| 重组大肠杆菌1,2,4-丁三醇合成途径的平衡优化 |
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| 引用本文： | 郭超,冯奥,陆信曜,宗红,诸葛斌.重组大肠杆菌1,2,4-丁三醇合成途径的平衡优化[J].化工进展,2022,41(12):6531-6539. |
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| 作者姓名： | 郭超 冯奥 陆信曜 宗红 诸葛斌 |
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| 作者单位： | 江南大学工业生物技术教育部重点实验室，生物工程学院，工业微生物研究中心，江苏 无锡 214122 |
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| 摘 要： | 1,2,4-丁三醇（1,2,4-butantriol，BT）属于非天然化学品，是军工材料1,2,4-丁三醇三硝酸酯的前体。在重组大肠杆菌中，木糖经脱氢、脱水、脱羧和还原合成BT。但途径各反应不平衡使得中间代谢物积累限制菌体生长和产物合成。本研究首先通过CRISPR/Cas9敲除基因yjhG和yqhD构建无本底表达宿主菌，随后利用不同启动子组合调节BT合成途径中基因xdh、yjhG和yqhD的表达。结果发现，以P inv 表达醇脱氢酶基因yqhD使BT产量达到14.4g/L；以P tac 表达脱氢酶基因xdh和P rrnH P1表达脱水酶基因yjhG的组合方式，BT产量达到15.6g/L，比对照菌株KXW3009分别增加5.9%和14.7%。本研究通过对中间代谢物木糖酸合成和代谢的组合优化，促进了BT的合成，为后续研究提供了借鉴。
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| 关 键 词： | 1 2 4-丁三醇 大肠杆菌 启动子 途径平衡 |
| 收稿时间： | 2022-03-23 |
Balanced optimization of the 1,2,4-butanetriol synthesis pathway in recombinant Escherichia coli |
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| GUO Chao,FENG Ao,LU Xinyao,ZONG Hong,ZHUGE Bin.Balanced optimization of the 1,2,4-butanetriol synthesis pathway in recombinant Escherichia coli[J].Chemical Industry and Engineering Progress,2022,41(12):6531-6539. |
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| Authors: | GUO Chao FENG Ao LU Xinyao ZONG Hong ZHUGE Bin |
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| Affiliation: | Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Laboratory of Industrial Microorganisms, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China |
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| Abstract: | 1,2,4-Butantriol (BT) is a non-natural chemical product, which is the precursor of 1,2,4-butantriol trinitrate of military materials. In recombinant E. coli, BT was synthesized from xylose by dehydrogenation, dehydration, decarboxylation and reduction. However, the unbalanced reactions of the pathways lead to the accumulation of intermediate metabolites, which limits bacterial growth and product synthesis. In this study, CRISPR/Cas9 was used to knock out genes yjhG and yqhD to construct a backgroundless expression host bacteria, and then different promoter combinations were used to regulate the expression of genes xdh, yjhG and yqhD in the BT synthesis pathway. It was found that expression of the alcohol dehydrogenase gene yqhD with P inv resulted in BT yield of 14.4g/L and the combination of dehydrogenase gene xdh expressed by P tac and dehydrase gene yjhG expressed by P rrnH P1 made BT yield reach 15.6g/L, which was 5.9% and 14.7% higher than that of the control strain KXW3009, respectively. This study facilitated the synthesis of BT through the regulation of upstream and downstream gene expression of intermediate metabolites, which provides a reference for subsequent studies. |
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| Keywords: | 1 2 4-butanetriol Escherichia coli promoter balance pathway |
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