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Engineering a new pathway for N-acetylglucosamine production: Coupling a catabolic enzyme, glucosamine-6-phosphate deaminase, with a biosynthetic enzyme, glucosamine-6-phosphate N-acetyltransferase

Paper ID Volume ID Publish Year Pages File Format Full-Text
18528 42725 2006 7 PDF Available
Title
Engineering a new pathway for N-acetylglucosamine production: Coupling a catabolic enzyme, glucosamine-6-phosphate deaminase, with a biosynthetic enzyme, glucosamine-6-phosphate N-acetyltransferase
Abstract

A metabolic pathway for high level production of N-acetylglucosamine has been engineered in Escherichia coli by overexpressing E. coli glucosamine synthase (GlmS) and Saccharomyces cerevisiae glucosamine-6-phosphate acetyltransferase (GNA1). GlmS catalyzes the synthesis of glucosamine-6-phosphate from fructose-6-phosphate and glutamine. GNA1 converts glucosamine-6-phosphate into N-acetylglucosamine, which is dephosphorylated and secreted into the growth medium. In the present work, E. coli glucosamine-6-phosphate deaminase (NagB) was evaluated as an alternative to GlmS for the production of glucosamine and N-acetylglucosamine. NagB is a catabolic enzyme that converts glucosamine-6-phosphate to fructose-6-phosphate and ammonia. The reverse biosynthetic reaction forming glucosamine-6-phosphate is kinetically unfavorable. In a glmS deletion strain requiring glucosamine supplement to survive and grow, NagB overexpression resulted in the synthesis of glucosamine-6-phosphate. This supported cell growth, but little or no glucosamine accumulated in the medium. Overexpression of both NagB and GNA1 resulted in production of N-acetylglucosamine at levels comparable to strains overexpressing both GlmS and GNA1. This indicates that the overexpression of GNA1 played a critical role in determining the direction and efficiency of the reaction catalyzed by NagB. These data demonstrate that a catabolic enzyme can be utilized in a biosynthetic pathway by coupling with an efficient downstream reaction.

Keywords
Glucosamine; N-Acetylglucosamine; Escherichia coli; Glucosamine synthase; Glucosamine deaminase; Glucosamine-6-phosphate N-Acetyltransferase; Metabolic engineering
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Engineering a new pathway for N-acetylglucosamine production: Coupling a catabolic enzyme, glucosamine-6-phosphate deaminase, with a biosynthetic enzyme, glucosamine-6-phosphate N-acetyltransferase
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 39, Issue 4, 2 August 2006, Pages 828–834
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us