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工程化乳酸乳球菌中R-和S-1,2-丙二醇的生产。

Production of R- and S-1,2-propanediol in engineered Lactococcus lactis.

作者信息

Sato Rintaro, Ikeda Motoyuki, Tanaka Tomonari, Ohara Hitomi, Aso Yuji

机构信息

Department of Biobased Materials Science, Kyoto Institute of Technology, Kyoto, Japan.

JST-Mirai Program, Japan Science and Technology Agency, Saitama, Japan.

出版信息

AMB Express. 2021 Aug 16;11(1):117. doi: 10.1186/s13568-021-01276-8.

DOI:10.1186/s13568-021-01276-8
PMID:34398341
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8368392/
Abstract

1,2-propanediol (1,2-PDO) is a versatile chemical used in multiple manufacturing processes. To date, some engineered and non-engineered microbes, such as Escherichia coli, Lactobacillus buchneri, and Clostridium thermosaccharolyticum, have been used to produce 1,2-PDO. In this study, we demonstrated the production of R- and S-1,2-PDO using engineered Lactococcus lactis. The L- and D-lactic acid-producing L. lactis strains NZ9000 and AH1 were transformed with the plasmid pNZ8048-ppy harboring pct, pduP, and yahK genes for 1,2-PDO biosynthesis, resulting in L. lactis LL1 and LL2, respectively. These engineered L. lactis produced S- and R-1,2-PDO at concentrations of 0.69 and 0.50 g/L with 94.4 and 78.0% ee optical purities, respectively, from 1% glucose after 72 h of cultivation. Both 1% mannitol and 1% gluconate were added instead of glucose to the culture of L. lactis LL1 to supply NADH and NADPH to the 1,2-PDO production pathway, resulting in 75% enhancement of S-1,2-PDO production. Production of S-1,2-PDO from 5% mannitol and 5% gluconate was demonstrated using L. lactis LL1 with a pH-stat approach. This resulted in S-1,2-PDO production at a concentration of 1.88 g/L after 96 h of cultivation. To our knowledge, this is the first report on the production of R- and S-1,2-PDO using engineered lactic acid bacteria.

摘要

1,2 - 丙二醇(1,2 - PDO)是一种用于多种制造工艺的通用化学品。迄今为止,一些工程菌和非工程菌,如大肠杆菌、布氏乳杆菌和嗜热解糖梭菌,已被用于生产1,2 - PDO。在本研究中,我们展示了利用工程化乳酸乳球菌生产R - 和S - 1,2 - PDO。将携带用于1,2 - PDO生物合成的pct、pduP和yahK基因的质粒pNZ8048 - ppy分别转化到产L - 和D - 乳酸的乳酸乳球菌菌株NZ9000和AH1中,分别得到乳酸乳球菌LL1和LL2。这些工程化的乳酸乳球菌在培养72小时后,从1%的葡萄糖中分别以0.69和0.50 g/L的浓度产生S - 和R - 1,2 - PDO,对映体过量光学纯度分别为94.4%和78.0%。向乳酸乳球菌LL1的培养物中添加1%的甘露醇和1%的葡萄糖酸盐代替葡萄糖,为1,2 - PDO生产途径提供NADH和NADPH,导致S - 1,2 - PDO产量提高75%。使用乳酸乳球菌LL1通过pH值控制法展示了从5%的甘露醇和5%的葡萄糖酸盐生产S - 1,2 - PDO。培养96小时后,这导致S -

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/799b/8368392/0309ab7bf6ff/13568_2021_1276_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/799b/8368392/f418d7429eeb/13568_2021_1276_Fig1_HTML.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/799b/8368392/d735a4e1ac70/13568_2021_1276_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/799b/8368392/2fe01163fd19/13568_2021_1276_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/799b/8368392/0309ab7bf6ff/13568_2021_1276_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/799b/8368392/f418d7429eeb/13568_2021_1276_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/799b/8368392/dcd9d0d9335b/13568_2021_1276_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/799b/8368392/d735a4e1ac70/13568_2021_1276_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/799b/8368392/2fe01163fd19/13568_2021_1276_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/799b/8368392/0309ab7bf6ff/13568_2021_1276_Fig5_HTML.jpg

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