Detection of fucose from the polysaccharides of some Japanese legumes-香川大学学術情報リポジトリ

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DETECTION O F FUCOSE FROM THE

POLYSACCHARIDES O F SOME JAPANESE LEGUMES

Teiiti

NARASAKI

and

Takashi WAKITA

(Laboratory of Agricultural Products Technology)

T h e presence of fucose a s one of the constituent sugars of hemicellulose

B1

of 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.

Experimental

1. Fractionation of Legume Polysacchsrides by Successive Extraction 1.1

.

Samples

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.

1

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.

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Tech Bull Fac Agr Kagawa Univ

Defatted seeds (208)

Extd with 200 mlH,O

609 4 h r s , 4 times

7

Sol

I

-I

Coned to v J10,

"i'

Exth 200 m l 0 5% (NH,),C,O,, below 40"

, 1

70". 4 h r s , , 4 tzmes Add T C ~ to 2 5% S o l

r

P p t sol IC,.~ to ./lo

"'7

Extd -,th 200 m1 o 2% N-H.

I

Add 3 vols EtOH P P ~

EtOH

Et,O P P ~ wlth AcOH

Dry EtOH

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 _Add₃_vols 0 2% NaCH sol polysaeeharxde

I

EtOH P o t Et,O

1

D 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.

Legumes

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

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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 ~

1

Rhamnose Fucose X y lose Ar abinose Galactose Glucose

soluble in

Galacturonic acid

Polysaccharides

1

Rhamnose. Fucose

1

Xylose

1

Arabinose

soluble in

++

+

+ +

4- Hot-Hz0 0 5% (NH4)2C20a 0 2% NaOH 5 % NaOH Galacturonic Galactose

1

Glucose

1

acid

+

++

+

- - F If-

++

+++

+ +

-1 Hot-Hz0 0 5% (NH4)2C204 0 2% NaOH 5 % NaOH

Table V Component sugars of the peanut polysaccharides

+++

- If-

++

+ +

Table IV Component sugars of the sword bean polysaccharides

Polysaccharides Galacturonic

Rhamnose Fucose Xylose Arabinose Galactose Glucose

acid soluble in

PolysaccharidesI Rhamnose

I

Fucose

1

Xylose

1

Arabinose

1

Galactose

1

Glucose / ~ a l a c t u r o n i c

soluble in acid

+++

+

+++

+

- - - - 0 2% NaOH

++

+++

-5

+ +

+

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

5 %

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

- - -

++

+

-1

+++

+

-

+

Hot-Hz0 0 5% (NH4)2C204 0 2% NaOH 5 % NaOH -- - - - - - -

-

+

I

+

++

+ +

i- f +

++

21 + + f F

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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

Galactur onic

Legumes

)

Rbamnose

/

Fucose

1

Xylose

1

Arabinose

1

Galactose

1

Glucose

1

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)

.

Re ferenees

+

++

+

f

+

I

(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

.

Agr

.

Bzol

(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)