文献类型：期刊文献

英文题名：Combinatorial metabolic engineering of Pseudomonas putida KT2440 for efficient mineralization of 1,2,3-trichloropropane

作者：Gong, Ting[1];Xu, Xiaoqing[1];Che, You[1];Liu, Ruihua[2];Gao, Weixia[1];Zhao, Fengjie[1];Yu, Huilei[3];Liang, Jingnan[4];Xu, Ping[5,6];Song, Cunjiang[1];Yang, Chao[1]

机构：[1]Nankai Univ, Minist Educ, Key Lab Mol Microbiol & Technol, Tianjin 300071, Peoples R China;[2]Nankai Univ, State Key Lab Med Chem Biol, Tianjin 300071, Peoples R China;[3]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[4]Chinese Acad Sci, Inst Microbiol, Core Facil Equipment, Beijing 100101, Peoples R China;[5]Shanghai Jiao Tong Univ, State Key Lab Microbial Metab, Shanghai 200240, Peoples R China;[6]Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai 200240, Peoples R China

年份：2017

卷号：7

外文期刊名：SCIENTIFIC REPORTS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000406764200096)】；

基金：This work was supported by the National Natural Science Foundation of China (No. 31570035), the National Key Technology Support Program of China (No. 2015BAD16B04), Open Fund of State Key Laboratory of Microbial Technology, Shandong University (No. M2016-03), and Open Fund of State Key Laboratory of Microbial Metabolism, Shanghai Jiao Tong University (No. MMLKF16-02).

语种：英文

摘要：An industrial waste, 1,2,3-trichloropropane (TCP), is toxic and extremely recalcitrant to biodegradation. To date, no natural TCP degraders able to mineralize TCP aerobically have been isolated. In this work, we engineered a biosafety Pseudomonas putida strain KT2440 for aerobic mineralization of TCP by implantation of a synthetic biodegradation pathway into the chromosome and further improved TCP mineralization using combinatorial engineering strategies. Initially, a synthetic pathway composed of haloalkane dehalogenase, haloalcohol dehalogenase and epoxide hydrolase was functionally assembled for the conversion of TCP into glycerol in P. putida KT2440. Then, the growth lag-phase of using glycerol as a growth precursor was eliminated by deleting the glpR gene, significantly enhancing the flux of carbon through the pathway. Subsequently, we improved the oxygen sequestering capacity of this strain through the heterologous expression of Vitreoscilla hemoglobin, which makes this strain able to mineralize TCP under oxygen-limited conditions. Lastly, we further improved intracellular energy charge (ATP/ADP ratio) and reducing power (NADPH/NADP(+) ratio) by deleting flagella-related genes in the genome of P. putida KT2440. The resulting strain (named KTU-TGVF) could efficiently utilize TCP as the sole source of carbon for growth. Degradation studies in a bioreactor highlight the value of this engineered strain for TCP bioremediation.