改造大肠杆菌,从甘油中生物合成 D-异戊糖。

IF 4.1 2区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Journal of biotechnology Pub Date : 2024-08-23 DOI:10.1016/j.jbiotec.2024.08.012
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引用次数: 0

摘要

D- 阿洛糖是一种天然单糖,在自然界中含量很少。由于它在消化道中的吸收率低,生长能量为零,因此被认为是一种有价值的低热量甜味剂。最近,生产 D-阿洛糖的大部分工作都集中在体外酶催化上。然而,微生物发酵正在成为一种有前途的替代方法,它具有在单个生物反应器内将酶制剂制造和产品合成相结合的优势。本文提出了一种利用代谢工程大肠杆菌从甘油中高效生物合成 D-异戊糖的新方法。FbaA、Fbp、AlsE 和 A6PP 被用来构建 D-异戊糖合成途径。随后,破坏了 PfkA、PfkB 和 Pgi,以阻止中间体 6-磷酸果糖(F6P)进入恩伯登-迈耶霍夫-帕尔纳斯(EMP)和磷酸戊糖(PP)途径。此外,GalE 和 FryA 被灭活,以减少细胞对 D-阿洛糖的消耗。最后,采用了喂料批次发酵工艺来优化细胞工厂的性能。结果,D-阿洛糖的滴度达到 7.02g/L,最高产量为 0.287g/g。
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Engineering Escherichia coli for D-allulose biosynthesis from glycerol

D-allulose, a naturally occurring monosaccharide, is present in small quantities in nature. It is considered a valuable low-calorie sweetener due to its low absorption in the digestive tract and zero energy for growth. Most of the recent efforts to produce D-allulose have focused on in vitro enzyme catalysis. However, microbial fermentation is emerging as a promising alternative that offers the advantage of combining enzyme manufacturing and product synthesis within a single bioreactor. Here, a novel approach was proposed for the efficient biosynthesis of D-allulose from glycerol using metabolically engineered Escherichia coli. FbaA, Fbp, AlsE, and A6PP were used to construct the D-allulose synthesis pathway. Subsequently, PfkA, PfkB, and Pgi were disrupted to block the entry of the intermediate fructose-6-phosphate (F6P) into the Embden−Meyerhof−Parnas (EMP) and pentose phosphate (PP) pathways. Additionally, GalE and FryA were inactivated to reduce D-allulose consumption by the cells. Finally, a fed-batch fermentation process was implemented to optimize the performance of the cell factory. As a result, the titer of D-allulose reached 7.02 g/L with a maximum yield of 0.287 g/g.

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来源期刊
Journal of biotechnology
Journal of biotechnology 工程技术-生物工程与应用微生物
CiteScore
8.90
自引率
2.40%
发文量
190
审稿时长
45 days
期刊介绍: The Journal of Biotechnology has an open access mirror journal, the Journal of Biotechnology: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. The Journal provides a medium for the rapid publication of both full-length articles and short communications on novel and innovative aspects of biotechnology. The Journal will accept papers ranging from genetic or molecular biological positions to those covering biochemical, chemical or bioprocess engineering aspects as well as computer application of new software concepts, provided that in each case the material is directly relevant to biotechnological systems. Papers presenting information of a multidisciplinary nature that would not be suitable for publication in a journal devoted to a single discipline, are particularly welcome.
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