Metabolic engineering of Halomonas bluephagenesis for production of five carbon molecular chemicals derived from L-lysine.

Autor: Yang F; School of Life Sciences, Tsinghua University, Beijing, 100084, China., Wang H; School of Life Sciences, Tsinghua University, Beijing, 100084, China., Zhao C; School of Life Sciences, Tsinghua University, Beijing, 100084, China., Zhang L; School of Life Sciences, Tsinghua University, Beijing, 100084, China., Liu X; PhaBuilder Biotech Co. Ltd., Shunyi District, Zhaoquan Ying, Beijing, 101309, China., Park H; School of Life Sciences, Tsinghua University, Beijing, 100084, China., Yuan Y; School of Life Sciences, Tsinghua University, Beijing, 100084, China., Ye JW; School of Life Sciences, Tsinghua University, Beijing, 100084, China., Wu Q; School of Life Sciences, Tsinghua University, Beijing, 100084, China., Chen GQ; School of Life Sciences, Tsinghua University, Beijing, 100084, China; Center for Synthetic and Systems Biology, Tsinghua University, Beijing, 100084, China; Tsinghua-Peking Center for Life Sciences, Beijing, China; MOE Key Lab of Industrial Biocatalysis, Dept Chemical Engineering, Tsinghua University, Beijing, 100084, China. Electronic address: chengq@mail.tsinghua.edu.cn.
Jazyk: angličtina
Zdroj: Metabolic engineering [Metab Eng] 2024 Jan; Vol. 81, pp. 227-237. Date of Electronic Publication: 2023 Dec 10.
DOI: 10.1016/j.ymben.2023.12.001
Abstrakt: 5-Aminovaleric acid (5-AVA), 5-hydroxyvalerate (5HV), copolymer P(3HB-co-5HV) of 3-hydroxybutyrate (3HB) and 5HV were produced from L-lysine as a substrate by recombinant Halomonas bluephagenesis constructed based on codon optimization, deletions of competitive pathway and L-lysine export protein, and three copies of davBA genes encoding L-lysine monooxygenase (DavB) and 5-aminovaleramide amidohydrolase (DavA) inserted into its genome to form H. bluephagenesis YF117ΔgabT 1+2 , which produced 16.4 g L -1 and 67.4 g L -1 5-AVA in flask cultures and in 7 L bioreactor, respectively. It was able to de novo synthesize 5-AVA from glucose by L-lysine-overproducing H. bluephagenesis TD226. Corn steep liquor was used instead of yeast extract for cost reduction during the 5-AVA production. Using promoter engineering based on P porin mutant library for downstream genes, H. bluephagenesis YF117 harboring pSEVA341-P porin 42 -yqhD EC produced 6 g L -1 5HV in shake flask growth, while H. bluephagenesis YF117 harboring pSEVA341-P porin 42 -yqhD EC -P porin 278 -phaC RE -abfT synthesized 42 wt% P(3HB-co-4.8 mol% 5HV) under the same condition. Thus, H. bluephagenesis was successfully engineered to produce 5-AVA and 5HV in supernatant and intracellular P(3HB-co-5HV) utilizing L-lysine as the substrate.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2023 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: