酿酒酵母中 NADPH 供应策略的比较分析:以从二木糖生产二木糖醇为例进行研究

IF 3.7 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Metabolic Engineering Communications Pub Date : 2024-07-05 DOI:10.1016/j.mec.2024.e00245
Priti Regmi , Melanie Knesebeck , Eckhard Boles , Dirk Weuster-Botz , Mislav Oreb
{"title":"酿酒酵母中 NADPH 供应策略的比较分析:以从二木糖生产二木糖醇为例进行研究","authors":"Priti Regmi ,&nbsp;Melanie Knesebeck ,&nbsp;Eckhard Boles ,&nbsp;Dirk Weuster-Botz ,&nbsp;Mislav Oreb","doi":"10.1016/j.mec.2024.e00245","DOIUrl":null,"url":null,"abstract":"<div><p>Enhancing the supply of the redox cofactor NADPH in metabolically engineered cells is a critical target for optimizing the synthesis of many product classes, such as fatty acids or terpenoids. In <em>S. cerevisiae</em>, several successful approaches have been developed in different experimental contexts. However, their systematic comparison has not been reported. Here, we established the reduction of xylose to xylitol by an NADPH-dependent xylose reductase as a model reaction to compare the efficacy of different NADPH supply strategies in the course of a batch fermentation, in which glucose and ethanol are sequentially used as carbon sources and redox donors. We show that strains overexpressing the glucose-6-phosphate dehydrogenase Zwf1 perform best, producing up to 16.9 g L<sup>−1</sup> xylitol from 20 g L<sup>−1</sup> xylose in stirred tank bioreactors. The beneficial effect of increased Zwf1 activity is especially pronounced during the ethanol consumption phase. The same notion applies to the deletion of the aldehyde dehydrogenase <em>ALD6</em> gene, albeit at a quantitatively lower level. Reduced expression of the phosphoglucose isomerase Pgi1 and heterologous expression of the NADP<sup>+</sup>-dependent glyceraldehyde-3-phosphate dehydrogenase Gdp1 from <em>Kluyveromyces lactis</em> acted synergistically with <em>ZWF1</em> overexpression in the presence of glucose, but had a detrimental effect after the diauxic shift. Expression of the mitochondrial NADH kinase Pos5 in the cytosol likewise improved the production of xylitol only on glucose, but not in combination with enhanced Zwf1 activity. To demonstrate the generalizability of our observations, we show that the most promising strategies – <em>ZWF1</em> overexpression and deletion of <em>ALD6</em> - also improve the production of <span>l</span>-galactonate from <span>d</span>-galacturonic acid. Therefore, we expect that these findings will provide valuable guidelines for engineering not only the production of xylitol but also of diverse other pathways that require NADPH.</p></div>","PeriodicalId":18695,"journal":{"name":"Metabolic Engineering Communications","volume":null,"pages":null},"PeriodicalIF":3.7000,"publicationDate":"2024-07-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2214030124000142/pdfft?md5=7ade0b7c412cf8487310e2ebc5404b91&pid=1-s2.0-S2214030124000142-main.pdf","citationCount":"0","resultStr":"{\"title\":\"A comparative analysis of NADPH supply strategies in Saccharomyces cerevisiae: Production of d-xylitol from d-xylose as a case study\",\"authors\":\"Priti Regmi ,&nbsp;Melanie Knesebeck ,&nbsp;Eckhard Boles ,&nbsp;Dirk Weuster-Botz ,&nbsp;Mislav Oreb\",\"doi\":\"10.1016/j.mec.2024.e00245\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Enhancing the supply of the redox cofactor NADPH in metabolically engineered cells is a critical target for optimizing the synthesis of many product classes, such as fatty acids or terpenoids. In <em>S. cerevisiae</em>, several successful approaches have been developed in different experimental contexts. However, their systematic comparison has not been reported. Here, we established the reduction of xylose to xylitol by an NADPH-dependent xylose reductase as a model reaction to compare the efficacy of different NADPH supply strategies in the course of a batch fermentation, in which glucose and ethanol are sequentially used as carbon sources and redox donors. We show that strains overexpressing the glucose-6-phosphate dehydrogenase Zwf1 perform best, producing up to 16.9 g L<sup>−1</sup> xylitol from 20 g L<sup>−1</sup> xylose in stirred tank bioreactors. The beneficial effect of increased Zwf1 activity is especially pronounced during the ethanol consumption phase. The same notion applies to the deletion of the aldehyde dehydrogenase <em>ALD6</em> gene, albeit at a quantitatively lower level. Reduced expression of the phosphoglucose isomerase Pgi1 and heterologous expression of the NADP<sup>+</sup>-dependent glyceraldehyde-3-phosphate dehydrogenase Gdp1 from <em>Kluyveromyces lactis</em> acted synergistically with <em>ZWF1</em> overexpression in the presence of glucose, but had a detrimental effect after the diauxic shift. Expression of the mitochondrial NADH kinase Pos5 in the cytosol likewise improved the production of xylitol only on glucose, but not in combination with enhanced Zwf1 activity. To demonstrate the generalizability of our observations, we show that the most promising strategies – <em>ZWF1</em> overexpression and deletion of <em>ALD6</em> - also improve the production of <span>l</span>-galactonate from <span>d</span>-galacturonic acid. Therefore, we expect that these findings will provide valuable guidelines for engineering not only the production of xylitol but also of diverse other pathways that require NADPH.</p></div>\",\"PeriodicalId\":18695,\"journal\":{\"name\":\"Metabolic Engineering Communications\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":3.7000,\"publicationDate\":\"2024-07-05\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S2214030124000142/pdfft?md5=7ade0b7c412cf8487310e2ebc5404b91&pid=1-s2.0-S2214030124000142-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Metabolic Engineering Communications\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2214030124000142\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Metabolic Engineering Communications","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2214030124000142","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0

摘要

提高代谢工程细胞中氧化还原辅助因子 NADPH 的供应量是优化脂肪酸或萜类化合物等多种产品合成的关键目标。在 S. cerevisiae 中,人们在不同的实验环境中开发出了几种成功的方法。但是,还没有对这些方法进行系统比较的报道。在这里,我们将依赖 NADPH 的木糖还原酶将木糖还原成木糖醇的反应作为一个模型反应,以比较在批量发酵过程中不同 NADPH 供应策略的功效,在批量发酵过程中,葡萄糖和乙醇依次被用作碳源和氧化还原供体。我们的研究表明,过量表达葡萄糖-6-磷酸脱氢酶 Zwf1 的菌株表现最佳,在搅拌罐生物反应器中可从 20 克/升木糖中产生高达 16.9 克/升的木糖醇。在乙醇消耗阶段,Zwf1 活性增加的有利影响尤其明显。醛脱氢酶 ALD6 基因的缺失也是同样的道理,尽管其数量水平较低。磷酸葡萄糖异构酶 Pgi1 的表达量减少,以及来自乳酸克鲁维酵母菌的依赖 NADP+ 的甘油醛-3-磷酸脱氢酶 Gdp1 的异源表达,在葡萄糖存在的情况下与 ZWF1 的过量表达具有协同作用,但在二重转移后则产生了不利影响。同样,在细胞质中表达线粒体 NADH 激酶 Pos5 也只能在葡萄糖条件下提高木糖醇的产量,但不能与 Zwf1 活性的增强相结合。为了证明我们的观察结果具有普遍性,我们发现最有前景的策略--ZWF1 的过表达和 ALD6 的缺失--也能改善由 d-半乳糖醛酸产生 l-半乳糖醛酸的情况。因此,我们希望这些发现不仅能为木糖醇的生产工程提供有价值的指导,也能为需要 NADPH 的其他各种途径的生产工程提供有价值的指导。
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
A comparative analysis of NADPH supply strategies in Saccharomyces cerevisiae: Production of d-xylitol from d-xylose as a case study

Enhancing the supply of the redox cofactor NADPH in metabolically engineered cells is a critical target for optimizing the synthesis of many product classes, such as fatty acids or terpenoids. In S. cerevisiae, several successful approaches have been developed in different experimental contexts. However, their systematic comparison has not been reported. Here, we established the reduction of xylose to xylitol by an NADPH-dependent xylose reductase as a model reaction to compare the efficacy of different NADPH supply strategies in the course of a batch fermentation, in which glucose and ethanol are sequentially used as carbon sources and redox donors. We show that strains overexpressing the glucose-6-phosphate dehydrogenase Zwf1 perform best, producing up to 16.9 g L−1 xylitol from 20 g L−1 xylose in stirred tank bioreactors. The beneficial effect of increased Zwf1 activity is especially pronounced during the ethanol consumption phase. The same notion applies to the deletion of the aldehyde dehydrogenase ALD6 gene, albeit at a quantitatively lower level. Reduced expression of the phosphoglucose isomerase Pgi1 and heterologous expression of the NADP+-dependent glyceraldehyde-3-phosphate dehydrogenase Gdp1 from Kluyveromyces lactis acted synergistically with ZWF1 overexpression in the presence of glucose, but had a detrimental effect after the diauxic shift. Expression of the mitochondrial NADH kinase Pos5 in the cytosol likewise improved the production of xylitol only on glucose, but not in combination with enhanced Zwf1 activity. To demonstrate the generalizability of our observations, we show that the most promising strategies – ZWF1 overexpression and deletion of ALD6 - also improve the production of l-galactonate from d-galacturonic acid. Therefore, we expect that these findings will provide valuable guidelines for engineering not only the production of xylitol but also of diverse other pathways that require NADPH.

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
Metabolic Engineering Communications
Metabolic Engineering Communications Medicine-Endocrinology, Diabetes and Metabolism
CiteScore
13.30
自引率
1.90%
发文量
22
审稿时长
18 weeks
期刊介绍: Metabolic Engineering Communications, a companion title to Metabolic Engineering (MBE), is devoted to publishing original research in the areas of metabolic engineering, synthetic biology, computational biology and systems biology for problems related to metabolism and the engineering of metabolism for the production of fuels, chemicals, and pharmaceuticals. The journal will carry articles on the design, construction, and analysis of biological systems ranging from pathway components to biological complexes and genomes (including genomic, analytical and bioinformatics methods) in suitable host cells to allow them to produce novel compounds of industrial and medical interest. Demonstrations of regulatory designs and synthetic circuits that alter the performance of biochemical pathways and cellular processes will also be presented. Metabolic Engineering Communications complements MBE by publishing articles that are either shorter than those published in the full journal, or which describe key elements of larger metabolic engineering efforts.
期刊最新文献
Metabolic engineering of Acinetobacter baylyi ADP1 for naringenin production PEZy-miner: An artificial intelligence driven approach for the discovery of plastic-degrading enzyme candidates Production of (R)-citramalate by engineered Saccharomyces cerevisiae Engineering thioesterase as a driving force for novel itaconate production via its degradation scheme A comparative analysis of NADPH supply strategies in Saccharomyces cerevisiae: Production of d-xylitol from d-xylose as a case study
×
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