代谢工程大肠杆菌从营养性二糖蔗糖生物合成非营养性单糖 d-阿洛糖

IF 3.5 2区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Biotechnology and Bioengineering Pub Date : 2024-09-11 DOI:10.1002/bit.28842
Ling-Jie Zheng, Wei-Xiang Chen, Shang-He Zheng, Irfan Ullah, Hui-Dong Zheng, Li-Hai Fan, Qiang Guo
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引用次数: 0

摘要

蔗糖是食品和饮料中常用的营养甜味剂,因为它在自然界中含量丰富,生产成本低。然而,过量摄入蔗糖会增加患糖尿病和肥胖症等代谢性疾病的风险。d-Allulose 是一种超低卡路里的稀有六碳单糖,甜度高,是蔗糖的理想替代品。在这项研究中,我们以大肠杆菌 JM109 (DE3) 为底盘宿主,开发了从蔗糖生产 d-阿洛糖的细胞工厂。基因 cscA、cscB、cscK、alsE 和 a6PP 共同表达,构建了合成途径。然后,通过引入 ptsG-F 和敲除 ptsG、fruA、ptsI 和 ptsH 来重新规划糖运输途径,从而提高了底物利用率。接着,通过使 pfkA 和 zwf 失活来调节恩布登-梅耶霍夫-帕尔纳斯途径和磷酸戊糖途径的碳通量,使 d-阿洛糖滴度和产量分别提高了 154.2% 和 161.1%。最后,在 5 升发酵罐中进行了放大发酵。d-allulose 滴度达到 11.15 g/L,蔗糖产量为 0.208 g/g。
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Biosynthesis of nonnutritive monosaccharide d-allulose by metabolically engineered Escherichia coli from nutritive disaccharide sucrose

Sucrose is a commonly utilized nutritive sweetener in food and beverages due to its abundance in nature and low production costs. However, excessive intake of sucrose increases the risk of metabolic disorders, including diabetes and obesity. Therefore, there is a growing demand for the development of nonnutritive sweeteners with almost no calories. d-Allulose is an ultra-low-calorie, rare six-carbon monosaccharide with high sweetness, making it an ideal alternative to sucrose. In this study, we developed a cell factory for d-allulose production from sucrose using Escherichia coli JM109 (DE3) as a chassis host. The genes cscA, cscB, cscK, alsE, and a6PP were co-expressed for the construction of the synthesis pathway. Then, the introduction of ptsG-F and knockout of ptsG, fruA, ptsI, and ptsH to reprogram sugar transport pathways resulted in an improvement in substrate utilization. Next, the carbon fluxes of the Embden-Meyerhof-Parnas and the pentose phosphate pathways were regulated by the inactivation of pfkA and zwf, achieving an increase in d-allulose titer and yield of 154.2% and 161.1%, respectively. Finally, scaled-up fermentation was performed in a 5 L fermenter. The titer of d-allulose reached 11.15 g/L, with a yield of 0.208 g/g on sucrose.

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来源期刊
Biotechnology and Bioengineering
Biotechnology and Bioengineering 工程技术-生物工程与应用微生物
CiteScore
7.90
自引率
5.30%
发文量
280
审稿时长
2.1 months
期刊介绍: Biotechnology & Bioengineering publishes Perspectives, Articles, Reviews, Mini-Reviews, and Communications to the Editor that embrace all aspects of biotechnology. These include: -Enzyme systems and their applications, including enzyme reactors, purification, and applied aspects of protein engineering -Animal-cell biotechnology, including media development -Applied aspects of cellular physiology, metabolism, and energetics -Biocatalysis and applied enzymology, including enzyme reactors, protein engineering, and nanobiotechnology -Biothermodynamics -Biofuels, including biomass and renewable resource engineering -Biomaterials, including delivery systems and materials for tissue engineering -Bioprocess engineering, including kinetics and modeling of biological systems, transport phenomena in bioreactors, bioreactor design, monitoring, and control -Biosensors and instrumentation -Computational and systems biology, including bioinformatics and genomic/proteomic studies -Environmental biotechnology, including biofilms, algal systems, and bioremediation -Metabolic and cellular engineering -Plant-cell biotechnology -Spectroscopic and other analytical techniques for biotechnological applications -Synthetic biology -Tissue engineering, stem-cell bioengineering, regenerative medicine, gene therapy and delivery systems The editors will consider papers for publication based on novelty, their immediate or future impact on biotechnological processes, and their contribution to the advancement of biochemical engineering science. Submission of papers dealing with routine aspects of bioprocessing, description of established equipment, and routine applications of established methodologies (e.g., control strategies, modeling, experimental methods) is discouraged. Theoretical papers will be judged based on the novelty of the approach and their potential impact, or on their novel capability to predict and elucidate experimental observations.
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