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	Metabolic engineering of Escherichia coli for biosynthesis of hyaluronic acid
	Yu, Huimin ; Stephanopoulos, Gregory
	2010-05-07 ; 2010-05-07
关键词	Hyaluronic acid Recombinant E. coli Artificial operon Rare codon Metabolic pathway Metabolic engineering CARBON FLUX DISTRIBUTIONS CORYNEBACTERIUM-GLUTAMICUM STREPTOCOCCUS-PYOGENES LYSINE OVERPRODUCTION MOLECULAR-CLONING BRANCH POINT SYNTHASES GENE IDENTIFICATION EQUISIMILIS Biotechnology & Applied Microbiology
中文摘要	Engineering of hyaluronic acid (HA) biosynthetic pathway in recombinant Escherichia coli as production host is reported in this work. A hyaluronic acid synthase (HAS) gene, sphasA, from Sreptococcus pyogenes with the start codon gtg to atg mutant, was expressed in recombinant E. coli with or without the genes ugd, galF and glmU, which are analogs of hasB, hasC and hasD from Streptococcus, respectively, encoding UDP-glucose 6-dehygrogenase, Glucose-1-P uridyltransferase, and N-acetyl glucosamine uridyltransferase enzymes in the HA biosynthetic pathway. The single, double and triple organized artificial operons of sphasA, ugd, galF and glmU were designed and constructed using the inducible plasmid backbone of pMBAD. Only the triple expression recombinant, Top10/pMBAD-spABC, generated a relatively high titer of HA (similar to 48 mg/l at 48 h), indicating that both of the enzymes encoded by ugd and galF are essential for HA biosynthesis. A new gene of ssehasA with identical protein sequence of seHAS from Streptococcus equisimilis, was artificially synthesized after substituting all of the rare codons in the natural sehasA. The HA titer at 24 h. ask culture increased to similar to 190 mg/l in sseAB and 160 mg/l in sseABC, respectively. Sorbitol could be used as another carbon source for HA accumulation, and the metabolic pathway for HA synthesis in a recombinant E. coli was presented. The concentration of Mg2+ cofactor of HA synthase was optimized and a cell growth inhibition phenomenon was observed during HA accumulation. Molecular weight (MW) measurements revealed that the mean MW of HA produced from the recombinant E. coli under different conditions ranges from similar to 3.5 x 10(5) to 1.9 x 10(6) Da, indicating that the recombinant E. coli can be used as a potential host candidate for industrial production of HA. (C) 2007 Elsevier Inc. All rights reserved.
语种	英语 ; 英语
出版者	ACADEMIC PRESS INC ELSEVIER SCIENCE ; SAN DIEGO ; 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA
内容类型	期刊论文
源URL	[http://hdl.handle.net/123456789/17407]
专题	清华大学
推荐引用方式 GB/T 7714	Yu, Huimin,Stephanopoulos, Gregory. Metabolic engineering of Escherichia coli for biosynthesis of hyaluronic acid[J],2010, 2010.
APA	Yu, Huimin,&Stephanopoulos, Gregory.(2010).Metabolic engineering of Escherichia coli for biosynthesis of hyaluronic acid..
MLA	Yu, Huimin,et al."Metabolic engineering of Escherichia coli for biosynthesis of hyaluronic acid".(2010).

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