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AssigneesHenan Julong Biological Engineering Co., Ltd, Tianjin University of Science and Technology
InventorsXiaoguang Fan, Xiaodong Liu, Jing Li, Ning Chen, Bochao Liu, Shuai Liu, Chaochao Sun, Yongchao Liu, Jiajia Teng, Mengtao Zhang, Yuanqing Ji, Yuhang Zhou, Qingyang Xu
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The present invention belongs to the bioengineering field, and relates to a method for fermentation production of L-theanine by using an Escherichia coli genetically engineered bacterium. The engineered bacterium is obtained by serving a strain as an original strain, wherein the strain is obtained after performing a single copy of T7RNAP, a dual copy of gmas, xylR knockout, and sucCD knockout on an Escherichia coli W3110 genome, and by integrating genes xfp, pta, acs, gltA, and ppc, and knocking out ackA on the genome. The present invention has a high yield, and stable production performance; after 20-25 h, L-theanine has a titer of 75-80 g/L, and the yield is up to 52-55%. The fermentation broth is purified by membrane separation in combination with a cation-anion resin series technique. Moreover, the one-step crystallization yield is 72.3% and the L-theanine final product has a purity of 99%.
A genetically engineered bacterium producing L-theanine, wherein the genetically engineered bacterium is obtained by using a strain as an original strain, wherein the original strain is obtained after integrating a single copy of a RNA polymerase gene T7RNAP, a dual copy of a γ-glutamylmethylamide synthetase gene gmas, and a knockout of a xylose operon transcription factor gene xylR and a knockout of a succinyl-CoA synthetase gene sucCD on a genome of Escherichia coli W3110, and then by integrating an exogenous fructose 6-phosphate phosphoketolase gene xfp, an exogenous phosphoacetyl transferase gene pta, an exogenous acetyl-CoA synthetase gene acs, an exogenous citrate synthase gene gltA, and an exogenous phosphoenolpyruvate carboxylase gene ppc on the genome, and knocking out an acetokinase gene ackA.
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