Chemical Industry and Engineering Progress ›› 2024, Vol. 43 ›› Issue (2): 1063-1068.DOI: 10.16085/j.issn.1000-6613.2023-0298

• Biochemical and pharmaceutical engineering • Previous Articles    

Genetic engineering modification to promote the synthesis of 1,2,4-butanetriol from cellulose hydrolysate of Escherichia coli

SHE Dan(), WANG Shuting, LU Xinyao, ZONG Hong, ZHUGE Bin()   

  1. Key Laboratory of Industrial Biotechnology, Ministry of Education, Laboratory of Industrial Microorganisms, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China
  • Received:2023-03-01 Revised:2023-08-15 Online:2024-03-07 Published:2024-02-25
  • Contact: ZHUGE Bin

基因工程改造促进大肠杆菌发酵纤维素水解液合成1,2,4-丁三醇

佘丹(), 王舒婷, 陆信曜, 宗红, 诸葛斌()   

  1. 江南大学生物工程学院,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
  • 通讯作者: 诸葛斌
  • 作者简介:佘丹(1998—),女,硕士研究生,研究方向为微生物代谢工程。E-mail:2744167136@qq.com

Abstract:

1,2,4-Butanetriol (BT) is one of the high value-added compounds widely used in various fields, and the production of BT by fermentation using cellulose hydrolysate requires a strain that can tolerate the inhibitors present in the hydrolysate. In the presence of furfural, a common inhibitor in hydrolysates, the fermentation of Escherichia coli is severely limited. In this study, four furfural tolerance genes, yghA, thyA, mdtJI, and pntAB were overexpressed in the BT-producingstrain. In the presence of 0.4g/L furfural, all modified strains showed different degrees of growth. Among them, the strain overexpressing pntAB had the best growth, which also contributed to the enzymatic activity or transcriptional level of the enzymes in the BT pathway, yielding 11.0g/L BT. When corncob cellulose hydrolysate was used as the substrate, the strains overexpressing yghA, thyA, and pntAB all significantly improved the growth and BT production. The strain carrying yghA performed the best, yielding 3.36g/L BT in shake flasks and 15.3g/L BT in 5L fermenters, with a conversion of 63.8%. This strategy provided a reference for BT production by fermentation of corncob cellulose hydrolysate.

Key words: Escherichia coli, corncob cellulose hydrolysate, furfural, tolerance, 1,2,4-butanetriol

摘要:

1,2,4-丁三醇(BT)是广泛应用于各领域的高附加值化合物之一,利用纤维素水解液发酵生产BT需要一株能够耐受其中抑制物的菌株。在水解液中常见抑制因子糠醛存在下,大肠杆菌的发酵严重受限。本研究首先在含有BT合成途径的大肠杆菌中过表达四种糠醛耐受相关的基因:yghAthyAmdtJIpntAB。在0.4g/L糠醛压力下,所有改造菌株的生长均有不同程度的提升,其中过表达pntAB基因的菌株生长最好,同时有助于BT合成酶的酶活力或转录水平的提高,BT产量达到11.0g/L。当以玉米芯纤维素水解液为底物时,过表达yghAthyApntAB均明显促进了大肠杆菌的生长与BT产量的提高,其中yghA基因表现最好,在摇瓶中获得了3.36g/L的BT,在5L发酵罐中BT产量达到15.3g/L,转化率为63.8%。该策略为玉米芯纤维素水解液发酵生产BT提供了借鉴。

关键词: 大肠杆菌, 玉米芯纤维素水解液, 糠醛, 耐受, 1,2,4-丁三醇

CLC Number: 

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