DETECTION O F FUCOSE FROM THE
POLYSACCHARIDES O F SOME JAPANESE LEGUMES
Takashi WAKITA(Laboratory of Agricultural Products Technology)
T h e presence of fucose a s one of the constituent sugars of hemicellulose
B1of broad bean seeds was found by K A W A M U R A and NARASAKI"-~'
.Further, t h e presence of fucose in t h e hemicellulose B1 of defatted soybean meal was confirmed by KAWAMURA and NARASAKI"). These findings opened a question whether fucose is a common constituent of legume hemicelluloses or a special component of relatively restricted species of legumes. T h e n t h e present experiment was undertaken t o clarify t h e distribution of fucose i n some represen. tative Japanese legumes. Kidney bean, pea, sword bean, and peanut were examined and i t was found t h a t all the hemicelluloses of these legumes contained fucose as a constiuent sugar.
1. Fractionation of Legume Polysacchsrides by Successive Extraction 1.1
Following four legumes were used as t h e sample:
1. Kidney bean, Phaseolus vulgarzs , "ingen-mame" In Japanese.
2. Pea, Pzsum satzvunz, "endo".
3 . Sword bean, Canavalza gladzata, "natamame".
4.P e a n u t , Arachzs h ypogaea , "r akkasei"
T h e y were a l l purchased on t h e market and i n excellent conditions to have complete germinating abilities.
2.Fractzonatzon o f P~lysaccharzdes
Seeds were dehulled, pulverized, and then defatted by extraction with ethyl e t h e r . T h e defatted seed powders were air dried and then subjected to fractionation of polysacharides according t o a scheme outlined in Fig 1.
1.3. Yzelds and Physzcal APPearances ofFractzonated Polysaccharzdes
Table I shows the yields of ~olysaccharides separated from each seeds. All t h e legumes
contained large amounts of polysaccharides soluble i n 5 % NaOH
Regardless of t h e species, all t h e hot-H20-soluble polysaccharides were brown and sticky, t h e 0.5 % (NH4)2C204-~01uble ones were white powders, the 0 2% N a O H ~ s o l u b l e ones were brown solids, and the 5 % NaOH-soluble ones were white powders.
1.4. Detectzon o f Component Sugars o f the Fvactzonated Polysaccharzdes by Paper Chromatography.
Tech Bull Fac Agr Kagawa Univ
Defatted seeds (208)
Extd with 200 mlH,O
609 4 h r s , 4 times
-IConed to v J10,
"i'Exth 200 m l 0 5% (NH,),C,O,, below 40"
, 170". 4 h r s , , 4 tzmes Add T C ~ to 2 5% S o l
P p t sol IC,.~ to ./lo
"'7Extd -,th 200 m1 o 2% N-H.
IAdd 3 vols EtOH P P ~
Et,O P P ~ wlth AcOH
Hot-H,O -sol Et,O
24 h r s . plysacchar ~ d e Add 3 vols o 5% (NH,),C,O, sol EtOH polysaeeharxde Made pH 6 8 wlth AcOH Coned to I Add 3 vols 0 2% NaCH sol polysaeeharxde
IEtOH P o t Et,O
1D r y 5% NaOH -sol
Fig 1 Fractionaion of polysacchaxides polysaccharide
Table I . Yields of polysaccharides
tube by heating in boiling water for 4 hrs. The hydrolyzate was neutralized with BaCOs up to pH 6.8 and the formed precipitate was removed by centrifugation and filtration through Toyo No. 5 C filter paper. The neutralized clear hydrolyzate was concentrated to dryness in vacuo below 4O0C and the residue was taken up in 1 ml 50
%aqueous ethanol to be examined by paper chromatography. Two-dimensional paper chromatography was carried out with phenol water (4 : 1) and n-butanol-pyridine- water (3 : 1 : 1.5) as the solvent systems and 0.3 %
P-anisidhe-HCI in water-saturated n-butanol as the spraying reagent. The results are shown in Tables 11-V.
Kidney bean Pea Sword bean Peanut
Polysacchaxides (g from 20 g of defatted seeds)
Hot-HzO-sol 3 2 1 6 3 2 2 1 5 % NaOH-sol 11 6 10 8 12 4 10 2 0 5% (NH4)eCzOs-sol 2 4 1 .O 3 0 2 8 0 2% NaOH-sol 1 9 0 4 0 8 0 8
Table I1 Component sugars of the kidney bean polysaccharides
Table I11 Component sugars of the pea polysaccharides
P o l y s a c c h a r i d e s ~
Rhamnose Fucose X y lose Ar abinose Galactose Glucose
+ +4- Hot-Hz0 0 5% (NH4)2C20a 0 2% NaOH 5 % NaOH Galacturonic Galactose
+- - F If-
+ +-1 Hot-Hz0 0 5% (NH4)2C204 0 2% NaOH 5 % NaOH
Table V Component sugars of the peanut polysaccharides
Table IV Component sugars of the sword bean polysaccharides
Rhamnose Fucose Xylose Arabinose Galactose Glucose
acid soluble in
1Glucose / ~ a l a c t u r o n i c
soluble in acid
+- - - - 0 2% NaOH
+5 % NaOH
-All the polysaccharides contained comparatively large amounts of glucose, arabinose, and galactose. Fucose was found only in the polysaccharides of kidney bean.
2 .Purification of the
%NaOH-soluble Polysaccharides by the Treatment with Taka-
Diastase and Detection of the Component Sugars of the Purified Legume Hemicelluloses
KAWAMURA et a1.C' ') reported that the crude hemicellulose B of broad bean seeds contained
only glucose and arabinose as revealed by paper chromatoglaphy. However, when t h e hemicellulose was purified by t h e treatment with Taka-diastase, glucose, galactose, xylose, and fucose were detected a s component sugars by paper chromatography T h i s indicates
- - -
+Hot-Hz0 0 5% (NH4)2C204 0 2% NaOH 5 % NaOH -- - - - - - -
+ +i- f +
++21 + + f F
118 Tech Bull Fac Agr Kagawa Univ.
t h a t t h e crude hemicellulose B of broad bean seeds consists largely of starch and araban
and these two polysaccbarides are removed by the treatment with Taka-diastase. Thus the authors purified t h e 5
%NaOH.soluble polysaccharides by the procedure of KAWAMURA
et al.C7) and the purified hemicelluloses were examined for their component sugaIs by paper
chromatography. T h e obtained results are given in Table VI. All the hemicelluloses contained rhamnose and fucose a s in t h e case of soybean hemicellulose BICO). These results
suggest that fucose is distributed widely in legume seed hemicelluloses as one of t h e common component sugars
Table VI Component sugars of the purified hemicelluloses
acid Acknowledgment Kidney bean Pea Sword bean Peanut -
-T h e authors wish to thank prof. Sin'itiro K A W A M U R A , Laboratory of Biological Chem- istr y , in the University, for his kind guidance. A part of expenditure was defrayed by t h e research fund donated by t h e Ministiry of Education to T . NARASAKI (Hemicelluloses of some Japanese legumes, 1960)
(1) KAWAMURA, S , NARASBKI, T
.Bull A g r . gaku, 9,889 (1960).
Chem Soc J a p a n , 22, 436 (1958). 15) NARASAKI, T Kagawa Datgaku Nogakubu
(2) KAWAMURA, S , NARASAKI, T : N t p p o n N o - Gakuzyutu Hokoku, 12, 109 (1960).
gei Kagaku Katshz, 33, 817 (1959). (6) KAWAMURA, S , NARASAKI , T
(3) NARASAKI, T. : Kagawa Dazgaku Nogakubu Chem , 25, 527 (1961)
Gakuzyutu Hokoku, 1 1 , 265 (1 959) (7) KAWAMURA, S., e t al : Kagawa Daigaku No-
(4) NARASAKI, T , KAWAMURA, S : Bunsekt K a - gakubu Gakuzyntu Hokoku, 8 , 201 (1957)
(Received October 27, 1965)