Characterisation and Harnessing of 5-Hydroxymethylfurfural Metabolism in Pseudomonas umsongensis GO16 for the Production of 2,5-Furandicarboxylic Acid

IF 5.7 2区 生物学
Rhys Orimaco, Pauric Donnelly, Seán Sexton, Aoife McLoughlin, Sophie Kelly, Kevin E. O'Connor, Nick Wierckx, Tanja Narančić
{"title":"Characterisation and Harnessing of 5-Hydroxymethylfurfural Metabolism in Pseudomonas umsongensis GO16 for the Production of 2,5-Furandicarboxylic Acid","authors":"Rhys Orimaco,&nbsp;Pauric Donnelly,&nbsp;Seán Sexton,&nbsp;Aoife McLoughlin,&nbsp;Sophie Kelly,&nbsp;Kevin E. O'Connor,&nbsp;Nick Wierckx,&nbsp;Tanja Narančić","doi":"10.1111/1751-7915.70159","DOIUrl":null,"url":null,"abstract":"<p>In the search for biobased alternatives to traditional fossil plastics, 2,5-furandicarboxylic acid (FDCA) represents a potential substitute to terephthalic acid (TPA), a monomer of the ubiquitous polyester, polyethylene terephthalate (PET). <i>Pseudomonas umsongensis</i> GO16, which can metabolise TPA and ethylene glycol (EG), can also oxidise 5-hydroxymethylfurfural (HMF), a precursor to FDCA. The enzymes involved in the oxidation to FDCA, PsfA and PsfG, were identified and characterised. Deletion of FDCA decarboxylase HmfF involved in the conversion of FDCA to furoic acid, and subsequently to a central metabolic intermediate, 2-ketoglutarate, allowed for the accumulation of FDCA. GO16 Δ<i>hmfF</i> cells were grown on glycerol, TPA, EG or mock PET hydrolysate, and the catalyst was then used for the biotransformation of HMF to FDCA. When TPA was used as a growth substrate and to power the biotransformation, the transport of 5-hydroxymethyl-2-furancarboxylic acid (HMFCA) into the cytoplasm represented a rate-limiting step in HMF oxidation. De-bottlenecking transport limitations through <i>in trans</i> overexpression of the HMFCA transporter (HmfT) along with the PsfA aldehyde dehydrogenase and PsfG alcohol dehydrogenase allowed 100% conversion of 50 mM HMF to FDCA within 24 h when TPA, EG or mock PET hydrolysate were used to grow the biocatalyst and subsequently to power the biotransformation. This expands the repertoire of valuable products obtained from engineered <i>P. umsongensis</i> GO16 in the strategy to bio-upcycle post-consumer PET.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 5","pages":""},"PeriodicalIF":5.7000,"publicationDate":"2025-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70159","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Microbial Biotechnology","FirstCategoryId":"5","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1111/1751-7915.70159","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

Abstract

In the search for biobased alternatives to traditional fossil plastics, 2,5-furandicarboxylic acid (FDCA) represents a potential substitute to terephthalic acid (TPA), a monomer of the ubiquitous polyester, polyethylene terephthalate (PET). Pseudomonas umsongensis GO16, which can metabolise TPA and ethylene glycol (EG), can also oxidise 5-hydroxymethylfurfural (HMF), a precursor to FDCA. The enzymes involved in the oxidation to FDCA, PsfA and PsfG, were identified and characterised. Deletion of FDCA decarboxylase HmfF involved in the conversion of FDCA to furoic acid, and subsequently to a central metabolic intermediate, 2-ketoglutarate, allowed for the accumulation of FDCA. GO16 ΔhmfF cells were grown on glycerol, TPA, EG or mock PET hydrolysate, and the catalyst was then used for the biotransformation of HMF to FDCA. When TPA was used as a growth substrate and to power the biotransformation, the transport of 5-hydroxymethyl-2-furancarboxylic acid (HMFCA) into the cytoplasm represented a rate-limiting step in HMF oxidation. De-bottlenecking transport limitations through in trans overexpression of the HMFCA transporter (HmfT) along with the PsfA aldehyde dehydrogenase and PsfG alcohol dehydrogenase allowed 100% conversion of 50 mM HMF to FDCA within 24 h when TPA, EG or mock PET hydrolysate were used to grow the biocatalyst and subsequently to power the biotransformation. This expands the repertoire of valuable products obtained from engineered P. umsongensis GO16 in the strategy to bio-upcycle post-consumer PET.

Abstract Image

合成2,5-呋喃二羧酸假单胞菌GO16中5-羟甲基糠醛代谢的表征与利用
在寻找传统化石塑料的生物基替代品时,2,5-呋喃二甲酸(FDCA)代表了对苯二甲酸(TPA)的潜在替代品,TPA是普遍存在的聚酯,聚对苯二甲酸乙二醇酯(PET)的单体。假单胞菌GO16可以代谢TPA和乙二醇(EG),也可以氧化FDCA的前体5-羟甲基糠醛(HMF)。对参与氧化FDCA、PsfA和PsfG的酶进行了鉴定和表征。FDCA脱羧酶HmfF的缺失涉及FDCA转化为糠酸,随后转化为中心代谢中间体2-酮戊二酸,允许FDCA的积累。GO16 ΔhmfF细胞在甘油、TPA、EG或模拟PET水解物上生长,然后使用催化剂将HMF生物转化为FDCA。当TPA用作生长底物并为生物转化提供动力时,5-羟甲基-2-呋喃羧酸(HMFCA)转运到细胞质中代表了HMF氧化的限速步骤。当使用TPA、EG或模拟PET水解物培养生物催化剂并随后为生物转化提供动力时,通过反式过表达HmfT (HmfT)以及PsfA醛脱氢酶和PsfG醇脱氢酶,可以在24小时内将50 mM HMF 100%转化为FDCA,从而消除运输限制的瓶颈。这扩大了从工程P. umsongensis GO16在消费后PET生物升级循环战略中获得的有价值产品的曲目。
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
Microbial Biotechnology
Microbial Biotechnology Immunology and Microbiology-Applied Microbiology and Biotechnology
CiteScore
11.20
自引率
3.50%
发文量
162
审稿时长
1 months
期刊介绍: Microbial Biotechnology publishes papers of original research reporting significant advances in any aspect of microbial applications, including, but not limited to biotechnologies related to: Green chemistry; Primary metabolites; Food, beverages and supplements; Secondary metabolites and natural products; Pharmaceuticals; Diagnostics; Agriculture; Bioenergy; Biomining, including oil recovery and processing; Bioremediation; Biopolymers, biomaterials; Bionanotechnology; Biosurfactants and bioemulsifiers; Compatible solutes and bioprotectants; Biosensors, monitoring systems, quantitative microbial risk assessment; Technology development; Protein engineering; Functional genomics; Metabolic engineering; Metabolic design; Systems analysis, modelling; Process engineering; Biologically-based analytical methods; Microbially-based strategies in public health; Microbially-based strategies to influence global processes
×
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