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Biosynthesis of nonnutritive monosaccharide d-allulose by metabolically engineered Escherichia coli from nutritive disaccharide sucrose
Biotechnology and Bioengineering ( IF 3.5 ) Pub Date : 2024-09-11 , DOI: 10.1002/bit.28842
Ling-Jie Zheng 1, 2 , Wei-Xiang Chen 1 , Shang-He Zheng 1 , Irfan Ullah 3 , Hui-Dong Zheng 1, 2 , Li-Hai Fan 1, 2 , Qiang Guo 1
Affiliation  

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.

中文翻译:


从营养二糖蔗糖中代谢工程大肠杆菌生物合成非营养性单糖 d-阿洛酮糖



蔗糖是食品和饮料中常用的营养甜味剂,因为它自然界含量丰富且生产成本低。然而,过量摄入蔗糖会增加代谢紊乱的风险,包括糖尿病和肥胖症。因此,对开发几乎没有卡路里的非营养性甜味剂的需求不断增长。d-阿洛酮糖是一种超低热量、稀有的六碳单糖,具有高甜度,使其成为蔗糖的理想替代品。在这项研究中,我们开发了一种细胞工厂,用于使用大肠杆菌 JM109 (DE3) 作为底盘宿主从蔗糖中生产 d-阿洛酮糖。基因 cscAcscBcscKalsEa6PP 共表达用于构建合成途径。然后,引入 ptsG-F 并敲除 ptsGfruAptsIptsH 以重编程糖转运途径,导致底物利用率的提高。接下来,Embden-Meyerhof-Parnas 的碳通量和磷酸戊糖途径受 pfkAzwf 失活的调控,d-阿洛酮糖滴度和产量分别提高了 154.2% 和 161.1%。最后,在 5 L 发酵罐中进行放大发酵。d-阿洛酮糖滴度达到 11.15 g/L,蔗糖收率为 0.208 g/g。
更新日期:2024-09-11
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