Enzymatic synthesis of nylon precursors by 4-aminobutyrate aminotransferase and 6-oxohexanoate dehydrogenase

IF 2.5 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Hoe-Suk Lee, Yung-Hun Yang, Young Joo Yeon, Hyun June Park
{"title":"Enzymatic synthesis of nylon precursors by 4-aminobutyrate aminotransferase and 6-oxohexanoate dehydrogenase","authors":"Hoe-Suk Lee, Yung-Hun Yang, Young Joo Yeon, Hyun June Park","doi":"10.1007/s12257-024-00011-x","DOIUrl":null,"url":null,"abstract":"<p>6-Aminocaproic acid and adipic acid are the key value-added chemical precursors in the pharmaceutical, solvent and polyamide industry, including nylon-6, and nylon-6, 6. An enzymatic interconversion of the two precursors can provide a convenient and eco-friendly biosynthetic route to each of the precursors and thus, require analysis of the reaction process. Herein, an in vitro enzymatic method was employed to convert the two precursors while most studies so far have focused on the whole cell bioconversion to investigate the process. 4-Aminobutyrate aminotransferase was utilized to mediate the reactions between 6-aminocaproic acid and the intermediate 6-oxohexanoic acid with the aid of pyridoxal 5’-phosphate and amine donor/acceptor. 6-Oxohexanoate dehydrogenase was utilized for the reaction from 6-oxohexanoic acid to adipic acid with NADP<sup>+</sup>. A range of reaction conditions were investigated including the type of amine donor, pH conditions, the concentrations of enzyme and amine donor/acceptor. The optimum condition resulted in 78% yield for the reaction from 6-oxohexanoic acid to 6-aminocaproic acid. The yield for the one-pot, two-step enzymatic cascade from 6-aminocaproic acid via 6-oxohexanoate intermediate to adipic acid was 88%, which was higher than the yield for each individual step in the cascade, with 31% and 32%, for the first and second step, respectively. Furthermore, structural analysis on the active site of the 4-aminobutyrate aminotransferase docked with a range of amine donors implicates the optimal donor is glutamate in accordance with the experimental data and suggests enzyme engineering possibilities for more readily available donors to facilitate the industrial application of the process.</p>","PeriodicalId":8936,"journal":{"name":"Biotechnology and Bioprocess Engineering","volume":"9 1","pages":""},"PeriodicalIF":2.5000,"publicationDate":"2024-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biotechnology and Bioprocess Engineering","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1007/s12257-024-00011-x","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0

Abstract

6-Aminocaproic acid and adipic acid are the key value-added chemical precursors in the pharmaceutical, solvent and polyamide industry, including nylon-6, and nylon-6, 6. An enzymatic interconversion of the two precursors can provide a convenient and eco-friendly biosynthetic route to each of the precursors and thus, require analysis of the reaction process. Herein, an in vitro enzymatic method was employed to convert the two precursors while most studies so far have focused on the whole cell bioconversion to investigate the process. 4-Aminobutyrate aminotransferase was utilized to mediate the reactions between 6-aminocaproic acid and the intermediate 6-oxohexanoic acid with the aid of pyridoxal 5’-phosphate and amine donor/acceptor. 6-Oxohexanoate dehydrogenase was utilized for the reaction from 6-oxohexanoic acid to adipic acid with NADP+. A range of reaction conditions were investigated including the type of amine donor, pH conditions, the concentrations of enzyme and amine donor/acceptor. The optimum condition resulted in 78% yield for the reaction from 6-oxohexanoic acid to 6-aminocaproic acid. The yield for the one-pot, two-step enzymatic cascade from 6-aminocaproic acid via 6-oxohexanoate intermediate to adipic acid was 88%, which was higher than the yield for each individual step in the cascade, with 31% and 32%, for the first and second step, respectively. Furthermore, structural analysis on the active site of the 4-aminobutyrate aminotransferase docked with a range of amine donors implicates the optimal donor is glutamate in accordance with the experimental data and suggests enzyme engineering possibilities for more readily available donors to facilitate the industrial application of the process.

Abstract Image

4-氨基丁酸氨基转移酶和 6-氧代己酸脱氢酶对尼龙前体的酶促合成作用
6-氨基己酸和己二酸是医药、溶剂和聚酰胺行业(包括尼龙-6 和尼龙-6, 6)的关键增值化学前体。这两种前体的酶促相互转化可为每种前体提供一种方便、环保的生物合成途径,因此需要对反应过程进行分析。迄今为止,大多数研究都侧重于全细胞生物转化来研究这一过程,而本文则采用体外酶法来转化这两种前体。在 5'-磷酸吡哆醛和胺供体/受体的帮助下,4-氨基丁酸氨基转移酶被用来介导 6-氨基己酸和中间体 6-氧代己酸之间的反应。在 6-oxohexanoic acid 与 NADP+ 的反应中,使用了 6-oxohexanoic acid 脱氢酶。研究了一系列反应条件,包括胺供体的类型、pH 值条件、酶和胺供体/受体的浓度。在最佳条件下,从 6-氧代己酸到 6-氨基己酸的反应产率为 78%。通过 6-oxohexanoate 中间体从 6-aminocaproic acid 到己二酸的一锅两步酶促级联反应的产率为 88%,高于级联反应中每个步骤的产率,第一步和第二步的产率分别为 31% 和 32%。此外,对与一系列胺供体对接的 4-氨基丁酸氨基转移酶活性位点的结构分析表明,最佳供体是谷氨酸,这与实验数据相符,并提出了酶工程的可能性,以获得更多容易获得的供体,从而促进该过程的工业应用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Biotechnology and Bioprocess Engineering
Biotechnology and Bioprocess Engineering 工程技术-生物工程与应用微生物
CiteScore
5.00
自引率
12.50%
发文量
79
审稿时长
3 months
期刊介绍: Biotechnology and Bioprocess Engineering is an international bimonthly journal published by the Korean Society for Biotechnology and Bioengineering. BBE is devoted to the advancement in science and technology in the wide area of biotechnology, bioengineering, and (bio)medical engineering. This includes but is not limited to applied molecular and cell biology, engineered biocatalysis and biotransformation, metabolic engineering and systems biology, bioseparation and bioprocess engineering, cell culture technology, environmental and food biotechnology, pharmaceutics and biopharmaceutics, biomaterials engineering, nanobiotechnology, and biosensor and bioelectronics.
×
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