56ABC MALDIῌ. Waters je. dSepῌPak Plus tC Cartridges,. fJ*+n . TOF MS ? #1D E2F2. mL 2st mL 2GHIJK. mL 2 M GHIJK: L. RTkl T mL |. h GHI+MN nῌOPQ

(81) R

(82) :. KMIJK / :y #" TLC spot-. STUVWX%Y<@Z[. ting test 2 @. . 56 \ABC ]^"+. ῎. ῍ῌ. ῎.

(83) :UV_" ` a2@W. bῌ

(84) . !" . #$%&' . \. mg 2 y mg 2 H ( r P M. U 2 Kf+'NghHJi dEscheri-. JK:)*+(, mL 2bc. chia coli SigmaῌAldrich je . d M q

(85) :+

(86) :)*+(, pH e. M 9:

(87) (,bc dpH e. L h kl X%Y<. \ E6. U d mLe 2%. @Z[kl mL 2m+n

(88) . L h k l d῍῍ ῎῍. mL 29: L min. ῎῎e 2. . . opK& opK&. Pak x01. mL 2q. ¡¢ Sepῌ. rr'K, mL 2st <u 2qrr'K,u GC v#w GCῌMS ? x d῍῍ ῍῎ ῍ῐ ῎ῌe. ῎. ῍῍ ("$) £Npqyz{= mg 2y mL 2nGpKNK'PM¤ }~x. ῎. ῑ. ῍. \ mL 2g&%: de. HῌNMR . mg 2yyz{. | mL. L h kl . 2 DO/DMSO }~ #1. ? ? @ \. \kl ? JEOL JNMῌALPHA. ABC d῍῍ ῎῏e. ? MHz. y L AGLῌ d ae. ῎. ῍῎ * B +,-*. /0 1 23. AGLῌ d be AGLῌ d e AGLῌ . *. 4567 TLCῌimmunostaining. AGLῌ b AGLῌ

(89) . y£Npq¥£¦J* dPolygram. . Sil G ; MachereyῌNagel GmbH&Co., Germanye. ῎. ῍ῌ. - dy2- Ty. aῌ bῌ

(90) . 2KMIJK / :y§῍ mg Immu-. . nostaining § ῍ mge ῌ£ r ¨. ῎. ῍ῌ. ῍. aῌ

(91) . SigmaῌAldrich . IJK / /

(92) d : : , v/ve © ª>«¬ «¬. !V£¦J*. Z. PBS d mM +. mg 2 y mg 2 H ( r P M.

(93) :bc / 3 pH e N£¦J. JK:)*+(, mg 2 gῌq*I. ®¯ °! " ±²³rP. q*

(94) mL 2bc d M q.

(95) ´ dBlocking Solution in PBS for Im-.

(96) :+

(97) :)*+(, pH e. munoassay, pH )fµ¶Nq·¸¹je. E6. \. U d mLe 2%. 21£Npq¥#º d x cme. NII-Electronic Library Service.

(98) Shiga University.

(99). ®¯. °±/0 ²³±1 ´2±µ3¶±·4 5¸±¹6 º ±»3 7¼. h

(100) . mg ) Y AGLῌ fr. . mL . mg ) KLM fr. mg ).

(101) . Y AGLῌ 5ST&'l ^. . ! "#$ B %& '() . yW4o.4Y@. * h +,-". . ^ ¡W* A** fr. . /0 PBS 123 4 5. A* AGLῌ a KLM fr. A* AGLῌ.

(102) . b 5 fr. mg ) Y. mL 6$789": ;<=. AGLῌ 5 fr. mg ) Y. IgG H>L) Jackson Immuno Re-. AGLῌ 5 AGLῌ ST&5 ¢1. search Lab., Inc.) *. ?@+. KLM¢1$YA** mg . ,-". A0 PBS 123. £ Ws STi5(. B C mg ῌchloroῌ ῌnaphtol . fr. fr. KLM fr. i4Y. mL DEF"$G H mL . _!£w ῌ F"$ / Q / . mM TrisῌHCl IJ pH ) KLM . Q S T ) * ¤ ReῌIatrobeads. mL NOPQRQST) U

(103) V. column. WX /Y Z[V\]^_` 5. §+ fr. zsY mg AGLῌ a. chromatography L}~¥¦. abQ24 c ῎῏ ῎ῐ). fr. zsY mg AGLῌ b fr. z s Y mg AGLῌ , Fig. ῎,. ῏ . lanes ῍ ῑ ). Fig. ῏ -¨©ª+u. ῏. ῍.

(104) Pleurotus cystidiosus. u. «]. P^¬.4w77deEftn. AGLῌ῍, AGLῌ. £wx Pleurotus eryngii 5 TLC. ῐ a, AGLῌ ῐ b, AGLῌ ῑ AGLῌ ῒ. E " ) l ^ p q £ w. 77deEfOghCiY TLC jK4 orcinolῌHSO kKLM Hanesῌ Isherwood kglm n. o.^pq5rW* Fig. ῍, lane ῌ ) a*so.Y tFu5. vW*4wx A*s5y ^mw5abzs AGLῌ KLM AGLῌ 5{| Ws a*spq }~N q5 AGLῌ a KLMῌ b 5 AGLῌ 5{|mw n }~Hq 77deEf OghCi mg) zs Iatrobeads column . 4 ῌ F"$ / . Q!_"L i frs. ) i Fig. ῍, lanes ῍ ῒ ) . iY TLC o.yE" # A** L. fr. KLM1$ mg ). Y%. hC AGLῌ Ins ῌPOῌCer). KLM AGLῌ nST& fr.. Fig. ῍. Fractionation of Mycoglycolipids of Pleurotus cystidiosus Ohiratake by Iatrobeads Column Chromatography. The column was eluted with a ῌpropanol/water/ammonia : : , v/v). Separation of AGL fraction lane , the acidic glycolipids obtained by DEAEῌ Sephadex fractionation procedure) yielded six subfractions designated frs. to lanes to ). The plates were developed in ῌ propanol/water/ammonia : : , v/v) for h, and the spots were visualized with orcinolῌ HSO reagent for Panel A and with HanesῌIsherwood reagent for Panel B.. NII-Electronic Library Service.

(105) Shiga University. ¼῍¦@ 1±².

(106). AGLῌ AGLῌ ΐ

(107). ῌῌ AGLῌ . !"#$. AGLῌ b %&'(

(108) ) *+ ,. - . /0ῑ. !"#$ AGLῌ a. AGLῌ %&'(1234 56 789:;<= >?@A@BC DE FGHI/JK(LMN6OP QR78'(>?@A@BCS'(% Fig. ῍. Thin Layer Chromatograms of the Mycoglycolipids Isolated from P. cystidiosus

(109) Ohiratake . Lane , AGL fraction obtained from DEAEῌSephadex column chromatography ; lanes to , isolated AGLῌ , AGLῌ a, AGLῌ b, AGLῌ and AGLῌ . The plates were developed in ῌpropanol/ water/ammonia : : , v/v for h, and the spots were visualized with orcinolῌHSO reagent for Panel A and with HanesῌIsherwood reagent for Panel B.. TU V῕WH5 T/XY (LM/Z[ N=/6 AGLῌ a ,- \] 0ῑ/. <). DE^῕/_=LMN6 à. bcHI9JdLM/<=NeE fgbchije klm no9Jd MpE ,q r῔ s῕ nt/u῎ *(LM9vw'( 6 AGLῌ AGLῌ 56 IR 1xy4z9. [ML{ ΐ M) cmῌ v|}~C῏'(.=^. / AGLῌ 93b ῐyz / #z ῌ W+ "~C%&'( )n( ῎. ῍. AGLῌ ῌ . AGLῌ ῏ b. AGLῌ ῐ . AGLῌ ῑ . AGLῌ AGLῌ b AGLῌ AGLῌ. M ¡ HI¢9 Table ῌ £ ¤W ¥ ῌ!C ¦§¨© ῌ!C¦§A 9ªWHM «!C¦§ ῌ!C¦§ ¬®¯ῒ ° : ° ¡¤W) ¥ ° Fig. ῎. Thin Layer Chromatogram Comparing the Mycoglycolipid Patterns between P. cystidiosus

(110) Ohiratake and the King Oyster Mushroom, Pleurotus eryngii

(111) Eringi . Lane , AGL fraction from P. cystidiosus Ohiratake ; lane , AGL fraction from P. eryngii Eringi. The plate was developed in ῌpropanol/water/ammonia : : , v/v for h, and the spots were visualized with orcinolῌHSO reagent.. ±41±²§*e L6 K ³("~C¤W ´µ¶(Q R*( >?@A@BC AGLῌ AGLῌ AGLῌ 56. M\· ±41±²§M ῌ!C ¦§9WHM'(±4"~C9¸¹M (LM/Z[ ῌῌ , AGLῌ %&'(>?@A@BC. @}0ῑ º»9Je=. NII-Electronic Library Service.

(112) Shiga University.

(113). . Table ῌ. g { ¡¢ £¤¥ ¦§. Aliphatic Compositions ῌCeramide Compositions῍ of Mycoglycolipids from P. cystidiosus ῌOhiratake῍. AGLῌ AGLῌb AGLῌ AGLῌ Composition Fatty acid IM Nonhydroxy acid : : : : ῌHydroxy acid tr. tr. . . : . : . . : : . : . . : . Sphingoid IM t : t, trihydroxy sphingoid IphytosphingosineM ; tr, trace..

(114)

(115)

(116) ῎. ῎. MALDIῌTOF MS ῏῎. !"#$%#%&'(

(117) )*

(118) "+,./012 3"+,456 7'8 9 AGLῌ AGLῌ b AGLῌ :8; AGLῌ MALDIῌTOF MS <=>?@ ABC D Fig. ῏ ' E6FGHF IJMῌHKLM Table ῍ 'N @ !"#$%#%& O P # Q 5 R S ICer T FA U LCBῌHOM %,V IHPOM "WX5D IInsM :8;!,W5A IManM 899. Y2Z[\] @ ManῌInsῌJPOKῌCer . Fig. ῏. NegativeῌIon Linear Mode MALDIῌ TOF MS Spectra of AGLῌ ῌ, AGLῌ ῏b, AGLῌ ῐ and AGLῌ ῑ. A : AGLῌ ; a, JMῌHKL ion at m/z ; b, m/z ; B : AGLῌ b ; a, m/z ; b, m/z ; C : AGLῌ ; a, m/z ; b, m/z ; D : AGLῌ ; a, m/z ; b, m/z .. Table ῍. MALDIῌTOF MS Analysis of AGLῌ ῌ, AGLῌ ῏b, AGLῌ ῐ and AGLῌ ῑ from P. cystidiosus ῌOhiratake῍. Symbol in Fig. A AGLῌ a b B AGLῌb a b C AGLῌ a b D AGLῌ a b. Fatty acid h : h : h : h : h : h : h : h : . Sphingoid t : t : t : t : t : t : t : t : . Found Calc’d JMῌHKL JMῌHKL. . . .

(119) ^_ CerIFAULCBῌHOMUHPOῌHO UInsῌHOUManῌHOTCerIFAULCBῌMU. } 1' ~s!"#$%#%&<. ῌ U ῌ U ῌ T FA U LCB U. { | '

(120) IManῌInsῌJPOKῌCerM U. à YYb ++cde!". nHexῌnHOT U nῌ n :8; . #$%#%&f] ghbij@8k'l. U nῌ n '8?

(121) N12 @ <{|. mV IFAM :8;nopq ILCBM rs. 's ]

(122) InM . t]uv J ῌhydroxy : IMῌt : . 6 a Y s Y . I MT K J ῌhydroxy : IMῌt. AGLῌ AGLῌ b AGLῌ :8; AGLῌ . : I MT K wxs y?

(123) . ABCD IFig. ῏ M :8;E6FGH. ++cde!"#$%#%&OP#. F ITable ῍ M '

(124)

(125) t AGLῌ . Q5z<<{| IMM :8; b. &5CE6F JMῌHKLTm/z . NII-Electronic Library Service.

(126) Shiga University. mU"

(127) Wto*+.

(128). m/z . Fuc : : N^_ ` M.

(129) m / z m / z. ab h c deJfF 'Yg. AGLῌ. h(Y'ij(hZ[\ GC JK. !"#$"$%&

(130) "'()*+,. 0] Bk?@ myoῌ!UVh(YN^_. -./,012 34 AGLῌ b . /,Q AGLῌ ?@lmn Oῌ'i. 5%(6

(131) 781 9MῌH:. XY. ;. <m/z. m/z . oph(YN AGLῌ b AGLῌ . AGLῌ ?@lmn Oῌ'iXY. o. ,0 . = nῌ n. ph(Y lmn Oῌ'iXY qr6Xh(. < = nῌ n<

(132) >. Y soT Oῌ'iXY tVh(YNu.. ? @ A B C n < Hex ῌ. Fig. ῐ 0 @

(133) L M AGLῌ,. ManῌInsῌPῌCer ,-. AGLῌ . ManῌInsῌ9PO:ῌCer ; AGLῌb, FucGal ῌ. 5%(6

(134) 781 9MῌH:; <m/z. ManῌInsῌ9PO:ῌCer ; AGLῌ, FucGal ῌ. m/z . ManῌInsῌ9PO:ῌCer ; AGLῌ, FucGalῌ. ,0 . = nῌ n. ManῌInsῌ9PO:ῌCer - 8 v w G

(135). < = nῌ n<

(136) >. MALDIῌTOF MS JK

(137) LMNxyz{. ?@ ABC n< HexῌMan. ῎. ῏. ῍

(138) . ῌInsῌ9PO:ῌCer ,-./,012. . AGLῌ 5%(6

(139) 781 9Mῌ. )|

(140) L}~NR./, AGLῌ . H:;<m/z m/z . AGLῌ b AGLῌ AGLῌ

(141) SXY. ,0 . = nῌ n < = nῌ n<

(142) > ? @ A B C n < Hex ῌManῌInsῌ9PO:ῌ Cer ,-./,012. D Table. E,. AGLῌ b AGLῌ AGLῌ.

(143) 9MῌH:ῌ5 FG

(144) HI)JK

(145) LMNOP Q

(146) , ῎. ῏. AGLῌ ῌ AGLῌ ῏ a AGLῌ. ῏ b AGLῌ ῐ AGLῌ ῑ ῎. ῏. ῌ . AGLῌ AGLῌ bAGLῌ . AGLῌ

(147) HI)NR./, STU$VWF Oῌ TMSῌSXY#$"V&Z[\. GC JKN0] . AGLῌ,. Man ;. AGLῌ b,. Man : Gal : Fuc : : ; AGLῌ, Man : Gal : Fuc :. : ; AGLῌ, Man : Gal :. Fig. ῐ. Gas Chromatograms of Alditol Acetate Derivatives derived from AGLῌ ῌ, AGLῌ ῏b, AGLῌ ῐ and AGLῌ ῑ. A, myoῌinositol hexaacetate authentic standard ; B, AGLῌ ; C, AGLῌ b ; D, AGLῌ ; E, AGLῌ ; peak a, inositol hexaacetate ; peak b, mannitol hexaacetate ; peak c, fucitol pentaacetate ; peak d, galactitol hexaacetate.. NII-Electronic Library Service.

(148) Shiga University. ῍ῌ. hi. 7jkl mnjop qrjstuvwjxy z{j|} ~jt . AGLῌ ῏ ῏

(149) ΐῌ Oῌ. Oῌ ῏

(150) ῐGal. ῌῐ

(151) ῑ

(152) ῒ

(153) ῔ῌ Oῌ . !"#῏

(154) ΐ

(155) ῔ῌ Oῌῌῐ

(156) ῑ

(157) ῒῌ. ῏ Man

(158) AGLῌ ῒ b . Oῌ ῏

(159) ῔Gal :;<. ῏

(160) ΐῌ Oῌῌῐ

(161) ῑ

(162) ῒῌ Oῌ. =>(? @AB Fig. ῏ 9> 4. ῏ Fuc

(163) ῏

(164) ΐῌ Oῌῌ. 9: C4"DEFG5HIJ1K<8L. ῐ

(165) ῑ

(166) ῒ

(167) ῔ῌ Oῌ . . AGLῌ ῏ Ins M Man 15HBN)( AGLῌ. ῏ Gal

(168) ῏

(169) ΐ

(170) ῔ῌ Oῌῌῐ

(171) ῑ

(172) ῒῌ. ῒ b AGLῌ ῏ 4 Man 4 ῔ IM ῐ I: ῔ I8. Oῌ ῏

(173) ῔Man !". OB Gal 15HB

(174) = MP4O4). #῏

(175) ῐ

(176) ΐ

(177) ῔ῌ Oῌῌῑ

(178) ῒῌ Oῌ. Q> M Fuc : Gal 1P>R> ῏ $QS5. ῏

(179) ῐ

(180) ῔Gal ῌῌ

(181). H B N ) ( AGLῌ ΐ AGLῌ ῒ b 4 Man M. AGLῌ ΐ ῏ Fuc

(182) ῏ Gal ῐ $

(183) ῏

(184) ῔. 5HBN)( ῐ I!"# ῔ I84O Gal M. Man !"#῏

(185) ῐ

(186) ῑ

(187) ΐ

(188) ῔ῌ%& Oῌῌ. = M ῑ IM Gal 15HBTOUVW. ῒῌOῌ ῏

(189) ῐ

(190) ῑ

(191) ῔Gal. BN)(X48'( AGLῌ ῔ Man : Fuc :. '() ῏

(192) ῐ

(193) ῒ

(194) ΐ

(195) ῔ῌ%& Oῌῌῑῌ. Gal 4$Y1

(196) !!"P ῏ : ῏ : ῒ 8'(9. Oῌ *+ ,-.. :

(197) !"#῏

(198) ῐ Gal :῏

(199) ῔ Gal 4$Y1 ῎῕ῒ :. .% /01

(200) 234 AGLῌ ΐ 4. ῎῕ΐ 8'(9:

(201) AGLῌ ΐ 4Z[\]^M. 56 7+8'(9:1;<=>( ῌῌ. IJ0( ῐ S4 Gal 4D_4`_ M

(202) =.

(203) AGLῌ ῔ ῏ Fuc

(204) ῏ Gal

(205) ῏

(206) ῔Man

(207). M ῏ ῌ ῐ 8 Gal 15HBX4: ῏ ῌ ῔ 8 Gal. ῏

(208) ῐ

(209) ῑ

(210) ῔ Gal

(211) ῏

(212) ῐ

(213) ΐῌ Oῌῌῑ

(214) ῒ

(215) ῔ῌ. 15HBX44 ῐ ab1cd0(efg1/. Fig. ῏. Gas Chromatograms of Partially Methylated Alditol Acetate Derivatives derived from AGLῌ ῌ, AGLῌ ῍b, AGLῌ ῎ and AGLῌ ῏. A, AGLῌ ῏ ; B, AGLῌ ῒb ; C, AGLῌ ΐ ; D, AGLῌ ῔ ; peak a, ῏,ΐ ῌdiῌOῌacetylῌ ῐ,ῑ,ῒ,῔ ῌtetraῌOῌ methylmannitol ῏Man ; peak b, ῏,ΐ ῌdiῌOῌacetylῌ ῐ,ῑ,ῒ ῌtriῌOῌmethylfucitol ῏Fuc ; peak c, ῏,ΐ ῌ diῌOῌacetylῌ ῐ,ῑ,ῒ,῔ ῌtetraῌOῌmethylgalactitol ῏Gal ; peak d, ῏,ΐ,῔ ῌtriῌOῌacetylῌ ῐ,ῑ,ῒ ῌtriῌOῌ methylmannitol ῏,῔Man ; peak e, ῏,ῐ,ΐ,῔ ῌtetraῌOῌacetylῌ ῑ,ῒ ῌdi ῌOῌmethylgalactitol ῏,ῐ,῔Gal ; peak f, ῏,ῐ,ῑ,ΐ,῔ ῌpentaῌOῌacetylῌ ῒ ῌOῌmethylgalactitol ῏,ῐ,ῑ,῔Gal or ῏,ῐ,ῒ,ΐ,῔ ῌpentaῌOῌacetylῌ ῑ ῌ Oῌmethylgalactitol ῏,ῐ,ῒ,῔Gal ; peak g, ῏,ῐ,ΐ ῌtriῌOῌacetylῌ ῑ,ῒ,῔ ῌtriῌOῌmethylgalactitol ῏,ῐGal ; h, ῏,ΐ,῔ ῌtriῌOῌacetylῌ ῐ,ῑ,ῒ ῌtriῌOῌmethylgalactitol ῏,῔Gal. Under these analytical conditions, all the expected inositol products remain phosphorylated and are hence not observable.. NII-Electronic Library Service.

(216) Shiga University. µ¶©.·¸¹º»¼. .

(217)

(218). M 5 !"5 &=.

(219) . u'y;3$(J "y. ! "ῌῌ# $. AGLῌ K'<)5 wxyz{|<*. %&''()*+,-./.01. }# yz{|3&5 AGLῌ. !$23435 6789. TLC =+. <>5 AGLῌ. :;3<=> 5 AGLῌ ?65. " #5 AGLῌ " #5 AGLῌ " a#5 AGLῌ. @ A=65 BC&

(220) DE F Gal. " b#5 AGLῌ " # 3& "Fig. ῒ #. GHIJ ,-./. 7,-$R: mg AGLῌ. .01<KL:MN<%OP. $ .o mg ?¡. AGLῌῌ, Man ῌ Ins ;. &5 R=wxy$~J5 /x. AGLῌ ῏ b, Gal ῌ ῌ ῑ Fuc ῌ ῌ ῍ Gal ῌ ῌ ῑ. 0=1 " h# & 1 23¢$£¤¥. Man ῌ Ins ;. &5 mg yz{|$ ;$ ῌ. AGLῌῐ, Gal ῌ ῌ ῑ Gal ῌ ῌ ῎ Fuc ῌ ῌ ῍ . ¦§¨©[ / x / Xª«Xx " : : #. Gal ῌ ῌ ῑ Man ῌ Ins ;. ¬ 4 4 5 Iatrobeads column. AGLῌῑ, Gal ῌ ῌ ῍ / ῑ Gal ῌ ῌ ῑ Gal ῌ ῌ ῎ . chromatography "column sizef ® cm5. Fuc ῌ ῌ ῍ Gal ῌ ῌ ῑ Man ῌ Ins ;. bed volume mL "f ® cm#5 collec-. Gal ῌ ῌ ῑ Gal ῌ ῌ ῍ / ῑ Gal ῌ ῌ ῎ . tion volume mL/tube# =¯5 }&. Fuc ῌ ῌ ῍ Gal ῌ ῌ ῑ Man ῌ Ins.. "/°$ mL/min 3:45 w¤±²$. ῎. ῏. ῎ AGLῌ ῐ

(221) . ~ J # ³ ´ 6 AGLῌ " #5 mg ;. . AGLῌ " #5 mg ; AGLῌ " a#5 mg ;. . AGLῌ " b#5 mg ; AGLῌ " #5 mg. !". % &'()*. #$ +,-./. 0 QR&85 STU. =J oyz{|$ z& 0Nz65 AGLῌ " # < Man5 AGLῌ. !VW$&. " # < Man : Gal " : 5 ª #5 AGLῌ. AGLῌ b5 AGLῌ 5 AGLῌ "intact AGLῌ. b, intact AGLῌ, intact AGLῌ # .

(222) X YZ[ X\][Z ^P65 _ à Gal bGHc Fuc GH$ d:;36=>U efU;35 g hijUkl ,-./.01 !<m ;3^P5 Qn=6opq5 !$rsRt<5 ;P$ STUu3:4v_P AGLῌ $5 owxyz{|$. }&y $~J 5 FucῌGal G$y&5 5 < "Galῌ # "Galῌ # Galῌ Man = u ";$ AGLῌ " # 3:# $ yz{|^P5 }&

(223) X YZ[ X\][Z $~J ;wxy$d:4 U3&5 3!" "# #$$& TLC %=65. Fig. ῒ. Thin Layer Chromatogram of Partial Acid Hydrolyzates of AGLῌ ῐ. Lane , intact AGLῌ ; lane , acidic hydrolyzates of AGLῌ ; lanes to , purified AGLῌ "#, AGLῌ " #, AGLῌ "a#, AGLῌ "b# and AGLῌ "#. The plate was developed in ῌpropanol/water/ ammonia " : : , v/v# for h, and the spots were visualized with orcinolῌ H SO reagent.. NII-Electronic Library Service.

(224) Shiga University. ῌ῍. JK. LMNO PQMRS TUMVWXYZM[\ ]^M_` abMcW de. ῒ ῐ a AGLῌ ῒ ῐ b Man :. 9:3 BCD2E3 ῎ Man AGLῌ ῒ. Gal ῎ : ῏ AGLῌ ῒ ῑ Man : Gal ῎ :. ῏ )῎ ῒῌ= Oῌ>?:3ῌ῏ ῐ ῑ ΐῌ@2. ῐ

(225) intactῌ. A Oῌ9:3 FA1:2E3 ῎ Gal . AGLῌ ῒ Fuc. ῎ ῒ ΐῌ2G Oῌ>?:3ῌ῏ ῐ ῑῌ2G Oῌ. . 9:3 BCD2E3 ῎ ΐMan AGLῌ. !"#$% $ &' MALDIῌ. TOF MS ("') AGLῌ ῒ. ῎ ) Manῌ. ῒ ῐ a ) ῎ Gal ῎ ΐ Man ῎ ῐ ῒῌ. Insῌ*POῑ+ῌCer AGLῌ ῒ ῏ ) GalῌManῌ. 2G Oῌ>?:3ῌ῏ ῑ ΐῌ2G Oῌ9:3. Insῌ*POῑ+ῌCer AGLῌ ῒ ῐ a AGLῌ. F A 1 : 2 E 3 ῎ ῐ Gal AGLῌ ῒ. ῒ. ῐ b ) ῎ Gal ῎ ΐMan ῎ ῒ ΐῌ2G. ῐ b ) GalῌGalῌManῌInsῌ*POῑ+ῌCer. AGLῌ ῒ ῑ ) Gal῏ ῌGalῌManῌInsῌ*POῑ+ῌ. Oῌ>?:3ῌ῏ ῐ ῑῌ2G Oῌ9:3 FA1. Cer

(226) &,- ./01234'. :2E3 ῎ ΐGal AGLῌ ῒ ῑ ). $ Data not shown% 56 7'8. ῎ Gal ῏ H3 ῎ ΐMan ῎ ῐ ῒ ΐῌ@. 9:3;("' 7< AGLῌ ῒ ῎ . 2A Oῌ>?:3ῌ῏ ῑῌ= Oῌ9:3 FA. )῎ ῒῌ= Oῌ>?:3ῌ῏ ῐ ῑ ΐῌ@2A Oῌ. 1:2E3 ῎ ῐ ΐGal I-$ Fig. ῍ %. Fig. ῍. Gas Chromatograms of Partially Methylated Alditol Acetate Derivatives derived from Acid Hydrolyzates of AGLῌ ῌ. A, AGLῌ ῒ῎ ; B, AGLῌ ῒ῏ ; C, AGLῌ ῒῐa ; D, AGLῌ ῒῐb ; E, AGLῌ ῒῑ ; peak a, ῎,ῒ ῌdiῌOῌ acetylῌ ῏,ῐ,ῑ,ΐ ῌtetraῌOῌmethylmannitol ῎Man ; peak b, ῎,ῒ ῌdiῌOῌacetylῌ ῏,ῐ,ῑ,ΐ ῌtetraῌOῌmethylgalactitol ῎Gal ; peak c, ῎,ῒ,ΐ ῌtriῌOῌacetylῌ ῏,ῐ,ῑ ῌtriῌOῌmethylmannitol ῎,ΐMan ; peak d, ῎,ῐ,ῒ ῌ triῌOῌacetylῌ ῏,ῑ,ΐ ῌtriῌOῌmethylgalactitol ῎,ῐGal ; peak e, ῎,ῒ,ΐ ῌtriῌOῌacetylῌ ῏,ῐ,ῑ ῌtriῌOῌmethylgalactitol ῎,ΐGal ; peak f, ῎,ῐ,ῒ,ΐ ῌtetraῌOῌacetylῌ ῏,ῑ ῌdiῌOῌmethylgalactitol ῎,ῐ,ΐGal. Under these analytical conditions, all the expected inositol products remain phosphorylated and are hence not observable.. NII-Electronic Library Service.

(227) Shiga University. [Ù6TL.

(228) .

(229) AGLῌ . AGLῌ 6~34)6 +WMῌHX : m/z. . . PQ m/z , +Fig. ῐ ῌ ῍ A, ῍. !" #. B, ]C 6. ?

(230) @#. AGLῌ ῑ ῍ Man ῍ ῌ ῎ Ins ῍ ῌPOῐῌCer ;. 6. AGLῌ ῑ ῎ Gal ῍ ῌ ῒ Man ῍ ῌ ῎ Ins ῍ ῌ. u]@ WMῌHX m/z ῌ PQ. ῍+I,ῌ+III, ;. POῐῌCer ;. WMῌHX m/z ῌ ]CB. AGLῌ ῑ ῏ a Gal ῍ ῌ ῏ Gal ῍ ῌ ῒ Man ῍ ῌ ῎. -D Man. Ins 8: B 8. Ins ῍ ῌPOῐῌCer ; AGLῌ ῑ ῏ b Gal ῍ ῌ ῒ Gal ῍ ῌ ῒ Man ῍ ῌ ῎ Ins ῍ ῌPOῐῌCer ; AGLῌ ῑ ῐ Gal ῍ ῌ ῒ Gal ῍ ῌ ῏ Gal. ῍ ῌ ῒ Man ῍ ῌ ῎ Ins ῍ ῌPOῐῌCer. $ %& AGLῌ intactῌAGLῌ '()* +Fig. ῒ ῌC, - ./0123456 789 8!:; <=7 +Gal ῌ , +Gal ῌ , +Fuc ῌ , Gal ῌ. Man ῌInsῌPῌCer !: . >?$@#. @ AGLῌ b ABCD D E7 F- AGLῌ GHI;J KL MN O intactῌAGLῌ b +Fig. ῒ ῌ B, P Q AGLῌ + , + . RCSTU56$CB , '() * +Fig. ΐ ῌE, - V<=7 +Gal ῌ , +Fuc ῌ , Gal ῌ Man ῌ InsῌWPOXῌCer !: 7YZC>?$@#. ῏. ῐ. ῐ .

(231) ManIns Man Ins 789ABC AGLῌ. [BC Fig. ῔ \]@^_6`5a RCJ bcdeff*;7 89 gh@# ij Table ῏ \] @^_kl!: <m)U5) n;V789 o- pq !: AGLῌ . + ῌ , 7!:r. kl<m)U5) ]Cst4t45 )uv >?!" I # Fig. ῍ῌ ^_6`5a +Fig. ῔ , +III, wx ^_yz +bcdeff*^_{|} , MALDIῌTOF MS 6~34) \ ]@# 6~34)- WMῌHX : m/z m/z +Fig. ῍ῌ ῌa, b, u]@ BD῍WMῌHX : m/z PQ . m/z ]@# @ intactῌ. Fig. ῔. Proposed Reaction Schemes of the Periodate Oxidation of AGLῌ ῍I, and Subsequent Processing Leading to Erythritol V.. NII-Electronic Library Service.

(232) Shiga University. ῌ῍. xy. Table ῏. z{|} ~{ { ^{ { { . Possible Substitutions on myoῌ Inositols by Mannose in AGLῌ ῍. Possible substituted positions on myoῌinositols of AGLῌ ῎* ῎,῏ ῎,ῐ ῎,ῑ ῎,ῒ ῎,ΐ. Final alchohl products derived from substituted myoῌinositol** erythritol ribitol glycerol xylitol threitol. *Phosphate group is linked to myoῌinositol at the Cῌ ῎ position. **The predicted corresponding final myoῌinositolῌ derived alcohol products after periodate oxidation followed by NaBHῑ reduction, methanolysis, enzymatic dephosphorylation and acetylation.. Fig. ῍῍ A Final Acetylated Alcohol Product derived from the myoῌInositol Ring of AGLῌ ῍ by Periodate Oxidation. A, authentic erythritol EaF and threitol EbF peracetates ; B, erythritol peracetate EcF derived from the myoῌinositol ring in AGLῌ῎. *byῌproduct. Fig. ῍ῌ NegativeῌIon Linear Mode MALDIῌ TOF MS Spectrum of a Periodate Oxidation Product SchemeῌIII in Fig. ῒ of AGLῌ ῍.. $H* Man# aῌ$H* Gal 567 bῌ$.

(233) . Gal # AGLῌ ῒ E ῑ F < A aῌ$ H *. !"# $%. Man# aῌ$H* Gal E ῏ ]%8F 567 bῌ. &. $H* Gal # AGLῌ ῑ b <A aῌ$H*. '"()*+,-./$%. H * Gal # AGLῌ ῒ E ῐ bF < A aῌ$ H* Man# aῌ$H* Gal 567 bῌ$H*. 0*%$12%34# GC 567 GCῌMS 8. Man# aῌ$H* Fuc# aῌ$H* Gal 56. 9, :;# AGLῌ ῎ <= >. 7 bῌ$H* Gal # AGLῌ ῒ <A aῌ$. >*%?@ < Man A Ins ῏ B. H* Man# aῌ$H* Fuc# aῌ$H* Gal. CD EFig. ῍῍F. E ῏ ]%8F 567 bῌ$H* Gal # AGLῌ. ῏. ῐ. ῑ. ῍. HῌNMR

(234) . . ΐ <A aῌ$H* Man# aῌ$H* Fuc# aῌ$ H * Gal E ῐ ] % 8F 5 6 7 bῌ$ . /G$H*IJCDKL AGLῌ. H* Gal ^_K![`% :abL. ῎ # AGLῌ ῑ b# AGLῌ ῒ # AGLῌ ΐ 567 AGLῌ. ;<cdefgh. ῒ M8NO8P EAGLῌ ῒ E ῐ aF#. i A# AGLῌ ῒ E ῐ aF 567 AGLῌ ῒ E ῐ bF. AGLῌ ῒ E ῐ bF# AGLῌ ῒ E ῑ FF Q῎ Hῌ. 5j:a ῏ Q Gal Eklmn Gal. NMR 89RS Fig. ῍῎ :"T. 567oklmn GalF $H*IJpq. U>%# V Table ῐ 3W!(>567. $H*IJ rDKs# t. &XY Z[D\ # AGLῌ ῎ <A. ui AGLῌ ῒ v AGLῌ ΐ 5jw\ Gal . aῌ$H* Man # AGLῌ ῒ E ῐ aF <A aῌ. p ῎ Q<i bῌ$H* Gal CDK. NII-Electronic Library Service.

(235) Shiga University. §¨;OU©Lª«¬.

(236). Gal _` 6789 Gal @ aῌ:;<=. 56789 Gal @ bῌ:;<=nBC. AGLῌ AGLῌ a ,2Z AGLῌ. b op@"qr AGLῌ k567 89 Gal st bῌ:;<= uv$2_ AGLῌ :;<= >? ^Y aῌ:;<= Man ^Y aῌ:;<= Gal ^Y bῌ:;<= Gal . 23 ^Y bῌ:;<= Gal w xyBI z!" AGLῌ a , 2Z AGLῌ b k Gal _` 6 789 Gal bῌ:;<= 56789 Gal aῌ:;<=E{ |` AGLῌ. AGLῌ b AGLῌ ,2Z AGLῌ ,"ta&b Gal c bῌ:;<= }B$ I #)4I GHI" ~;O <NOPKOKQR:; Fig. ῌ῍ Anomeric Proton Regions of the ῌHῌ NMR Spectra. A : a, AGLῌ ; b, AGLῌ b ; c, AGLῌ ; d, AGLῌ ; I, ManaHῌ ; II, GalbHῌ ; III, FucaHῌ , IV, GalaHῌ ; V, Gala Hῌ ; VI, GalaHῌ ; B : a, AGLῌ a, b, AGLῌ b ; c, AGLῌ ; I, ManaHῌ ; II, GalbHῌ ; III, GalaHῌ ; IV, GalaHῌ . *FucaHῌ.. <=>?C AGLῌ Mana ῌ. AGLῌ Galb ῌ Mana ῌ Gala ῌ . X ppm @"l C AGLῌ a ,2Z AGLῌ b opt aῌ:;<= Man aῌ:;<= Gal ,2Z bῌ:;<= Gal PYl. I E? ῌ ,2Z ῌ 2qrI Gal :;<=.

(237) . X$ AGLῌ . AGLῌ a AGLῌ b . , " AGLῌ a AGLῌ . nativeῌAGLῌ . b op XC P. !" #$ AGLῌ %&'(&. Y$ IntactῌAGLῌ ,2Z intactῌ. )*+,- AGLῌ ./01. AGLῌ ," FucῌC P. 23 NMR &4 AGLῌ . Y ppm lIt aῌ:;. AGLῌ 56789 Gal :;<=>. <= Fuc bῌ:;<= Gal EC. ?@ABC D&4EF. 2!"56789 aῌ:;<= Gal ῌ aῌ. GHI"JKLM<NOPKOKQR. :;<= Man XC. ST7 NMR UVWXY AGLῌ ,2Z. $ #$ AGLῌ ,". AGLῌ [#\] ^Y,2Z ^. 56789 Gal aῌ:;<= ῌ ,2Z. Y Gal _ ` a & b Gal c bῌ: ;. ῌ E" Gal P. <=deI ffghij. Y op@" ^Y&C2_. AGLῌ a AGLῌ b ,2Z AGLῌ. ppm ppm ¡$ . NMR UVWXY \k. AGLῌ 56789 Gal ¢ E@£. "l@mc. Ct ¢ E@£Ct¤. AGLῌ a ,2Z AGLῌ b k. @de",3 v¥¦Y&4. NII-Electronic Library Service.

(238) Shiga University. .

(239) . Table ῏. q * . Chemical Shifts and Jῌ,῍ Coupling Constants of the Protons of AGLῌ ῌ, AGLῌ ῏b, AGLῌ ῐ, AGLῌ ῑ, AGLῌ ῐ῎a, AGLῌ ῐ῎b and AGLῌ ῐ῏ in the Anomeric Regions. AGLῌ Chemical shifts ppm Coupling constants Hz AGLῌ b Chemical shifts ppm Coupling constants Hz AGLῌ Chemical shifts ppm Coupling constants Hz AGLῌ Chemical shifts ppm Coupling constants Hz AGLῌ a Chemical shifts ppm Coupling constants Hz AGLῌ b Chemical shifts ppm Coupling constants Hz AGLῌ Chemical shifts ppm Coupling constants Hz. ManaῌInsῌPῌCer ῌ GalaῌFucalῌGalbῌManaῌInsῌPῌCer ῏ ῎ ῍ ῌ . . GalaῌGalaῌFucaῌGalbῌManaῌInsῌPῌCer ῐ ῏ ῎ ῍ ῌ . . . Galaῌ/GalaῌGalaῌFucaῌGalbῌManaῌInsῌPῌCer ῑ ῐ ῏ ῎ ῍ ῌ or or GalaῌGalbῌManaῌInsῌPῌCer ῎ ῍ ῌ . GalaῌGalbῌManaῌInsῌPῌCer ῎ ῍ ῌ GalaῌGalaῌGalbῌManaῌInsῌPῌCer ῏ ῎ ῍ ῌ . ῎. ῏. ῍

(240) . G]^_à,b% Gal %g'J. ῎. ῏. ῑ.

(241) . hiE)@LMC

(242) j k'( lm. . G "AGLῌ a Gal ῌ Fuc ῌ . AGLῌ AGLῌ . . ! AGLῌ Gal ῌ. Gal ῌ Man ῌ Ins ῌ"PO#ῌCer# "AGLῌ b Gal ῌ Fuc ῌ . Gal ῌ Fuc ῌ Gal ῌ Man ῌ. Gal ῌ Man ῌ Ins ῌ"PO#ῌCer#

(243) . Ins ῌ"PO#ῌCer $% &'( )*. nF op

(244) TLC N

(245) OPQ [\%iG. +,-. /0123456,7% aῌ. qF

(246) &E AGLῌ r)>?)@U. bῌ/012893:, ;<=>?)@. VW AGLῌ a AGLῌ b. ABC TLC Fig. ῌ῎ ,DCE. OPQ Fig. ῒ ῌlanes ῐ ῑ sRJ(. F aῌ/012893:

(247) G)@H'C. tu

(248) \C hi vvw0xy AGLῌ . E bῌ/012893:

(249) G)@H'IJK. aῌ/012893:

(250) B )@

(251) TLC N. C AGLῌ G aῌ/012893. ,NzC AGLῌ )@UVW. :

(252) )@LM TLC N

(253) OPQ RI. {Y|PD ST 4}~2Eu

(254) \C. ST )@UVWXYC Fig. ῌ῎ ῌlane. Fig. ῌ῎ lane ῐ k'(Gk'' AGLῌ. ῍ OPQ Z [\%UVWG]^_à. Fuc ῌ Gal ῌ Man ῌ Ins. ,b% cd Gal Ee)@H. ῌ"PO#ῌCer ; AGLῌ a Gal ῌ . 'C "AGLῌ Fuc ῌ Gal ῌ . Fuc ῌ Gal ῌ Man ῌ Ins ῌ"PO#ῌ. Man ῌ Ins ῌ"PO#ῌCer#

(255) f ST. Cer ; AGLῌ b Gal ῌ Fuc ῌ . NII-Electronic Library Service.

(256) Shiga University. lJm .

(257). Fig. ῌ῍ Thin Layer Chromatogram of the Reaction Products from the Mycoglycolipids, AGLῌ ῏ and AGLῌ ῐ obtained by Treatment with aῌand bῌGalactosidases. Lane , AGLῌ /from Eringi, P. eryngii0 ; lane , AGLῌ /from Eringi0 incubated with aῌgalactosidase ; lane , AGLῌ /from Eringi0 incubated with bῌgalactosidase ; lane , AGLῌ /from Ohiratake, P. cystidiosus0 ; lane , AGLῌ /Ohiratake0 incubated with aῌgalactosidase ; lane , AGLῌ /from Ohiratake0 incubated with bῌgalactosidase ; lane , AGLῌ /from Ohiratake0 ; lane , AGLῌ /ohiratake0 incubated with aῌgalactosidase and lane , AGLῌ /Ohiratake0 incubated with bῌgalactosidase. The plate was developed in ῌpropanol/water/ammonia / : : , v/v0 for h, and the spots were visualized with orcinolῌHSO.. Gal ῌ Man ῌ Ins ῌPOῌCer . AGLῌ

(258) aῌ

(259) TLC . ῍. ῎. ῑ B TLCῌ immunostaining. !". bcd[ef#M:ghij O [ /H. #$% &'()* + ,-.-. [0 Fuca ῌ Galb k ; A [ GalNAca ῌ. AGLῌ / 0 /Fuc ῌ 0 Gal ῌ Man ῌ . Fuca ῌ 0 Galb k ; B [ Gala ῌ . Ins ῌPOῌCer ; AGLῌ / a0 /Gal ῌ 0. /Fuca ῌ 0 Galb k7L; ?WXlJ. /Fuc ῌ 0 Gal ῌ Man ῌ Ins ῌPOῌ. m 2

(260) nopq Hypsizigus marmo-. Cer ; AGLῌ / b0 /Gal ῌ 0 /Fuc ῌ 0. reus Krmstmtuv AGLῌ /Gala ῌ. Gal ῌ Man ῌ Ins ῌPOῌCer ;. /Fuca ῌ 0 Galb ῌ Mana ῌ Ins ῌ. AGLῌ. / 0 /Gal ῌ 0 /Gal ῌ 0 /Fuc ῌ 0. PO ῌCer0 BCDEF5?- 89:;. Gal ῌ Man ῌ Ins ῌPOῌCer . gh#w

(261) ; / 0 xy zzb{|. /Fig. ῌ῍ lane ῒ 0 1 AGLῌ 234. AGLῌ b BCDEF}gh Gala ῌ. AGLῌ 5 6 aῌ 7. /Fuca ῌ 0 Galb k AGLῌ BCD. AGLῌ / 0 /AGLῌ / 0 89:;. EF}gh YZ[ Gal G234 . #<=> ?@A AGLῌ / 0 /. G Gal 5 P ~ /Gala ῌ 0 /Gala ῌ. AGLῌ / 0 BCDEF GH:; Gal . 0 /Fuca ῌ 0 Galb k AGLῌ OA. bῌIJK7L;?5MNO-. aῌGal 5 AGLῌ BCDEF} Gal . ?-APQ@A AGLῌ 234 AGLῌ . GL; G7P~

(262) 2] -b. RS T1-; UVL; UV. B [cdef#w:;f#T7 ;. . AGLῌ 234 AGLῌ

(263) . WX YZ[ Gal \5 bῌI. JK7L] ^] _

(264) aῌIJK7L;. BCDEFgh b B cd[ef#. ?5à. w : ; g h i j Gala ῌ /Fuca ῌ 0 Galb k#

(265) ; /Fig. ῌ῎0 1 . NII-Electronic Library Service.

(266) Shiga University. ῌ῍. . 8 ` ¡¢£ ¤¥¦ §¨. Fig. ῌ῏ Detection of the Mycoglycolipids having a Human Blood Type B Activity by TLCῌImmunostaining. Lane ῏, AGL fraction of P. cystidiosus 4Ohiratake5 ; lane ῐ, AGLῌ ῒa ; lane ῑ, AGLῌ ῒb ; lane ῒ, AGLῌ ΐ and lane ΐ, AGLῌ ῔. The plates were developed with ῏ ῌpropanol/water/ammonia 4῕ΐ : ῑ῎ : ΐ, v/v5 for ῐ h. The spots were visualized with orcinolῌHῐSOῒ 4A5 and immunostaining by antiῌhuman blood type B antibody 4B5.. _

(267) Eà Gala ῏ ῌ ῑ 4Fuca ῏ ῌ ῐ 5 Fig. ῌ῎ Carbohydrate Structures of Type B in Human Blood Group Substances.. Galb ῏ b/0 c

(268) 3. dXSTU B VWAS: 4efgh ijS:5 %klPmn>^ op Gala ῏ ῌ ῑ.

(269) AGLῌ ῒ. 4Fuca ῏ ῌ ῐ 5 Galb ῏ b /0]qL> . a . AGLῌ ῒ b % Gala ῏ ῌ ῔ 4Fuca ῏ ῌ ῐ 5 Galb ῏ b. !"

(270) #$%# &' (. /0CDBr%st *]qL>. ῌ )*+ ῍ % ῐ %,- TLCῌim-. p#M 3 u AGLῌ ΐ à. munostaining * .%/0 12. Gala ῏ ῌ ῑ 4Fuca ῏ ῌ ῐ 5 Galb ῏ b/0 c.

(271) 3.

(272) ]qL>

(273) QR> 3 L. ῌ. AGLῌ ῒ b 4Gala ῏ ῌ ῔ 5 4Fuca ῏ ῌ ῐ 5. AGLῌ ῔ ; ]qL>. Galb ῏ ῌ ῔ Mana ῏ ῌ ῐ Ins ῏ ῌ6POῒ7ῌCer. *"#M>^ à Gala ῏ ῌ ῑ 4Fuca ῏ ῌ. AGLῌ ΐ 4Gala ῏ ῌ ῑ 5 4Gala ῏ ῌ ῔ 5 4Fuca. ῐ 5 Galb ῏ b /0v%wx#HI. ῏ ῌ ῐ 5Galb ῏ ῌ ῔ Mana ῏ ῌ ῐ Ins ῏ ῌ6POῒ7ῌ.

(274) yzL>

(275)

(276) E ?@A bῌGal %. Cer % ῏ %89:

(277) ;<

(278) AGLῌ. ῔ B aῌGal CD .% Gal % ῐ BM. ΐ => ?@A Gal % ῑ B Gal. ῔ B aῌGal CD

(279) 3. C D E " ῏ % 8 9 : F G. AGLῌ ῒ a AGLῌ ῒ b AGLῌ ΐ AGLῌ ῔. 4Gala ῏ ῌ ῑ 5 4Fuca ῏ ῌ ῐ 5 Galb ῏ ῌ ῔. S{

(280) STU B VWAS: * TLCῌ. Mana ῏ ῌ ῐ. ῍. Ins ῏ ῌ6POῒ7ῌCer %HIJ. immuno. staining. !C|. Fig. ῌ῏ . KL> 3 FG > AGLῌ ῒ a #M. (3 AGLῌ ῒ a AGLῌ ΐ AGLῌ ῔ % ῑ }. NOPQR> 3. 4Lanesῌ῍ῌ῎ῌ῏5 [P (

(281) ,~L>. AGLῌ ῒ a STU B VWAS:. X. 8; y

(282) #$3 k . TLCῌimmunostaining *.%YZ. d T *. [P

(283) \]L>>^ AGLῌ ῒ a . %ff AGLῌ ῒ a %/0. NII-Electronic Library Service.

(284) Shiga University. ()*+,WwxE¹/0. ῌ῍. B

(285) . . Gala ῏ ῌ ῑ Fuca ῏ ῌ ῐ Galb ῏ ῌ ῔ Mana ῏ ῌ. "UT%& 'm'U: AGLῌ ῏ IM%]. ῐ Ins ῏ ῌPOῒῌCer . 3^,_' AGLῌ ῎ Ins ῏ ῌPOῒῌCer \. . g: , TLC hi6 ,jkl. no cg& ' %: p U : qgP& ')*+, AGLῌ. ῎. ῐ.

(286) . ῏ ,/ -rs,t: N,)*+,u. . vwxyzUl" '\{|

(287). !"#$%& '()*+,-./. }\ ῍ῐ. ~ : ]3^,_'WXY. 01 TLC ,2345 67 '. Z' : ]3Û: ':. 6῍89+ Pholiota nameko: ;8<9. g: V~I U: N&O. =: >>?@AB: CDEF: C*)AB Fla-. &,KG+`D+Dab, d.. mmulina velutipes: <GAB Lentinus edodes. ' mH: 89+: C*)ABHI. HIJKGAB Grifola. JKGAB,KG+`D+Dab' . frondosa , ῕ L. M: N&O&,-./01 DEAEῌ.

(288) . Sephadex column chromatography PQ. . Fig. ῌῑ N,RS TU: TLC . ῏ . V,-./01UWXYZ%& ' [\]3^,_'WXYZ AGLῌ ῏ . >>?@AB Pleurotus cystidiosus . U: ,KG+`D+Dabc,de. ΐ L,KG+`D+Dab,. \,, ῕ L ,)*+f '. ,I{. Fig. ῌῑ Thin Layer Chromatogram Comparing the Mycoglycolipid Patterns among Seven Fungi. Lane ῏, Pholiota nameko Nameko ; lane ῐ, Hypsizigus marmoreus Bunashimeji ; lane ῑ, Pleurotus cystidiosus Ohiratake ; lane ῒ, Pleurotus eryngii Eringi ; lane ΐ, Flammulina velutipes Enokitake ; lane ῔, Lentinus edodes Shiitake and lane ῕, Grifola frondosa Maitake. The plate was developed in ῏ ῌpropanol/water/ ammonia ῕ΐ : ῑ῎ : ΐ, v/v for ῐ h, and the spots were visualized with orcinolῌ HῐSOῒ reagent.. U s ῏ΐ ῏῔ ^ ῐ῎῎ῑ ῐ῎῎ῒ ,+Ey< 1¡: ¢£¤¥_,ῌ¦§H IJ¨©¦§IM bῌ`zªE,«.¬ ,®')*+ >>?@ABU¯°%&: g: ±²,¥®Ud': N, KG+`D+Dab,UWAyZ ῍ῑ. ³´ ῏῎῎ g ,>>?@AB µ¶´. ῏ῑ g ·i%&¸WwxE¹/01 ῏ΐ῎ mg º^: ,1: »@¼b:. NII-Electronic Library Service.

(289) Shiga University. ®. W¯ ! °±¯²" ³#¯´$%µ¶¯·& '¸¯¹ º(¯»$ )¼.

(290)

(291) . eryngii '()* P. ostreatus $)* Cal-.

(292)

(293) . vatia exipuliformis. !"#$

(294) % &. virosa !7}"&&'(). )* Amantia. &'()*

(295)

(296) +, -. * P. cystidiosus "

(297). ."/ 0 1#$

(298) 23.

(299) kYZ76 9. 45,6 789 Fig. ῌ῎ 7:;!. 5 Fuca ῌ Galb ,'. <=>?5@#$

(300) 7A !. Sw O wYZB@ .

(301)

(302) TLC BC@" 4. &&'()* Gala ῌ GFuca. !7DE9F GAGLῌ 5H9I 2J. ῌ I Galb , B wYZB@. 7 K LM ,NM "

(303) O. Type ῍ Gal/Fuc/Man xvw, )*. PQRS TUV45,WX@. L.. " 4!

(304)

(305) YZ7[@. brum

(306)

(307) 7! D. edodes Leccinum. sca-. :;!\])5!B. Efg7(S]P5

(308) ]P !7. ^_`5 abDEcdefg. 7 $]Puv@kYZ. YZ Mana ῌ Ins ῌhPOiῌCer Man j. Type ῎ Man w, 3X@. e7 !7klm@no

(309) . "¡7 ] ¢£)* Can-.

(310) pF45qr ". tharellus cibarius 7@ (. klm stkuvw,῍Type. S]P~

(311) ]P~@ $]P. ῌ Gal/Fuc w ; Type ῍ Gal/Fuc/Man x. !F

(312)

(313) :;!,. vw Type ῎ Man w n7. 54¤¥@. 45yzr" GFig. ῌ῏I Type ῌ. AGLῌ a 7A, ab. Gal / Fuc w Type ῍ Gal / Fuc /. }""b7 , TLCῌimmunostaining. Man xvw7{9YZ, :;!|7. B¦û§!YZB!7@. }~ Type ῎ Man. " , TMS ¨5@. w, ] @. ~YF GC ©Bª} ©u. G ῍ I Type ῌ Gal/Fuc w,. §, AGLῌ a Man : Gal : Fuc G :. DEfg7(S]P

(314) ]P!Y. : I 5,«} Man : Gal : Fuc G :. F

(315)

(316) ] . : I}"445,AGLῌ a 7~. ῍ H. marmoreus. @¬B_`! AGLῌ b GGala ῌ. P.. Fig. ῌ῏ Three Types of Mycoglycolipids in Fungi.. NII-Electronic Library Service.

(317) Shiga University. 4U·¸y¹º»:¼no. Galb ῌ Mana ῌ Ins ῌPOῌCer

(318) AGLῌ AGLῌ b AGLῌ a Fuca ῌ Galb ῌ Mana ῌ Ins ῌ POῌCer TLC . AGLῌ a AGLῌ b !"#$ % &&'()*+,

(319) -. - /0123456789:); <$=>?@ABCD E FGH IJKFG LM NFG B OPO - ῍. ῌ. Q RS HT U / Foods Food Ingredients J. Jpn., ῎ῌ῕, ῌ . Q R. Jennemann, B. L. Bauer, H. Dertalanffy, R. Geyer, R. M. Gschwind, T. Selmer, H. Wiegandt Eur. J. Biochem., ῎ῑ῕, ῌ . Q R. Jennemann, R. Geyer, R. Sandhoff, R. M. Gschwind, S. B. Levery, HῌJ. Gröne, H. Wiegandt Eur. J. Biochem., ῎ῒ῔, ῌ . Q J. D. Weete Fungal Lipid Biochemistry, Chapter . Sphingolipids, pp. ῌ , Plenumpress, New York and London. Q B. Weiss, R. L. Stiller Biochemistry, ῍῍, ῌ . Q !"#V $%W XJYZ ῎ῑ,. ῌ Q. Q G. Kawai, Y. Ikeda Biochim. Biophys. Acta, ΐῑῐ, ῌ . Q R. L. Lester, R. C. Dickson Advances in Lipid Research, Vol. R. M. Bell, Y. A. Hannum, A. H. Merrill Jr., eds. pp. ῌ , Academic Press, San Diego. Q M. Ohnishi, S. Kawase, Y. Kondo, Y. Fujino, S. Ito J. Jpn. Oil Chem. Soc., ῐῑ, ῌ . Q RS [\& ]'^( HT U / _` JabJ@) c*I J ῑ῏, ῌ Q.

(320). Q RS [\& +,de HT f @g ]h- ./i U / _` JabJ@) c*IJ ῑῑ, ῌ Q Q M. Sugita, T. Mizunoma, K. Aoki, J. T. Dulaney, F. Inagaki, M. Suzuki, A. Suzuki, S. Ichikawa, K. Kushida, S. Ohta, A. Kurimoto Biochi. Biophys. Acta, ῍῏ῌ῎, ῌ . Q J. C. Dittmer, R. L. Lester J. Lipid Res., ῑ, ῌ . Q C. S. Hanes, F. A. Isherwood Nature, ῍ῒῐ, ῌ . Q I. Ciucanu, F. Kerek Carbohydr. Res., ῍῏῍, ῌ . Q S. Itonori, M. Takahashi, T. Kitamura, K. Aoki, J. T. Dulaney, M. Sugita J. Lipid Res., ῐῑ, ῌ . Q R. C. Gaver, C. C. Sweeley J. Am. Oil Chem. Soc., ῐ῎, ῌ . Q H T E j 0 k l m U / nopJFG ῐῌ, ῌ Q Q H T 1 2 q r ] s q t U / _` JabJ@) c*IJ ῐ῔, ῌ Q Q T. CῌY. Hsieh, K. Kaul, R. A. Laine, R. L. Lester Biochemistry, ῍ΐ, ῌ . Q u3v4 w(xyzx&{|;}~ {y; 56 78 09 H pp. ῌ qQ Q U / :; 7< ./i ,= >,5 ? @AB CpJ ῐ῍ ῌ Q Q RS DEF GH ?@ I 3J U / _` JabJ @) c*IJ ῑῒ, ῌ Q Q RS 9ῌK +,de DEF ?4 L@ 3MT N0 I 3J U / _` JabJ @) c*IJ ῑῒ, ῌ Q Q O@ +,de PQ 3MT ]h- RS U / R SCpJYTY ¡¢£ J ¤¥) ¦§ pp. Q Q Q T 3ÿ:©;ª«¬FG® ¯4UVWXYU+U -°

(321) 4ZX[±78²\³ ´]µ¶Q. NII-Electronic Library Service.

(322)