Effect of Pathway Configurations on the Synthesis of 2'-Fucosyllactose in Escherichia coli

被引：0
|
作者：
Liang S. [1 ]
Zhang D. [1 ]
Yang S. [1 ]
He Z. [1 ]
Liu D. [1 ]
Jiang Z. [1 ]
机构：
[1] Key Laboratory of Food Bioengineering, China National Light Industry, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing
来源：
Shipin Kexue/Food Science | 2022年 / 43卷 / 24期
关键词：
2'-fucosyllactose; Biosynthesis; CRISPR/Cas9; Escherichia coli; Pathway configurations;
D O I：
10.7506/spkx1002-6630-20220602-016
中图分类号：
学科分类号：
摘要：
The de novo synthesis pathway of 2'-fucosyllactose (2'-FL) was established in Escherichia coli BL21 Star (DE3)in this study. The β-galactosidase gene lacZ M15 and the uridine diphosphate (UDP)-glucose lipid carrier transferase gene wcaJ were knocked out using the CRISPR/Cas9 gene editing system. The effects of three different pathway configurations, viz., operon, pseudo-operon, and monocistronic on the synthesis of 2'-FL were explored. The results showed that the concentration of 2'-FL produced in shake flasks was 0.34 g/L after overexpression of the de novo synthesis pathway related genes in E. coli BL21 Star (DE3). The concentration of 2'-FL was increased to 1.26 g/L by deleting the lacZ M15 and wcaJ genes. The highest concentration of 2'-FL of 1.92 g/L was observed in strain BS-7 when regulated by the operon expression. Fed-batch fermentation of strain BS-7 accumulated 14.04 g/L 2'-FL with a productivity of 0.59 g/(L•h) and a lactose conversion rate of 63%, respectively. This study suggested that lower gene expression levels not only increased 2'-FL production, but also could improve the conversion efficiency of substrate in engineered E. coli. © 2022, China Food Publishing Company. All right reserved.
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页码：110 / 116
页数：6
相关论文
共 27 条
  • [1] BALLARD O, MORROW A L., Human milk composition: nutrients and bioactive factors, Pediatric Clinics, 60, 1, pp. 49-74, (2013)
  • [2] BYCH K, MIKS M H, JOHANSON T, Et al., Production of HMOs using microbial hosts: from cell engineering to large scale production, Current Opinion in Biotechnology, 56, pp. 130-137, (2019)
  • [3] LI W, WANG J, LIN Y, Et al., How far is it from infant formula to human milk? A look at the human milk oligosaccharides, Trends in Food Science & Technology, 118, pp. 374-387, (2021)
  • [4] FAIJES M, CASTEJON-VILATERSANA M, VAL-CID C, Et al., Enzymatic and cell factory approaches to the production of human milk oligosaccharides, Biotechnology Advances, 37, 5, pp. 667-697, (2019)
  • [5] ALBERMANN C, DISTLER J, PIEPERSBERG W., Preparative synthesis of GDP-β-L-fucose by recombinant enzymes from enterobacterial sources, Glycobiology, 10, 9, pp. 875-881, (2000)
  • [6] COYNE M J, REINAP B, LEE M M, Et al., Human symbionts use a host-like pathway for surface fucosylation[J], Science, 307, pp. 1778-1781, (2005)
  • [7] LEE W H, PATHANIBUL P, QUARTERMAN J, Et al., Whole cell biosynthesis of a functional oligosaccharide, 2'-fucosyllactose, using engineered Escherichia coli[J], Microbial Cell Factories, 11, 1, pp. 1-9, (2012)
  • [8] HUANG D, YANG K X, LIU J, Et al., Metabolic engineering of Escherichia coli for the production of 2'-fucosyllactose and 3-fucosyllactose through modular pathway enhancement, Metabolic Engineering, 41, pp. 23-38, (2017)
  • [9] CHIN Y W, SEO N, KIM J H, Et al., Metabolic engineering of Escherichia coli to produce 2'-fucosyllactose via salvage pathway of guanosine 5'-diphosphate (GDP)-l-fucose, Biotechnology and Bioengineering, 113, 11, pp. 2443-2452, (2016)
  • [10] LI W, ZHU Y, WAN L, Et al., Pathway optimization of 2'-fucosyllactose production in engineered Escherichia coli, Journal of Agricultural and Food Chemistry, 69, 5, pp. 1567-1577, (2021)
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