基于渗透酿酒酵母细胞中人类 CYP2C9 酶活性的逻辑遗传酶门的设计和工程学研究

IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Synthetic and Systems Biotechnology Pub Date : 2024-03-29 DOI:10.1016/j.synbio.2024.03.013
Rana Azeem Ashraf, Matthias Bureik, Mario Andrea Marchisio
{"title":"基于渗透酿酒酵母细胞中人类 CYP2C9 酶活性的逻辑遗传酶门的设计和工程学研究","authors":"Rana Azeem Ashraf,&nbsp;Matthias Bureik,&nbsp;Mario Andrea Marchisio","doi":"10.1016/j.synbio.2024.03.013","DOIUrl":null,"url":null,"abstract":"<div><p>Gene circuits allow cells to carry out complex functions such as the precise regulation of biological metabolic processes. In this study, we combined, in the yeast <em>S. cerevisiae</em>, genetic regulatory elements with the enzymatic reactions of the human CYP2C9 and its redox partner CPR on luciferin substrates and diclofenac. <em>S. cerevisiae</em> cells were permeabilized and used as enzyme bags in order to host these metabolic reactions. We engineered three different (genetic)-enzymatic basic Boolean gates (YES, NOT, and N-IMPLY). In the YES and N-IMPLY gates, human CYP2C9 was expressed under the galactose-inducible <em>GAL1</em> promoter. The carbon monoxide releasing molecule CORM-401 was used as an input in the NOT and N-IMPLY gates to impair CYP2C9 activity through inhibition of the Fe<sup>+2</sup>- heme prosthetic group in the active site of the human enzyme. Our study provides a new approach in designing synthetic bio-circuits and optimizing experimental conditions to favor the heterologous expression of human drug metabolic enzymes over their endogenous counterparts. This new approach will help study precise metabolic attributes of human P450s.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"9 3","pages":"Pages 406-415"},"PeriodicalIF":4.4000,"publicationDate":"2024-03-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000474/pdfft?md5=1b0e86900ca2e71dc42b1d5d44a1c2c4&pid=1-s2.0-S2405805X24000474-main.pdf","citationCount":"0","resultStr":"{\"title\":\"Design and engineering of logic genetic-enzymatic gates based on the activity of the human CYP2C9 enzyme in permeabilized Saccharomyces cerevisiae cells\",\"authors\":\"Rana Azeem Ashraf,&nbsp;Matthias Bureik,&nbsp;Mario Andrea Marchisio\",\"doi\":\"10.1016/j.synbio.2024.03.013\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Gene circuits allow cells to carry out complex functions such as the precise regulation of biological metabolic processes. In this study, we combined, in the yeast <em>S. cerevisiae</em>, genetic regulatory elements with the enzymatic reactions of the human CYP2C9 and its redox partner CPR on luciferin substrates and diclofenac. <em>S. cerevisiae</em> cells were permeabilized and used as enzyme bags in order to host these metabolic reactions. We engineered three different (genetic)-enzymatic basic Boolean gates (YES, NOT, and N-IMPLY). In the YES and N-IMPLY gates, human CYP2C9 was expressed under the galactose-inducible <em>GAL1</em> promoter. The carbon monoxide releasing molecule CORM-401 was used as an input in the NOT and N-IMPLY gates to impair CYP2C9 activity through inhibition of the Fe<sup>+2</sup>- heme prosthetic group in the active site of the human enzyme. Our study provides a new approach in designing synthetic bio-circuits and optimizing experimental conditions to favor the heterologous expression of human drug metabolic enzymes over their endogenous counterparts. This new approach will help study precise metabolic attributes of human P450s.</p></div>\",\"PeriodicalId\":22148,\"journal\":{\"name\":\"Synthetic and Systems Biotechnology\",\"volume\":\"9 3\",\"pages\":\"Pages 406-415\"},\"PeriodicalIF\":4.4000,\"publicationDate\":\"2024-03-29\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S2405805X24000474/pdfft?md5=1b0e86900ca2e71dc42b1d5d44a1c2c4&pid=1-s2.0-S2405805X24000474-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Synthetic and Systems Biotechnology\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2405805X24000474\",\"RegionNum\":2,\"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":"Synthetic and Systems Biotechnology","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2405805X24000474","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0

摘要

基因回路使细胞能够执行复杂的功能,如精确调节生物代谢过程。在这项研究中,我们在酵母 S. cerevisiae 中将基因调控元件与人类 CYP2C9 及其氧化还原伙伴 CPR 对荧光素底物和双氯芬酸的酶促反应结合起来。对 S. cerevisiae 细胞进行了渗透处理,并将其用作酶袋,以承载这些代谢反应。我们设计了三种不同的(遗传)酶基本布尔门(YES、NOT 和 N-IMPLY)。在 YES 和 N-IMPLY 门中,人 CYP2C9 在半乳糖诱导的 GAL1 启动子下表达。在 NOT 和 N-IMPLY 门中,一氧化碳释放分子 CORM-401 被用作输入,通过抑制人类酶活性位点中的 Fe+2- 血红素修复基团来损害 CYP2C9 的活性。我们的研究为设计合成生物电路和优化实验条件提供了一种新方法,使人类药物代谢酶的异源表达优于其内源性对应物。这种新方法将有助于研究人类 P450s 的精确代谢属性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
Design and engineering of logic genetic-enzymatic gates based on the activity of the human CYP2C9 enzyme in permeabilized Saccharomyces cerevisiae cells

Gene circuits allow cells to carry out complex functions such as the precise regulation of biological metabolic processes. In this study, we combined, in the yeast S. cerevisiae, genetic regulatory elements with the enzymatic reactions of the human CYP2C9 and its redox partner CPR on luciferin substrates and diclofenac. S. cerevisiae cells were permeabilized and used as enzyme bags in order to host these metabolic reactions. We engineered three different (genetic)-enzymatic basic Boolean gates (YES, NOT, and N-IMPLY). In the YES and N-IMPLY gates, human CYP2C9 was expressed under the galactose-inducible GAL1 promoter. The carbon monoxide releasing molecule CORM-401 was used as an input in the NOT and N-IMPLY gates to impair CYP2C9 activity through inhibition of the Fe+2- heme prosthetic group in the active site of the human enzyme. Our study provides a new approach in designing synthetic bio-circuits and optimizing experimental conditions to favor the heterologous expression of human drug metabolic enzymes over their endogenous counterparts. This new approach will help study precise metabolic attributes of human P450s.

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
Synthetic and Systems Biotechnology
Synthetic and Systems Biotechnology BIOTECHNOLOGY & APPLIED MICROBIOLOGY-
CiteScore
6.90
自引率
12.50%
发文量
90
审稿时长
67 days
期刊介绍: Synthetic and Systems Biotechnology aims to promote the communication of original research in synthetic and systems biology, with strong emphasis on applications towards biotechnology. This journal is a quarterly peer-reviewed journal led by Editor-in-Chief Lixin Zhang. The journal publishes high-quality research; focusing on integrative approaches to enable the understanding and design of biological systems, and research to develop the application of systems and synthetic biology to natural systems. This journal will publish Articles, Short notes, Methods, Mini Reviews, Commentary and Conference reviews.
期刊最新文献
Semi-rational design and modification of phosphoketolase to improve the yield of tyrosol in Saccharomyces cerevisiae Pathway and protein channel engineering of Bacillus subtilis for improved production of desthiobiotin and biotin Harnessing microbial heterogeneity for improved biosynthesis fueled by synthetic biology Recent advances in the biosynthesis and production optimization of gentamicin: A critical review Coordinated regulation of two LacI family regulators, GvmR and GvmR2, on guvermectin production in Streptomyces caniferus
×
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
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 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 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1