精品范文2005 N-乙酰氨基葡萄糖.pdfVIP

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精品范文2005 N-乙酰氨基葡萄糖.pdf

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ARTICLE IN PRESS Metabolic Engineering 7 (2005) 201–214 /locate/ymben Metabolic engineering of Escherichia coli for industrial production of glucosamine and N-acetylglucosamine Ming-De Deng ,David K. Severson ,Alan D. Grund ,Sarah L. Wassink Richard P. Burlingame1 ,Alan Berry2 ,Jeffrey A. Running ,Candice A. Kunesh Linsheng Song ,Thomas A. Jerrell ,Reinhardt A. Rosson Bio-Technical Resources, 1035 South 7th Street, Manitowoc, WI 54220, USA Received 17 September 2004; accepted 8 February 2005 Available online 23 March 2005 Abstract 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 60 mg l1. Since GlmS is strongly inhibited by glucosamine-6-P ,GlmS variants were generated via error-prone PCR and screened. Over- expression of an improved enzyme led to a glucosamine titer of 17 g l1. Rapid degradation of glucosamine and inhibitory effects of glucosamine and its degradation products on host cells limited further impr