Enzymatic production of α-d-galactose 1-phosphate by lactose phosphorolysis

被引：20

作者：

De Groeve, Manu R. M. ^{[1
]}

Depreitere, Veerle ^{[1
]}

Desmet, Tom ^{[1
]}

Soetaert, Wim ^{[1
]}

机构：

[1] Univ Ghent, Dept Biochem & Microbial Technol, Lab Ind Microbiol & Biocatalysis, B-9000 Ghent, Belgium

来源：

BIOTECHNOLOGY LETTERS | 2009年 / 31卷 / 12期

关键词：

Bioconversion; Galactose; 1-phosphate; Lactose phosphorylase; Phosphorolysis; Purification; Synthesis; CELLOBIOSE PHOSPHORYLASE; DIRECTED EVOLUTION; BIOTRANSFORMATION; PHOSPHATASE;

D O I：

10.1007/s10529-009-0087-1

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：

alpha-d-Galactose 1-phosphate (alpha Gal1P) is an important building block for the synthesis of nucleotide sugars that are substrates for glycosyltransferases. We have previously reported the creation of novel lactose phosphorylase enzymes that are useful for the synthesis of alpha Gal1P from the cheap and abundant lactose. Here we describe the application of such a lactose phosphorylase in a production system using permeabilized Escherichia coli cells. After purification of the product from the reaction mixture by anion-exchange chromatography and ethanol precipitation, 9.5 grams of highly pure alpha Gal1P were obtained from a 1 l reaction volume.

引用

页码：1873 / 1877

页数：5

共 17 条

[1] Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants:: the Keio collection [J].