研究报告

CRISPRi干扰中心代谢基因转录对苏氨酸合成的影响

  • 刘旭峰 ,
  • 王宁 ,
  • 郝亚男 ,
  • 李英滋 ,
  • 范晓光 ,
  • 谢希贤
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  • 1(代谢控制发酵技术国家地方联合工程实验室(天津科技大学),天津,300457)
    2(天津市微生物代谢与发酵过程控制技术工程中心,天津,300457)
硕士研究生(谢希贤教授为通讯作者,E-mail:xixianxie@tust.edu.cn)。

收稿日期: 2018-11-29

  修回日期: 2019-01-17

  网络出版日期: 2019-05-14

基金资助

国家自然科学基金青年基金(31700037);工业微生物优良菌种选育与发酵技术公共服务平台项目(17PTGCCX00190)

Threonine synthesis under interfered transcriptions of genes involved in central metabolic pathway by CRISPRiLIU

  • Xufeng ,
  • WANG Ning ,
  • HAO Yanan ,
  • LI Yingzi ,
  • FAN Xiaoguang ,
  • XIE Xixian
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  • 1(National and Local United Engineering Lab of Metabolic Control Fermentation Technology (Tianjin University of Science and Technology), Tianjin 300457, China)
    2(Tianjin Engineering Research Center of Microbial Metabolism and FermentationProcess Control, Tianjin 300457, China)

Received date: 2018-11-29

  Revised date: 2019-01-17

  Online published: 2019-05-14

摘要

苏氨酸是重要的饲料氨基酸,需求量持续增加,提高苏氨酸发酵产率和糖酸转化率,降低生产成本已成为一个重要课题。该实验以苏氨酸工程菌Escherichia coli THRD为出发菌,利用CRISPRi(clustered regularly interspaced short palindromic repeats interference)技术研究中心代谢9个基因转录水平的改变对苏氨酸合成的影响。发酵结果显示,干扰zwf、pfkAgltA基因的转录水平提高了苏氨酸的合成效率,对应菌株苏氨酸产量分别为60.3、64.6和65.8 g/L,与出发菌(50.9 g/L)相比,分别提高了18.5%、26.9%和29.3%。糖酸转化率分别为40%、38%和39%,与出发菌(34%)相比,分别提高了17.7%、11.8%和14.7%。结果表明,通过CRISPRi干扰中心代谢基因的转录水平,可以调节合成代谢网络,使更多碳源流向苏氨酸,提高苏氨酸的合成效率。同时,该研究也为其他生物制品工程菌的构建提供了参考。

本文引用格式

刘旭峰 , 王宁 , 郝亚男 , 李英滋 , 范晓光 , 谢希贤 . CRISPRi干扰中心代谢基因转录对苏氨酸合成的影响[J]. 食品与发酵工业, 2019 , 45(8) : 1 -7 . DOI: 10.13995/j.cnki.11-1802/ts.019489

Abstract

Threonine is an important feed amino acid with increasing demand. Improving threonine productivity and glucose conversion rate and reducing the production costs have become an important subject. In this study, Escherichia coli THRD, a threonine-producing strain, was used to study the effects of different transcription levels of nine genes involved in central metabolic pathway on threonine synthesis by using clustered regularly interspaced short palindromic repeats interference (CRISPRi). The results showed that the transcriptional interference of genes zwf, pfkA, and gltA improved the synthesis efficiency of threonine. The threonine titers of corresponding strains were 60.3, 64.6, and 65.8 g/L, respectively, which were increased by 18.5%, 26.9%, and 29.3%, respectively, compared with the original strain (50.9 g/L). The glucose conversion rates were 40%, 38%, and 39%, respectively, which were 17.7%, 11.8%, and 14.7% higher than that of the original strain (34%). The results showed that by modifying transcriptional levels of genes that involved in central metabolism by CRISPRi could regulate the cellular metabolic network and redirect more carbon flux to threonine, resulted improved synthesis efficiency of threonine. Overall, this study provides a reference for constructing other bioengineered bacteria.

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