Fang Yang , Huan Wang , Cuihuan Zhao , Lizhan Zhang , Xu Liu , Helen Park , Yiping Yuan , Jian-Wen Ye , Qiong Wu , Guo-Qiang Chen
{"title":"利用卤单胞菌的代谢工程生产源自 L-赖氨酸的五碳分子化学品","authors":"Fang Yang , Huan Wang , Cuihuan Zhao , Lizhan Zhang , Xu Liu , Helen Park , Yiping Yuan , Jian-Wen Ye , Qiong Wu , Guo-Qiang Chen","doi":"10.1016/j.ymben.2023.12.001","DOIUrl":null,"url":null,"abstract":"<div><p>5-Aminovaleric acid (5-AVA), 5-hydroxyvalerate (5HV), copolymer P(3HB-<em>co</em>-5HV) of <em>3</em>-hydroxybutyrate (3HB) and 5HV were produced from L-lysine as a substrate by recombinant <em>Halomonas bluephagenesis</em> constructed based on codon optimization, deletions of competitive pathway and L-lysine export protein, and three copies of <em>davBA</em> genes encoding L-lysine monooxygenase (DavB) and 5-aminovaleramide amidohydrolase (DavA) inserted into its genome to form <em>H. bluephagenesis</em> YF117Δ<em>gabT</em><sub>1+2</sub>, which produced 16.4 g L<sup>−1</sup> and 67.4 g L<sup>−1</sup> 5-AVA in flask cultures and in 7 L bioreactor, respectively. It was able to <em>de novo</em> synthesize 5-AVA from glucose by L-lysine-overproducing <em>H. bluephagenesis</em> TD226. Corn steep liquor was used instead of yeast extract for cost reduction during the 5-AVA production. Using promoter engineering based on P<sub><em>porin</em></sub> mutant library for downstream genes, <em>H. bluephagenesis</em> YF117 harboring pSEVA341-P<sub><em>porin</em></sub> <sub>42</sub>-<em>yqhD</em><sub>EC</sub> produced 6 g L<sup>−1</sup> 5HV in shake flask growth<em>,</em> while <em>H. bluephagenesis</em> YF117 harboring pSEVA341-P<sub><em>porin</em></sub> <sub>42</sub>-<em>yqhD</em><sub>EC</sub>-P<sub><em>porin</em></sub> <sub>278</sub>-<em>phaC</em><sub>RE</sub><em>-abfT</em> synthesized 42 wt% P(3HB-<em>co</em>-4.8 mol% 5HV) under the same condition. Thus, <em>H. bluephagenesis</em> was successfully engineered to produce 5-AVA and 5HV in supernatant and intracellular P(3HB-<em>co</em>-5HV) utilizing L-lysine as the substrate.</p></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":null,"pages":null},"PeriodicalIF":6.8000,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S109671762300174X/pdfft?md5=fd9c929412002187037a7287bdf5f410&pid=1-s2.0-S109671762300174X-main.pdf","citationCount":"0","resultStr":"{\"title\":\"Metabolic engineering of Halomonas bluephagenesis for production of five carbon molecular chemicals derived from L-lysine\",\"authors\":\"Fang Yang , Huan Wang , Cuihuan Zhao , Lizhan Zhang , Xu Liu , Helen Park , Yiping Yuan , Jian-Wen Ye , Qiong Wu , Guo-Qiang Chen\",\"doi\":\"10.1016/j.ymben.2023.12.001\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>5-Aminovaleric acid (5-AVA), 5-hydroxyvalerate (5HV), copolymer P(3HB-<em>co</em>-5HV) of <em>3</em>-hydroxybutyrate (3HB) and 5HV were produced from L-lysine as a substrate by recombinant <em>Halomonas bluephagenesis</em> constructed based on codon optimization, deletions of competitive pathway and L-lysine export protein, and three copies of <em>davBA</em> genes encoding L-lysine monooxygenase (DavB) and 5-aminovaleramide amidohydrolase (DavA) inserted into its genome to form <em>H. bluephagenesis</em> YF117Δ<em>gabT</em><sub>1+2</sub>, which produced 16.4 g L<sup>−1</sup> and 67.4 g L<sup>−1</sup> 5-AVA in flask cultures and in 7 L bioreactor, respectively. It was able to <em>de novo</em> synthesize 5-AVA from glucose by L-lysine-overproducing <em>H. bluephagenesis</em> TD226. Corn steep liquor was used instead of yeast extract for cost reduction during the 5-AVA production. Using promoter engineering based on P<sub><em>porin</em></sub> mutant library for downstream genes, <em>H. bluephagenesis</em> YF117 harboring pSEVA341-P<sub><em>porin</em></sub> <sub>42</sub>-<em>yqhD</em><sub>EC</sub> produced 6 g L<sup>−1</sup> 5HV in shake flask growth<em>,</em> while <em>H. bluephagenesis</em> YF117 harboring pSEVA341-P<sub><em>porin</em></sub> <sub>42</sub>-<em>yqhD</em><sub>EC</sub>-P<sub><em>porin</em></sub> <sub>278</sub>-<em>phaC</em><sub>RE</sub><em>-abfT</em> synthesized 42 wt% P(3HB-<em>co</em>-4.8 mol% 5HV) under the same condition. Thus, <em>H. bluephagenesis</em> was successfully engineered to produce 5-AVA and 5HV in supernatant and intracellular P(3HB-<em>co</em>-5HV) utilizing L-lysine as the substrate.</p></div>\",\"PeriodicalId\":18483,\"journal\":{\"name\":\"Metabolic engineering\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":6.8000,\"publicationDate\":\"2024-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S109671762300174X/pdfft?md5=fd9c929412002187037a7287bdf5f410&pid=1-s2.0-S109671762300174X-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Metabolic engineering\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S109671762300174X\",\"RegionNum\":1,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Metabolic engineering","FirstCategoryId":"5","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S109671762300174X","RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
Metabolic engineering of Halomonas bluephagenesis for production of five carbon molecular chemicals derived from L-lysine
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 YF117ΔgabT1+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 Pporin mutant library for downstream genes, H. bluephagenesis YF117 harboring pSEVA341-Pporin42-yqhDEC produced 6 g L−1 5HV in shake flask growth, while H. bluephagenesis YF117 harboring pSEVA341-Pporin42-yqhDEC-Pporin278-phaCRE-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.
期刊介绍:
Metabolic Engineering (MBE) is a journal that focuses on publishing original research papers on the directed modulation of metabolic pathways for metabolite overproduction or the enhancement of cellular properties. It welcomes papers that describe the engineering of native pathways and the synthesis of heterologous pathways to convert microorganisms into microbial cell factories. The journal covers experimental, computational, and modeling approaches for understanding metabolic pathways and manipulating them through genetic, media, or environmental means. Effective exploration of metabolic pathways necessitates the use of molecular biology and biochemistry methods, as well as engineering techniques for modeling and data analysis. MBE serves as a platform for interdisciplinary research in fields such as biochemistry, molecular biology, applied microbiology, cellular physiology, cellular nutrition in health and disease, and biochemical engineering. The journal publishes various types of papers, including original research papers and review papers. It is indexed and abstracted in databases such as Scopus, Embase, EMBiology, Current Contents - Life Sciences and Clinical Medicine, Science Citation Index, PubMed/Medline, CAS and Biotechnology Citation Index.