Engineering carbon source division of labor for efficient α-carotene production in Corynebacterium glutamicum

IF 6.8 1区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Kai Li , Cheng Li , Chen-Guang Liu , Xin-Qing Zhao , Ruiwen Ou , Charles A. Swofford , Feng-Wu Bai , Gregory Stephanopoulos , Anthony J. Sinskey
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Abstract

Effective utilization of glucose, xylose, and acetate, common carbon sources in lignocellulose hydrolysate, can boost biomanufacturing economics. However, carbon leaks into biomass biosynthesis pathways instead of the intended target product remain to be optimized. This study aimed to enhance α-carotene production by optimizing glucose, xylose, and acetate utilization in a high-efficiency Corynebacterium glutamicum cell factory. Heterologous xylose pathway expression in C. glutamicum resulted in strain m4, exhibiting a two-fold increase in α-carotene production from xylose compared to glucose. Xylose utilization was found to boost the biosynthesis of pyruvate and acetyl-CoA, essential precursors for carotenoid biosynthesis. Additionally, metabolic engineering including pck, pyc, ppc, and aceE deletion, completely disrupted the metabolic connection between glycolysis and the TCA cycle, further enhancing α-carotene production. This strategic intervention directed glucose and xylose primarily towards target chemical production, while acetate supplied essential metabolites for cell growth recovery. The engineered strain C. glutamicum m8 achieved 30 mg/g α-carotene, 67% higher than strain m4. In fed-batch fermentation, strain m8 produced 1802 mg/L of α-carotene, marking the highest titer reported to date in microbial fermentation. Moreover, it exhibited excellent performance in authentic lignocellulosic hydrolysate, producing 216 mg/L α-carotene, 1.45 times higher than the initial strain (m4). These labor-division strategies significantly contribute to the development of clean processes for producing various valuable chemicals from lignocellulosic resources.

在谷氨酸棒杆菌中进行碳源分工,以高效生产 α-胡萝卜素。
有效利用木质纤维素水解物中的常见碳源--葡萄糖、木糖和醋酸,可以提高生物制造的经济效益。然而,碳泄漏到生物质生物合成途径而非预期目标产品的情况仍有待优化。本研究旨在通过优化高效谷氨酸棒杆菌细胞工厂中葡萄糖、木糖和醋酸盐的利用,提高α-胡萝卜素的产量。谷氨酸棒状杆菌中木糖途径的异源表达导致菌株 m4 的木糖α-胡萝卜素产量比葡萄糖高出三倍。研究发现,木糖的利用促进了丙酮酸和乙酰-CoA(类胡萝卜素生物合成的重要前体)的生物合成。此外,包括 pck、pyc、ppc 和 aceE 缺失在内的代谢工程完全破坏了糖酵解和 TCA 循环之间的代谢联系,进一步提高了 α 胡萝卜素的产量。这种战略性干预将葡萄糖和木糖主要用于目标化学品的生产,而乙酸则为细胞恢复生长提供必需的代谢物。工程菌株 C. glutamicum m8 的 α-胡萝卜素产量为 30 mg/g,比菌株 m4 高出 67%。在饲料批量发酵中,菌株 m8 产生了 1,802 毫克/升的α-胡萝卜素,这是迄今为止微生物发酵中报道的最高滴度。此外,它在真正的木质纤维素水解物中表现优异,产生了 216 mg/L α-胡萝卜素,是初始菌株(m4)的 1.75 倍。这些分工策略极大地促进了利用木质纤维素资源生产各种有价值化学品的清洁工艺的发展。
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来源期刊
Metabolic engineering
Metabolic engineering 工程技术-生物工程与应用微生物
CiteScore
15.60
自引率
6.00%
发文量
140
审稿时长
44 days
期刊介绍: Metabolic Engineering (MBE) is a journal that focuses on publishing original research papers on the directed modulation of metabolic pathways for metabolite overproduction or the enhancement of cellular properties. It welcomes papers that describe the engineering of native pathways and the synthesis of heterologous pathways to convert microorganisms into microbial cell factories. The journal covers experimental, computational, and modeling approaches for understanding metabolic pathways and manipulating them through genetic, media, or environmental means. Effective exploration of metabolic pathways necessitates the use of molecular biology and biochemistry methods, as well as engineering techniques for modeling and data analysis. MBE serves as a platform for interdisciplinary research in fields such as biochemistry, molecular biology, applied microbiology, cellular physiology, cellular nutrition in health and disease, and biochemical engineering. The journal publishes various types of papers, including original research papers and review papers. It is indexed and abstracted in databases such as Scopus, Embase, EMBiology, Current Contents - Life Sciences and Clinical Medicine, Science Citation Index, PubMed/Medline, CAS and Biotechnology Citation Index.
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