NahK/GlmU fusion enzyme: characterization and one-step enzymatic synthesis of UDP-N-acetylglucosamine

作者：Zhai Yafei; Liang Min; Fang Junqiang; Wang Xiaoyue; Guan Wanyi; Liu Xian wei^*; Wang Peng; Wang Fengshan

来源：Biotechnology Letters, 2012, 34(7): 1321-1326.

DOI：10.1007/s10529-012-0910-y

• N-Acetylglucosamine-1-phosphate uridyltransferase
• N-Acetylhexosamine 1-kinase
• Alkaline phosphatase
• Uridine 5 '-diphosphate N-acetylglucosamine

The availability of uridine 5'-diphosphate N-acetylglucosamine (UDP-GlcNAc) is a prerequisite for the GlcNAc-transferase-catalyzed glycosylation reaction. UDP-GlcNAc has already been synthesized using an N-acetylhexosamine 1-kinase (NahK) and a GlcNAc-1-P uridyltransferase (truncated GlmU) and here, a fusion enzyme was constructed with truncated GlmU and NahK. After determination of the optimum catalytic condition (pH 8.0 at 40 A degrees C), the fusion enzyme was used to synthesize UDP-GlcNAc 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 by-product digestion with alkaline phosphatase. An overall yield of 77 % and a purity of over 90 % were achieved.

• 出版日期2012-7
• 单位山东大学