Metabolic engineering of Escherichia coli for industrial production of glucosamine and N-acetylglucosamine

被引：137

作者：

Deng, MD ^{[1
]}

Severson, DK ^{[1
]}

Grund, AD ^{[1
]}

Wassink, SL ^{[1
]}

Burlingame, RP ^{[1
]}

Berry, A ^{[1
]}

Running, JA ^{[1
]}

Kunesh, CA ^{[1
]}

Song, L ^{[1
]}

Jerrell, TA ^{[1
]}

Rosson, RA ^{[1
]}

机构：

[1] BioTech Resources, Manitowoc, WI 54220 USA

来源：

METABOLIC ENGINEERING | 2005年 / 7卷 / 03期

关键词：

glucosamine; N-acetylglucosamine; Esherichia coli; glmS gene; GNA1; gene; metabolic engineering; fermentation; lactose induction;

D O I：

10.1016/j.ymben.2005.02.001

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：

Glucosamine and N-acetylglucosamine are currently produced by extraction and acid hydrolysis of chitin from shellfish waste. Production could be limited by the amount of raw material available and the product potentially carries the risk of shellfish protein contamination. Escherichia coli was modified by metabolic engineering to develop a fermentation process. Over-expression of glucosamine synthase (GlmS) and inactivation of catabolic genes increased glucosamine production by 15 fold, reaching 60mgl(-1). Since GlmS is strongly inhibited by glucosamine-6-P, Gluts variants were generated via error-prone PCR and screened. Overexpression of an improved enzyme led to a glucosamine titer of 17 g l(-1). Rapid degradation of glucosamine and inhibitory effects of glucosamine and its degradation products on host cells limited further improvement. An alternative fermentation product, N-acetylglucosamine, is stable, non-inhibitory to the host and readily hydrolyzed to glucosamine under acidic conditions. Therefore, the glucosamine pathway was extended to N-acetylglucosamine by over-expressing a heterologous glucosamine-6-P N-acetyltransferase. Using a simple and low-cost fermentation process developed for this strain, over 110 gl(-1) of N-acetylglucosamine was produced. (c) 2005 Elsevier Inc. All rights reserved.

引用

页码：201 / 214

页数：14

共 39 条

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