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

Q2 Chemical Engineering
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 ,&nbsp;Peipei Cai ,&nbsp;Kequan Chen,&nbsp;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 ,&nbsp;Peipei Cai ,&nbsp;Kequan Chen,&nbsp;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}
引用次数: 24

摘要

在这项研究中,我们开发了一种全细胞生物催化工艺,用于将l-赖氨酸高水平转化为5-氨基戊酸盐。为了获得高效的全细胞生物催化剂,构建了5个表达质粒,优化了5-氨基戊酰胺酰胺水解酶和l-赖氨酸2-单加氧酶在大肠杆菌中的表达。相应获得含有质粒pET22b-davA、pRSFDuet-davB和pACYCDuet-davB的工程菌株BL-22A-RB-YB。随后,研究了诱导条件、反应温度、金属离子添加剂和细胞通透性对全细胞生物催化剂体系的影响,以提高生物催化效率。在优化的反应条件下,120 g/L赖氨酸可合成95.3 g/L 5-氨基戊酸酯,产率为99.1%;150 g/L赖氨酸可合成103.1 g/L 5-氨基戊酸酯,摩尔产率为85.7%。采用L -赖氨酸补料分批策略进一步提高5-氨基戊酸酯的产量,在28 h内达到240.7 g/L的超5-氨基戊酸酯产量,产量为86.8%。这里描述的全细胞生物催化系统展示了工业生产5-氨基戊酸酯的环保策略。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Efficient production of 5-aminovalerate from l-lysine by engineered Escherichia coli whole-cell biocatalysts

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.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
Journal of Molecular Catalysis B-enzymatic
Journal of Molecular Catalysis B-enzymatic 生物-生化与分子生物学
CiteScore
2.58
自引率
0.00%
发文量
0
审稿时长
3.4 months
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.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信