食品与发酵工业 >

2021 , Vol. 47 >Issue 19: 29 - 34

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.026826

研究报告

大肠杆菌乙酸弱化对1,2,4-丁三醇合成的影响

  • 狄莹莹 ,
  • 王昕钰 ,
  • 冯奥 ,
  • 陆信曜 ,
  • 宗红 ,
  • 诸葛斌
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  • 1(糖化学与生物技术教育部重点实验室(江南大学),江苏 无锡,214122)
    2(江南大学,工业生物技术教育部重点实验室,工业微生物研究中心,江苏 无锡,214122)
硕士研究生(诸葛斌教授为通讯作者,E-mail:bzhuge@126.com)

收稿日期: 2021-01-26

  修回日期: 2021-02-26

  网络出版日期: 2021-11-04

基金资助

国家自然科学基金项目(21708016)

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Improved 1,2,4-butanetriol production by weaken acetate pathway in a recombinant Escherichia coli

  • DI Yingying ,
  • WANG Xinyu ,
  • FENG Ao ,
  • LU Xinyao ,
  • ZONG Hong ,
  • ZHUGE Bin
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  • 1(The Key Laboratory of Carbohydrate Chemistry and Biotechnology,Ministry of Education,Jiangnan University,Wuxi 214122,China)
    2(The Key Laboratory of Industrial Biotechnology,Ministry of Education,Jiangnan University,School of Biotechnology,Research Centre of Industrial Microbiology,Wuxi 214122,China)

Received date: 2021-01-26

  Revised date: 2021-02-26

  Online published: 2021-11-04

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摘要

以大肠杆菌(Escherichia coli)为宿主合成1,2,4-丁三醇,存在乙酸溢流,影响细胞生长和所需还原力。采用敲除E.coli丙酮酸氧化酶(PoxB)和磷酸转乙酰酶(Pta),弱化丙酮酸和乙酰辅酶A节点处的碳流引入乙酸的策略,考察细胞生长及1,2,4-丁三醇合成。结果显示:在E.coli中单敲pta或poxB及双敲对宿主生长都有影响,只有单敲poxB生物量提高了26.6%;将乙酸弱化菌株作为宿主细胞用于1,2,4-丁三醇合成,单敲pta或poxB基因均可以降低乙酸积累,提高1,2,4-丁三醇单位菌体产量,单敲poxB时1,2,4-丁三醇产量提高最大,达到13.1 g/L,提高了37.9%;双敲pta和poxB基因,乙酸进一步被弱化,菌体生长受到影响,但1,2,4-丁三醇单位菌体产量与对照相比提高最明显,提高了30%;5 L发酵罐分批补料发酵结果表明,单敲poxB菌株乙酸合成减少了34.4%,生物量提高了14.2%,1,2,4-丁三醇产量提高到16.1 g/L,木糖摩尔转化率为0.69 mol/mol。以上结果表明,适宜的乙酸途径弱化策略可以增强1,2,4-丁三醇合成能力,过度抑制乙酸途径反而会影响菌体生长。

关键词： pta; poxB; 乙酸; 1,2,4-丁三醇; 细胞生长

本文引用格式

狄莹莹 , 王昕钰 , 冯奥 , 陆信曜 , 宗红 , 诸葛斌 . 大肠杆菌乙酸弱化对1,2,4-丁三醇合成的影响[J]. 食品与发酵工业, 2021 , 47(19) : 29 -34 . DOI: 10.13995/j.cnki.11-1802/ts.026826

Abstract

The synthesis of 1,2,4-butanetriol (BT) in Escherichia coli suffers from acetate overflow, which affects cell growth and redox homeostasis. To reduce acetate production and to enhance cell growth and BT synthesis, the carbon flow at the nodes of pyruvate and acetyl coenzyme reduced by knocking out pyruvate oxidase (PoxB) and phosphotransacetylase (Pta). The results showed that deletion of pta and poxB alone, or both, had an effect on growth. The Acetate-weakened strain with pta or poxB knocked reduced the accumulation of acetate and increased the unit yield of BT. The strain knocked with pta and poxB had the weakened content of acetate and biomass, and the unit yield of BT increased by 30% compared to the control. The poxB knocked strain showed 34.4% reduction in acetate synthesis, 14.2% increase in biomass, 16.1 g/L production of BT, and a molar conversion of xylose of 0.69 mol/mol in 5 L fermenter. The results suggest that appropriate acetate pathway weakening strategies can enhance BT synthesis, while excessive inhibition of the acetate pathway can affect growth.

Key words： pta; poxB; acetate; 1,2,4-butanetriol; cell growth

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