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公开(公告)号:US10731185B2
公开(公告)日:2020-08-04
申请号:US16086883
申请日:2017-03-21
发明人: Xianghao Wu , Mark Eiteman
摘要: Provided herein is a genetically engineered microbe which accumulates citramalate. In one embodiment, the microbe includes an exogenous polynucleotide encoding a citramalate synthase which catalyzes the condensation of acetyl CoA and pyruvic acid. Optionally, the microbe also includes a second exogenous polynucleotide encoding a citrate synthase which catalyzes the condensation of acetyl CoA and oxaloacetate, and the citrate synthase activity in the microbe is reduced compared to a control microbe. In one embodiment, the citrate synthase includes at least one amino acid substitution in the acetyl-CoA binding pocket, the mobile loop, the NADH binding site, and the oxaloacetate binding site, or a combination thereof. Also provided herein are methods for using the genetically engineered microbe, including a method for producing citramalate. The method can further include isolating the citramalate.
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公开(公告)号:US20190093135A1
公开(公告)日:2019-03-28
申请号:US16086883
申请日:2017-03-21
发明人: Xianghao Wu , Mark Eiteman
CPC分类号: C12P7/46 , C12N9/10 , C12N9/1025 , C12N9/1029 , C12N15/70 , C12Y203/01182 , C12Y203/03001
摘要: Provided herein is a genetically engineered microbe which accumulates citramalate. In one embodiment, the microbe includes an exogenous polynucleotide encoding a citramalate synthase which catalyzes the condensation of acetyl CoA and pyruvic acid. Optionally, the microbe also includes a second exogenous polynucleotide encoding a citrate synthase which catalyzes the condensation of acetyl CoA and oxaloacetate, and the citrate synthase activity in the microbe is reduced compared to a control microbe. In one embodiment, the citrate synthase includes at least one amino acid substitution in the acetyl-CoA binding pocket, the mobile loop, the NADH binding site, and the oxaloacetate binding site, or a combination thereof. Also provided herein are methods for using the genetically engineered microbe, including a method for producing citramalate. The method can further include isolating the citramalate.
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