文献类型：期刊文献

中文题名：丙酰辅酶A的体外酶法合成

英文题名：In vitro enzyme catalytic synthesis of propionyl-CoA

作者：聂雪彤[1];吴辉[1];李志敏[1];叶勤[1]

机构：[1]华东理工大学生物工程学院生物反应器国家重点实验室,上海200237

年份：2016

卷号：43

期号：6

起止页码：78

中文期刊名：北京化工大学学报（自然科学版）

外文期刊名：Journal of Beijing University of Chemical Technology(Natural Science Edition)

收录：CSTPCD;;Scopus;北大核心:【北大核心2014】；CSCD:【CSCD2015_2016】；

基金：国家"863"计划(2012AA022104)

语种：中文

中文关键词：大肠杆菌;丙酰-CoA;琥珀酸;高效液相色谱(HPLC)

外文关键词：Escherichia coli; propionyl-CoA; succinic acid; high performance liquid chromatography （HPLC）

摘要：丙酰辅酶A(丙酰-CoA)是一种重要的代谢中间物,可以用来生产丙烯酸、3-羟基丙酸、β-羟基戊酸和聚酮化合物。将天然的琥珀酰辅酶A合成酶、甲基丙二酰辅酶A变位酶、甲基丙二酰辅酶A脱羧酶在大肠杆菌中克隆并过表达,采用这3种纯化的酶体外催化合成丙酰-CoA,评价了琥珀酸生产丙酰-CoA代谢途径。琥珀酸可经琥珀酰辅酶A合成酶催化生成琥珀酰-CoA,琥珀酰-CoA再由甲基丙二酰辅酶A变位酶催化生成甲基丙二酰-CoA,最后甲基丙二酰-CoA经甲基丙二酰辅酶A脱羧酶催化生成丙酰-CoA。反应产物经高效液相色谱分析,检测到目标产物丙酰-CoA,结果证明了从琥珀酸体外合成丙酰-CoA的代谢途径的可行性。
Propionyl-CoA is an important metabolic intermediate for the production of acrylate, 3-hydroxypropi- onate, β-hydroxyvalerate and polyketides. The native succinyl-CoA synthetase, methylmalonyl-CoA mutase and methylmalonyl-CoA decarboxylase were cloned and overexpressed in Escherichia coll. These three enzymes were pu- rified and employed to produce propionyl-CoA in vitro. The metabolic pathway from succinic acid to propionyl-CoA was evaluated. Succinic acid was firstly converted into succinyl-CoA by succinyl-CoA synthetase; and the succinyl- CoA was sequentially converted into methylmalonyl-CoA by methylmalonyl-CoA mutase; finally, methylmalonyl- CoA was converted into propionyl-CoA by methylmalonyl-CoA decarboxylase. Propionyl-CoA in the reaction solu- tion was detected by high performance liquid chromatography （HPLC） , which demonstrate that the metabolic path- way from succinic acid to propionyl-CoA is feasible.