Purification and characterization of alpha-L-fucosidase from Streptomyces species.

M Sano,K Hayakawa,I Kato

doi:10.1016/s0021-9258(18)45977-7

Abstract

Streptomyces sp. 142, isolated from a soil sample, produced alpha-fucosidase when cultured in the presence of L-fucose. The enzyme was purified 700-fold with an overall recovery of 17% from a cell-free extract by cation exchange chromatography and gel filtration chromatography. The apparent molecular weight of the purified enzyme was 40,000 by gel filtration chromatography. The enzyme had a pH optimum of 6.0 and was stable at pH 4.5-7.0. Substrate specificity studies with oligosaccharides labeled with 2-aminopyridine as the substrate showed that the enzyme specifically hydrolyzed terminal alpha 1-3 and alpha 1-4 fucosidic linkages in the oligosaccharides but did not hydrolyze alpha 1-2 or alpha 1-6 fucosidic linkages or a synthetic substrate, p-nitro-phenyl alpha-L-fucoside. The purified enzyme released L-fucose from a fucosylated glycoprotein, alpha 1-acid glycoprotein. Thus, the substrate specificities of the Streptomyces alpha-fucosidase resembled those of alpha-fucosidases I and III isolated from almond emulsin rather than those of other microbial alpha-fucosidases.

Highlights

Drolyzed terminal 011-3and a l - 4 fucosidic linkages in the oligosaccharides but did not hydrolyze a1-2 or a l - MATERIALS AND METHODS’
CI-L-FUCOrSe~s~idues are frequently found at thenonreducing termini of the carbohydrate part of glycoconjugates on mammalian cell surfaces and in gastric and submaxillary mucin, blood group substances, glycolipids, and serum glycoproteins
The metabolism of free L-fucose (Fuc)’ andthe glycoconjugatescontaining a-L-fucosyl residues are different with different diseases, including cancer [1,2,3,4,5]

Summary

Introduction

Drolyzed terminal 011-3and a l - 4 fucosidic linkages in the oligosaccharides but did not hydrolyze a1-2 or a l - MATERIALS AND METHODS’. The substrate specificity of the enzyme differs depending on its origin. The enzymes from mammalian tissues[7,8,9] and marine gastropods (10,ll) have extremely broad substrate specificities, but theenzymes from microorganisms [12,13,14,15] and almond emulsin [16] cleave only Fucd-2Gal. Two a-fucosidases with different substrate specificities have been isolated from Bacillus circulans M28 [17, 18].One hydrolyzes al-2, al-3, anda l - 4 fucosidic linkages, it does not act on a l - 6 fucosidic linkages.

Results

Conclusion