Multivariate modular metabolic engineering and medium optimization for vitamin B12 production by Escherichia coli

IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Synthetic and Systems Biotechnology Pub Date : 2024-04-04 DOI:10.1016/j.synbio.2024.03.017
Feitao Chen , Huan Fang , Jianghua Zhao , Pingtao Jiang , Huina Dong , Ying Zhao , Huiying Wang , Tongcun Zhang , Dawei Zhang
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Abstract

Vitamin B12 is a complex compound synthesized by microorganisms. The industrial production of vitamin B12 relies on specific microbial fermentation processes. E. coli has been utilized as a host for the de novo biosynthesis of vitamin B12, incorporating approximately 30 heterologous genes. However, a metabolic imbalance in the intricate pathway significantly limits vitamin B12 production. In this study, we employed multivariate modular metabolic engineering to enhance vitamin B12 production in E. coli by manipulating two modules comprising a total of 10 genes within the vitamin B12 biosynthetic pathway. These two modules were integrated into the chromosome of a chassis cell, regulated by T7, J23119, and J23106 promoters to achieve combinatorial pathway optimization. The highest vitamin B12 titer was attained by engineering the two modules controlled by J23119 and T7 promoters. The inclusion of yeast powder to the fermentation medium increased the vitamin B12 titer to 1.52 mg/L. This enhancement was attributed to the effect of yeast powder on elevating the oxygen transfer rate and augmenting the strain's isopropyl-β-d-1-thiogalactopyranoside (IPTG) tolerance. Ultimately, vitamin B12 titer of 2.89 mg/L was achieved through scaled-up fermentation in a 5-liter fermenter. The strategies reported herein will expedite the development of industry-scale vitamin B12 production utilizing E. coli.

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大肠杆菌生产维生素 B12 的多元模块化代谢工程和培养基优化
维生素 B12 是一种由微生物合成的复杂化合物。维生素 B12 的工业生产依赖于特定的微生物发酵过程。大肠杆菌被用作维生素 B12 从头生物合成的宿主,其中包含约 30 个异源基因。然而,复杂途径中的代谢失衡极大地限制了维生素 B12 的生产。在这项研究中,我们采用了多元模块化代谢工程技术,通过操纵维生素 B12 生物合成途径中由总共 10 个基因组成的两个模块来提高大肠杆菌的维生素 B12 产量。这两个模块被整合到底盘细胞的染色体中,由 T7、J23119 和 J23106 启动子调控,以实现组合途径优化。通过设计由 J23119 和 T7 启动子控制的两个模块,获得了最高的维生素 B12 滴度。在发酵培养基中加入酵母粉后,维生素 B12 滴度增至 1.52 毫克/升。这种提高归因于酵母粉提高了氧转移率并增强了菌株对异丙基-β-d-1-硫代吡喃半乳糖苷(IPTG)的耐受性。最终,通过在 5 升发酵罐中进行放大发酵,维生素 B12 的滴度达到了 2.89 毫克/升。本文报告的策略将加快利用大肠杆菌进行工业规模维生素 B12 生产的发展。
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来源期刊
Synthetic and Systems Biotechnology
Synthetic and Systems Biotechnology BIOTECHNOLOGY & APPLIED MICROBIOLOGY-
CiteScore
6.90
自引率
12.50%
发文量
90
审稿时长
67 days
期刊介绍: Synthetic and Systems Biotechnology aims to promote the communication of original research in synthetic and systems biology, with strong emphasis on applications towards biotechnology. This journal is a quarterly peer-reviewed journal led by Editor-in-Chief Lixin Zhang. The journal publishes high-quality research; focusing on integrative approaches to enable the understanding and design of biological systems, and research to develop the application of systems and synthetic biology to natural systems. This journal will publish Articles, Short notes, Methods, Mini Reviews, Commentary and Conference reviews.
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