De novo biosynthesis of 3-hydroxy-3-methylbutyrate as anti-catabolic supplement by metabolically engineered Escherichia coli

IF 6.8 1区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Sally J. Huang , Martin J. Lai , Arvin Y. Chen , Ethan I. Lan
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Abstract

3-Hydroxy-3-methylbutyrate (HMB) is a five-carbon branch-chain hydroxy acid currently used as a dietary supplement to treat sarcopenia and exercise training. However, its current production relies on conventional chemical processes which require toxic substances and are generally non-sustainable. While bio-based syntheses of HMB have been developed, they are dependent on biotransformation of its direct precursors which are generally costly. Therefore, in this work, we developed a synthetic de novo HMB biosynthetic pathway that enables HMB production from renewable resources. This novel HMB biosynthesis employs heterologous enzymes from mevalonate pathway and myxobacterial iso-fatty acid pathway for converting acetyl-CoA to HMB-CoA. Subsequently, HMB-CoA is hydrolyzed by a thioesterase to yield HMB. Upon expression of this pathway, our initial Escherichia coli strain produced 660 mg/L of HMB from glucose in 48 hours. Through optimization of coenzyme A removal from HMB-CoA and genetic operon structure, our final strain achieved HMB production titer of 17.7 g/L in glucose minimal media using a bench-top bioreactor. This engineered strain was further demonstrated to produce HMB from other renewable carbon sources such as xylose, glycerol, and acetate. The results from this work provided a flexible and environmentally benign method for producing HMB.

Abstract Image

3-羟基-3-甲基丁酸作为抗分解代谢补充剂的新生物合成。
3-羟甲基丁酸(HMB)是一种五碳支链羟基酸,目前被用作治疗肌肉疏松症和运动训练的膳食补充剂。然而,目前其生产依赖于传统的化学工艺,这些工艺需要使用有毒物质,而且通常是不可持续的。虽然已经开发出基于生物的 HMB 合成方法,但这些方法依赖于其直接前体的生物转化,而生物转化通常成本高昂。因此,在这项工作中,我们开发了一种新的合成 HMB 生物合成途径,能够利用可再生资源生产 HMB。这种新型的 HMB 生物合成利用了来自甲羟戊酸途径和肌杆菌异脂肪酸途径的异源酶,将乙酰-CoA 转化为 HMB-CoA。随后,HMB-CoA 被硫酯酶水解,生成 HMB。在表达这一途径后,我们最初的大肠杆菌菌株在 48 小时内从葡萄糖中产生了 660 毫克/升的 HMB。通过优化从 HMB-CoA 中去除辅酶 A 和基因操作子结构,我们的最终菌株在使用台式生物反应器的葡萄糖最小培养基中实现了 17.7 克/升的 HMB 产量。该工程菌株还进一步证明可利用木糖、甘油和醋酸等其他可再生碳源生产 HMB。这项工作的成果为生产 HMB 提供了一种灵活且对环境无害的方法。
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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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