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重组大肠杆菌共表达德氏乳杆菌丙二醇利用酶生产 3-羟基丙酸。

Production of 3-hydroxypropionic acid from 3-hydroxypropionaldehyde by recombinant Escherichia coli co-expressing Lactobacillus reuteri propanediol utilization enzymes.

机构信息

Department of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, P.O. Box 124, SE-221 00 Lund, Sweden.

Department of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, P.O. Box 124, SE-221 00 Lund, Sweden.

出版信息

Bioresour Technol. 2015 Mar;180:214-21. doi: 10.1016/j.biortech.2014.12.109. Epub 2015 Jan 7.

Abstract

3-Hydroxypropionic acid (3-HP) is an important platform chemical for the biobased chemical industry. Lactobacillus reuteri produces 3-HP from glycerol via 3-hydroxypropionaldehyde (3-HPA) through a CoA-dependent propanediol utilization (Pdu) pathway. This study was performed to verify and evaluate the pathway comprising propionaldehyde dehydrogenase (PduP), phosphotransacylase (PduL), and propionate kinase (PduW) for formation of 3-HP from 3-HPA. The pathway was confirmed using recombinant Escherichia coli co-expressing PduP, PduL and PduW of L. reuteri DSM 20016 and mutants lacking expression of either enzyme. Growing and resting cells of the recombinant strain produced 3-HP with a yield of 0.3mol/mol and 1mol/mol, respectively, from 3-HPA. 3-HP was the sole product with resting cells, while growing cells produced 1,3-propanediol as co-product. 3-HP production from glycerol was achieved with a yield of 0.68mol/mol by feeding recombinant E. coli with 3-HPA produced by L. reuteri and recovered using bisulfite-functionalized resin.

摘要

3-羟基丙酸(3-HP)是生物基化学工业中一种重要的平台化学品。乳杆菌通过依赖 CoA 的丙二醇利用(Pdu)途径将甘油转化为 3-羟基丙醛(3-HPA),从而产生 3-HP。本研究旨在验证和评估包含丙醛脱氢酶(PduP)、磷酸转酰酶(PduL)和丙酸激酶(PduW)的途径,以从 3-HPA 形成 3-HP。该途径通过共表达 L. reuteri DSM 20016 的 PduP、PduL 和 PduW 的重组大肠杆菌和缺乏任何一种酶表达的突变体来确认。重组菌株的生长和静止细胞分别以 0.3mol/mol 和 1mol/mol 的产率从 3-HPA 产生 3-HP。静止细胞的产物为 3-HP,而生长细胞的产物为 1,3-丙二醇。通过向重组大肠杆菌中添加由 L. reuteri 产生并使用亚硫酸氢盐功能化树脂回收的 3-HPA,以 0.68mol/mol 的产率从甘油生产 3-HP。

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