Donor substrate promiscuity of bacterial β1-3-N-acetylglucosaminyltransferases and acceptor substrate flexibility of β1-4-galactosyltransferases

被引：48

作者：

Li, Yanhong ^{[1
]}

Xue, Mengyang ^{[1
,2
,3
,6
]}

Sheng, Xue ^{[1
,7
]}

Yu, Hai ^{[1
]}

Zeng, Jie ^{[1
,4
]}

Thon, Vireak ^{[1
,5
,8
]}

Chen, Yi ^{[1
]}

Muthana, Musleh M. ^{[1
,9
]}

Wang, Peng G. ^{[2
,3
,5
]}

Chen, Xi ^{[1
]}

机构：

[1] Univ Calif Davis, Dept Chem, One Shields Ave, Davis, CA 95616 USA

[2] Shandong Univ, Natl Glycoengn Res Ctr, Jinan 250100, Shandong, Peoples R China

[3] Shandong Univ, Shandong Prov Key Lab Carbohydrate Chem & Glycobi, Jinan 250100, Shandong, Peoples R China

[4] Henan Inst Sci & Technol, Sch Food Sci, Xinxiang 453003, Henan, Peoples R China

[5] Georgia State Univ, Dept Chem, Atlanta, GA 30303 USA

[6] Shandong Univ, Shandong Prov Key Lab Emergency & Crit Care Med, Jinan 250100, Shandong, Peoples R China

[7] Univ Calif San Diego, Dept Chem & Biochem, San Diego, CA 92093 USA

[8] US FDA, Lab Bacterial Polysaccharides, Bethesda, MD 20892 USA

[9] Childrens Natl Med Ctr, Washington, DC 20010 USA

来源：

BIOORGANIC & MEDICINAL CHEMISTRY | 2016年 / 24卷 / 08期

基金：

美国国家卫生研究院; 美国国家科学基金会;

关键词：

Carbohydrate; Glycosyltransferase; Donor substrate promiscuity; Acceptor substrate specificity; Enzymatic synthesis; One-pot multienzyme; HUMAN-MILK OLIGOSACCHARIDES; O-LINKED GLYCANS; CHEMICAL-CHARACTERIZATION; CHEMOENZYMATIC SYNTHESIS; NEISSERIA-MENINGITIDIS; HELICOBACTER-PYLORI; N-ACETYLGLUCOSAMINE; ESCHERICHIA-COLI; EXPRESSION; BETA-1,4-GALACTOSYLTRANSFERASE;

D O I：

10.1016/j.bmc.2016.02.043

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

beta 1-3-N-Acetylglucosaminyltransferases (beta 3GlcNAcTs) and beta 1-4-galactosyltransferases (beta 4GalTs) have been broadly used in enzymatic synthesis of N-acetyllactosamine (LacNAc)-containing oligosaccharides and glycoconjugates including poly-LacNAc, and lacto-N-neotetraose (LNnT) found in the milk of human and other mammals. In order to explore oligosaccharides and derivatives that can be synthesized by the combination of beta 3GlcNAcTs and beta 4GalTs, donor substrate specificity studies of two bacterial beta 3GlcNAcTs from Helicobacter pylori (Hp beta 3GlcNAcT) and Neisseria meningitidis (NmLgtA), respectively, using a library of 39 sugar nucleotides were carried out. The two beta 3GlcNAcTs have complementary donor substrate promiscuity and 13 different trisaccharides were produced. They were used to investigate the acceptor substrate specificities of three beta 4GalTs from Neisseria meningitidis (NmLgtB), Helicobacter pylori (Hp beta 4GalT), and bovine (B beta 4GalT), respectively. Ten of the 13 trisaccharides were shown to be tolerable acceptors for at least one of these beta 4GalTs. The application of NmLgtA in one-pot multienzyme (OPME) synthesis of two trisaccharides including GalNAc beta 1-3Gal beta 1-4Glc beta ProN(3) and Gal beta 1-3Gal beta 1-4Glc was demonstrated. The study provides important information for using these glycosyltransferases as powerful catalysts in enzymatic and chemoenzymatic syntheses of oligosaccharides and derivatives which can be useful probes and reagents. (C) 2016 Elsevier Ltd. All rights reserved.

引用

页码：1696 / 1705

页数：10

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