Screening of Glucosamine-6-phosphate N-acetyltransferase for N-acetylglucosamine Production and Enzyme Kinetics Analysis

doi:10.3969/j.issn.1673-1689.2019.12.011

Home > Archive>Volume 38, Issue 12, 2019 >73-84. DOI:10.3969/j.issn.1673-1689.2019.12.011

Screening of Glucosamine-6-phosphate N-acetyltransferase for N-acetylglucosamine Production and Enzyme Kinetics Analysis
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                        10.3969/j.issn.1673-1689.2019.12.011
                    
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N-acetylglucosamine（GlcNAc），derivative of glucosamine（glucosamine，GlcN），is widely used in the field of health food and pharmaceutical. Glucosamine-6-phosphate N-acetyltransferase（GNA） is one of the crucial enzymes in the GlcNAc synthesis pathway. On the basis of recombinant GlcNAc-producing Bacillus subtilis，we overexpressed glucosamine-6-phosphate N-acetyltransferase，originates from Caenorhabditis elegans（Cegna1），in the engineered B. subtilis，which enhanced GlcNAc titer to 7.31 g/L. Compared with the engineered B. subtilis with overexpression of glucosamine-6-phosphate N-acetyltransferase derived from Saccharomyces cerevisiae，the GlcNAc yield of the recombinant B. subtilis with Cegna1 increased by 24.51%. By fed-batch fermentation in 3 L bioreactor，GlcNAc titer further increased to 24.23 g/L，which is 24.69% higher than the reference strain. Further investigation results showed that the recombinant Cegna1 has higher affinity and catalytic efficiency for GlcN-6P and Ac-CoA. Finally，it is found that the substrate GlcN-6P has obvious inhibition on the the recombinant Cegna1. Identified substrate inhibition of Cegna1 provides future directions for improvement of GlcNAc production by directed evolution of Cegna1 for alleviating substrate inhibition.

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XU Xiaofang, LIU Yanfeng, LI Jianghua, LIU Long, DU Guocheng, CHEN Jian. Screening of Glucosamine-6-phosphate N-acetyltransferase for N-acetylglucosamine Production and Enzyme Kinetics Analysis[J]. Journal of Food Science and Biotechnology,2019,38(12):73-84.

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