Xin Wang , Peipei Cai , Kequan Chen, Pingkai Ouyang
{"title":"利用工程大肠杆菌全细胞生物催化剂从赖氨酸高效生产5-氨基戊酸酯","authors":"Xin Wang , Peipei Cai , Kequan Chen, Pingkai Ouyang","doi":"10.1016/j.molcatb.2016.10.008","DOIUrl":null,"url":null,"abstract":"<div><p>In this study, we developed a whole-cell biocatalysis process for high-level conversion of <span>l</span>-lysine into 5-aminovalerate. To obtain the highly efficient whole-cell biocatalyst, five expression plasmids were constructed to optimize the expression of 5-aminovaleramide amidohydrolase and <span>l</span>-lysine 2-monooxygenase in <em>Escherichia coli</em>. The engineered strain BL-22A-RB-YB harboring plasmid pET22b-davA, pRSFDuet-davB and pACYCDuet-davB was correspondingly obtained. Subsequently, the effects of induction conditions, reaction temperature, metal ion additives, and cell permeability on the whole-cell biocatalyst system were evaluated to improve biocatalytic efficiency. Under optimized reaction conditions, 95.3<!--> <!-->g/L 5-aminovalerate was synthesized from 120<!--> <!-->g/L <span>l</span>-lysine with a yield of 99.1%, and 103.1<!--> <!-->g/L 5-aminovalerate was produced from 150<!--> <!-->g/L <span>l</span>-lysine with a molar yield of 85.7%. The 5-aminovalerate production was then further improved using a <span>l</span>-lysine fed-batch strategy, and a hyper 5-aminovalerate production of 240.7<!--> <!-->g/L was achieved within 28<!--> <!-->h with a yield of 86.8%. The whole-cell biocatalytic system described here demonstrated an environmentally friendly strategy for industrial production of 5-aminovalerate.</p></div>","PeriodicalId":16416,"journal":{"name":"Journal of Molecular Catalysis B-enzymatic","volume":"134 ","pages":"Pages 115-121"},"PeriodicalIF":0.0000,"publicationDate":"2016-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.molcatb.2016.10.008","citationCount":"24","resultStr":"{\"title\":\"Efficient production of 5-aminovalerate from l-lysine by engineered Escherichia coli whole-cell biocatalysts\",\"authors\":\"Xin Wang , Peipei Cai , Kequan Chen, Pingkai Ouyang\",\"doi\":\"10.1016/j.molcatb.2016.10.008\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>In this study, we developed a whole-cell biocatalysis process for high-level conversion of <span>l</span>-lysine into 5-aminovalerate. To obtain the highly efficient whole-cell biocatalyst, five expression plasmids were constructed to optimize the expression of 5-aminovaleramide amidohydrolase and <span>l</span>-lysine 2-monooxygenase in <em>Escherichia coli</em>. The engineered strain BL-22A-RB-YB harboring plasmid pET22b-davA, pRSFDuet-davB and pACYCDuet-davB was correspondingly obtained. Subsequently, the effects of induction conditions, reaction temperature, metal ion additives, and cell permeability on the whole-cell biocatalyst system were evaluated to improve biocatalytic efficiency. Under optimized reaction conditions, 95.3<!--> <!-->g/L 5-aminovalerate was synthesized from 120<!--> <!-->g/L <span>l</span>-lysine with a yield of 99.1%, and 103.1<!--> <!-->g/L 5-aminovalerate was produced from 150<!--> <!-->g/L <span>l</span>-lysine with a molar yield of 85.7%. The 5-aminovalerate production was then further improved using a <span>l</span>-lysine fed-batch strategy, and a hyper 5-aminovalerate production of 240.7<!--> <!-->g/L was achieved within 28<!--> <!-->h with a yield of 86.8%. The whole-cell biocatalytic system described here demonstrated an environmentally friendly strategy for industrial production of 5-aminovalerate.</p></div>\",\"PeriodicalId\":16416,\"journal\":{\"name\":\"Journal of Molecular Catalysis B-enzymatic\",\"volume\":\"134 \",\"pages\":\"Pages 115-121\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2016-12-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/j.molcatb.2016.10.008\",\"citationCount\":\"24\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Molecular Catalysis B-enzymatic\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1381117716302016\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"Chemical Engineering\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Molecular Catalysis B-enzymatic","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1381117716302016","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"Chemical Engineering","Score":null,"Total":0}
Efficient production of 5-aminovalerate from l-lysine by engineered Escherichia coli whole-cell biocatalysts
In this study, we developed a whole-cell biocatalysis process for high-level conversion of l-lysine into 5-aminovalerate. To obtain the highly efficient whole-cell biocatalyst, five expression plasmids were constructed to optimize the expression of 5-aminovaleramide amidohydrolase and l-lysine 2-monooxygenase in Escherichia coli. The engineered strain BL-22A-RB-YB harboring plasmid pET22b-davA, pRSFDuet-davB and pACYCDuet-davB was correspondingly obtained. Subsequently, the effects of induction conditions, reaction temperature, metal ion additives, and cell permeability on the whole-cell biocatalyst system were evaluated to improve biocatalytic efficiency. Under optimized reaction conditions, 95.3 g/L 5-aminovalerate was synthesized from 120 g/L l-lysine with a yield of 99.1%, and 103.1 g/L 5-aminovalerate was produced from 150 g/L l-lysine with a molar yield of 85.7%. The 5-aminovalerate production was then further improved using a l-lysine fed-batch strategy, and a hyper 5-aminovalerate production of 240.7 g/L was achieved within 28 h with a yield of 86.8%. The whole-cell biocatalytic system described here demonstrated an environmentally friendly strategy for industrial production of 5-aminovalerate.
期刊介绍:
Journal of Molecular Catalysis B: Enzymatic is an international forum for researchers and product developers in the applications of whole-cell and cell-free enzymes as catalysts in organic synthesis. Emphasis is on mechanistic and synthetic aspects of the biocatalytic transformation.
Papers should report novel and significant advances in one or more of the following topics;
Applied and fundamental studies of enzymes used for biocatalysis;
Industrial applications of enzymatic processes, e.g. in fine chemical synthesis;
Chemo-, regio- and enantioselective transformations;
Screening for biocatalysts;
Integration of biocatalytic and chemical steps in organic syntheses;
Novel biocatalysts, e.g. enzymes from extremophiles and catalytic antibodies;
Enzyme immobilization and stabilization, particularly in non-conventional media;
Bioprocess engineering aspects, e.g. membrane bioreactors;
Improvement of catalytic performance of enzymes, e.g. by protein engineering or chemical modification;
Structural studies, including computer simulation, relating to substrate specificity and reaction selectivity;
Biomimetic studies related to enzymatic transformations.