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Title of Journal: Biotechnol Lett

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Abbravation: Biotechnology Letters

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Springer Netherlands

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ISSN

1573-6776

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NahK/GlmU fusion enzyme characterization and one

Authors: Yafei Zhai Min Liang Junqiang Fang Xiaoyue Wang Wanyi Guan Xianwei Liu Peng Wang Fengshan Wang
Publish Date: 2012/03/29
Volume: 34, Issue: 7, Pages: 1321-1326
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Abstract

The availability of uridine 5′diphosphate Nacetylglucosamine UDPGlcNAc is a prerequisite for the GlcNActransferasecatalyzed glycosylation reaction UDPGlcNAc has already been synthesized using an Nacetylhexosamine 1kinase NahK and a GlcNAc1P uridyltransferase truncated GlmU and here a fusion enzyme was constructed with truncated GlmU and NahK After determination of the optimum catalytic condition pH 80 at 40 °C the fusion enzyme was used to synthesize UDPGlcNAc in a single step with a yield of 88  from GlcNAc ATP and UTP Furthermore a simplified purification method was demonstrated using separation by gel filtration after byproduct digestion with alkaline phosphatase An overall yield of 77  and a purity of over 90  were achieved

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