Lu Jingxian, Li Zhimin, Ye Qin, Wu Hui
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
Sheng Wu Gong Cheng Xue Bao. 2019 Jan 25;35(1):59-69. doi: 10.13345/j.cjb.180107.
Poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)] belongs to the polyhydroxyalkanoates (PHA) family and possesses promising properties including biocompatibility and biodegradability. In this study, we directly synthesized P(3HB-co-LA) with glucose by introducing the β-ketothiolase and acetoacetyl-CoA reductase from Ralstonia eutropha, the engineered propionate CoA transferase from Clostridium propionicum and the engineered polyhydroxyalkanoate synthase from Pseudomonas fluorescens strain 2P24 into Escherichia coli. The polymer content was 83.9% (W/W), and the molar percentage of lactate reached 1.6%. On this basis, in order to accumulate lactate, we reduced the activity of respiratory chain by deleting the ubiX gene, which is involved in the synthesis of coenzyme Q8. Moreover, we removed the dld gene to avoid the conversion of lactate to pyruvate during the fermentation. With these manipulations, the molar percentage of lactate in the polymer was improved to 14.1%, with an 81.7% (W/W) of polymer content. The test results indicated that the strategy of reducing the activity of respiratory chain effectively increased the lactate units in the polymer, and it contributed a new approach to change the content of monomer components in the polymer.
聚(3-羟基丁酸酯-共-乳酸酯)[P(3HB-co-LA)]属于聚羟基脂肪酸酯(PHA)家族,具有包括生物相容性和生物降解性在内的良好性能。在本研究中,我们通过将来自真养产碱菌的β-酮硫解酶和乙酰乙酰辅酶A还原酶、来自丙酸梭菌的工程化丙酸辅酶A转移酶以及来自荧光假单胞菌2P24菌株的工程化聚羟基脂肪酸酯合酶导入大肠杆菌,直接利用葡萄糖合成了P(3HB-co-LA)。聚合物含量为83.9%(W/W),乳酸的摩尔百分比达到1.6%。在此基础上,为了积累乳酸,我们通过删除参与辅酶Q8合成的ubiX基因来降低呼吸链的活性。此外,我们去除了dld基因以避免发酵过程中乳酸转化为丙酮酸。通过这些操作,聚合物中乳酸的摩尔百分比提高到了14.1%,聚合物含量为81.7%(W/W)。测试结果表明,降低呼吸链活性的策略有效地增加了聚合物中的乳酸单元,为改变聚合物中单体组分的含量提供了一种新方法。