Plate assay for endo-β-N-acetylglucosaminidase activity using a chromogenic substrate synthesized by transglycosylation with Arthrobacter endo-β-N-acetylglucosaminidase

被引：4

作者：

Fujita, K

Asada, Y

Yamamoto, K

Takegawa, K ^{[1
]}

机构：

[1] Kagawa Univ, Fac Agr, Dept Life Sci, Miki, Kagawa 7610795, Japan

[2] Kyoto Univ, Grad Sch Agr, Div Appl Life Sci, Kyoto 6068502, Japan

来源：

JOURNAL OF BIOSCIENCE AND BIOENGINEERING | 2000年 / 90卷 / 04期

关键词：

endo-beta-N-acetylglucosaminidase; colorimetric detection; transglycosylation; X-Glc;

D O I：

10.1263/jbb.90.462

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

The transglycosylation activity of Arthrobacter endo-beta -N-acetylglucosaminidase (Endo-A) was used for the enzymatic synthesis of a novel oligosaccharide, Man(6)GlcNAc-5-bromo-4-chloro-3-indolyl-beta -glucoside (Man(6)GlcNAc-Glc-beta -X). Various endo-beta -N-acetylglucosaminidases hydrolyzed this oligosaccharide, producing Man(6)GlcNAc and Glc-beta -X. The E. coli strains coexpressing Endo-A and beta -glucosidase formed blue colonies in the presence of Man(6)GlcNAc-Glc-beta -X. Therefore, endo-beta -N-acetylglucosaminidase activity could be directly detected by the plate assay. This simple plate assay is useful for screening microorganisms for endo-beta -N-acetylglucosaminidase activity.

引用

收藏

页码：462 / 464

页数：3

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