Pub Date : 2024-02-17DOI: 10.1016/j.engmic.2024.100141
Ting Cai , Xinyu Gao , Xiaoyan Qi , Xiaolei Wang , Ruijun Liu , Lei Zhang , Xia Wang
The consumption of non-renewable fossil fuels has directly contributed to a dramatic rise in global carbon dioxide (CO2) emissions, posing an ongoing threat to the ecological security of the Earth. Microbial electrosynthesis (MES) is an innovative energy regeneration strategy that offers a gentle and efficient approach to converting CO2 into high-value products. The cathode chamber is a vital component of an MES system and its internal factors play crucial roles in improving the performance of the MES system. Therefore, this review aimed to provide a detailed analysis of the key factors related to the cathode chamber in the MES system. The topics covered include inward extracellular electron transfer pathways, cathode materials, applied cathode potentials, catholyte pH, and reactor configuration. In addition, this review analyzes and discusses the challenges and promising avenues for improving the conversion of CO2 into high-value products via MES.
不可再生化石燃料的消耗直接导致全球二氧化碳(CO2)排放量急剧上升,对地球的生态安全构成持续威胁。微生物电合成(MES)是一种创新的能源再生策略,它提供了一种温和、高效的方法,将二氧化碳转化为高价值产品。阴极室是微生物电合成系统的重要组成部分,其内部因素对提高微生物电合成系统的性能起着至关重要的作用。因此,本综述旨在详细分析与 MES 系统中阴极室有关的关键因素。涉及的主题包括细胞外电子内向转移途径、阴极材料、应用阴极电位、阴极溶液 pH 值和反应器配置。此外,本综述还分析和讨论了通过 MES 将二氧化碳转化为高价值产品所面临的挑战和有前景的途径。
{"title":"Role of the cathode chamber in microbial electrosynthesis: A comprehensive review of key factors","authors":"Ting Cai , Xinyu Gao , Xiaoyan Qi , Xiaolei Wang , Ruijun Liu , Lei Zhang , Xia Wang","doi":"10.1016/j.engmic.2024.100141","DOIUrl":"10.1016/j.engmic.2024.100141","url":null,"abstract":"<div><p>The consumption of non-renewable fossil fuels has directly contributed to a dramatic rise in global carbon dioxide (CO<sub>2</sub>) emissions, posing an ongoing threat to the ecological security of the Earth. Microbial electrosynthesis (MES) is an innovative energy regeneration strategy that offers a gentle and efficient approach to converting CO<sub>2</sub> into high-value products. The cathode chamber is a vital component of an MES system and its internal factors play crucial roles in improving the performance of the MES system. Therefore, this review aimed to provide a detailed analysis of the key factors related to the cathode chamber in the MES system. The topics covered include inward extracellular electron transfer pathways, cathode materials, applied cathode potentials, catholyte pH, and reactor configuration. In addition, this review analyzes and discusses the challenges and promising avenues for improving the conversion of CO<sub>2</sub> into high-value products via MES.</p></div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"4 3","pages":"Article 100141"},"PeriodicalIF":0.0,"publicationDate":"2024-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2667370324000043/pdfft?md5=0045968362299ca70bded635c93f6f6d&pid=1-s2.0-S2667370324000043-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139966064","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Adenoviruses typically cause mild illnesses, but severe diseases may occur primarily in immunodeficient individuals, particularly children. Recently, adenoviruses have garnered significant interest as a versatile tool in gene therapy, tumor treatment, and vaccine vector development. Over the past two decades, the advent of recombineering, a method based on homologous recombination, has notably enhanced the utility of adenoviral vectors in therapeutic applications. This review summarizes recent advancements in the use of human adenoviral vectors in medicine and discusses the pivotal role of recombineering in the development of these vectors. Additionally, it highlights the current achievements and potential future impact of therapeutic adenoviral vectors.
{"title":"Role of homologous recombination/recombineering on human adenovirus genome engineering: Not the only but the most competent solution","authors":"Lisa-Marie Dawson , Montaha Alshawabkeh , Katrin Schröer , Fatima Arakrak, Anja Ehrhardt, Wenli Zhang","doi":"10.1016/j.engmic.2024.100140","DOIUrl":"10.1016/j.engmic.2024.100140","url":null,"abstract":"<div><p>Adenoviruses typically cause mild illnesses, but severe diseases may occur primarily in immunodeficient individuals, particularly children. Recently, adenoviruses have garnered significant interest as a versatile tool in gene therapy, tumor treatment, and vaccine vector development. Over the past two decades, the advent of recombineering, a method based on homologous recombination, has notably enhanced the utility of adenoviral vectors in therapeutic applications. This review summarizes recent advancements in the use of human adenoviral vectors in medicine and discusses the pivotal role of recombineering in the development of these vectors. Additionally, it highlights the current achievements and potential future impact of therapeutic adenoviral vectors.</p></div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"4 1","pages":"Article 100140"},"PeriodicalIF":0.0,"publicationDate":"2024-02-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2667370324000031/pdfft?md5=06fac970e5d6d24bc3d15d640a5837dc&pid=1-s2.0-S2667370324000031-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139827339","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-02-02DOI: 10.1016/j.engmic.2024.100139
Zhongye Li , Pankajkumar R. Waghmare , Lubbert Dijkhuizen , Xiangfeng Meng , Weifeng Liu
Lignocellulosic biomass is an abundant and renewable bioresource for the production of biofuels and biochemical products. The classical biorefinery process for lignocellulosic degradation and conversion comprises three stages, i.e., pretreatment, enzymatic saccharification, and fermentation. However, the complicated pretreatment process, high cost of cellulase production, and insufficient production performance of fermentation strains have restricted the industrialization of biorefinery. Consolidated bioprocessing (CBP) technology combines the process of enzyme production, enzymatic saccharification, and fermentation in a single bioreactor using a specific microorganism or a consortium of microbes and represents another approach worth exploring for the production of chemicals from lignocellulosic biomass. The present review summarizes the progress made in research of CBP technology for lignocellulosic biomass conversion. In this review, different CBP strategies in lignocellulose biorefinery are reviewed, including CBP with natural lignocellulose-degrading microorganisms as the chassis, CBP with biosynthetic microorganisms as the chassis, and CBP with microbial co-culturing systems. This review provides new perspectives and insights on the utilization of low-cost feedstock lignocellulosic biomass for production of biochemicals.
{"title":"Research advances on the consolidated bioprocessing of lignocellulosic biomass","authors":"Zhongye Li , Pankajkumar R. Waghmare , Lubbert Dijkhuizen , Xiangfeng Meng , Weifeng Liu","doi":"10.1016/j.engmic.2024.100139","DOIUrl":"10.1016/j.engmic.2024.100139","url":null,"abstract":"<div><p>Lignocellulosic biomass is an abundant and renewable bioresource for the production of biofuels and biochemical products. The classical biorefinery process for lignocellulosic degradation and conversion comprises three stages, i.e., pretreatment, enzymatic saccharification, and fermentation. However, the complicated pretreatment process, high cost of cellulase production, and insufficient production performance of fermentation strains have restricted the industrialization of biorefinery. Consolidated bioprocessing (CBP) technology combines the process of enzyme production, enzymatic saccharification, and fermentation in a single bioreactor using a specific microorganism or a consortium of microbes and represents another approach worth exploring for the production of chemicals from lignocellulosic biomass. The present review summarizes the progress made in research of CBP technology for lignocellulosic biomass conversion. In this review, different CBP strategies in lignocellulose biorefinery are reviewed, including CBP with natural lignocellulose-degrading microorganisms as the chassis, CBP with biosynthetic microorganisms as the chassis, and CBP with microbial co-culturing systems. This review provides new perspectives and insights on the utilization of low-cost feedstock lignocellulosic biomass for production of biochemicals.</p></div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"4 2","pages":"Article 100139"},"PeriodicalIF":0.0,"publicationDate":"2024-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S266737032400002X/pdfft?md5=c68277253e9d4a1532c115f4aa451087&pid=1-s2.0-S266737032400002X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139883128","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-18DOI: 10.1016/j.engmic.2024.100138
Zhouhai Zhu , Meng Wang , Ying Guan , Meng Li , Qiyuan Peng , Ning Zheng , Wenbin Ma
The association between cigarette smoking and the gut microbiota remains unclear, and there is no agreement on how smoking affects the composition of gut microorganisms. In this study, the relationship between smoking status and gut microbial composition was investigated by performing 16S rRNA gene amplicon sequencing analysis of stool samples from 80 healthy Chinese adults. The results showed that smoking did not cause significant changes to the composition and microbial functional pathways of the gut microbiota. However, smoking altered the relative abundance of several specific taxa, where Phascolarctobacterium and Fusobacterium increased and Dialister decreased. Notably, our analysis revealed that smoking introduced more microbial interactions to the interaction network and decreased its modularity. Overall, this study provides new insights into the association between smoking status and the gut microbiota.
{"title":"Altered interaction network in the gut microbiota of current cigarette smokers","authors":"Zhouhai Zhu , Meng Wang , Ying Guan , Meng Li , Qiyuan Peng , Ning Zheng , Wenbin Ma","doi":"10.1016/j.engmic.2024.100138","DOIUrl":"10.1016/j.engmic.2024.100138","url":null,"abstract":"<div><p>The association between cigarette smoking and the gut microbiota remains unclear, and there is no agreement on how smoking affects the composition of gut microorganisms. In this study, the relationship between smoking status and gut microbial composition was investigated by performing 16S rRNA gene amplicon sequencing analysis of stool samples from 80 healthy Chinese adults. The results showed that smoking did not cause significant changes to the composition and microbial functional pathways of the gut microbiota. However, smoking altered the relative abundance of several specific taxa, where <em>Phascolarctobacterium</em> and <em>Fusobacterium</em> increased and <em>Dialister</em> decreased. Notably, our analysis revealed that smoking introduced more microbial interactions to the interaction network and decreased its modularity. Overall, this study provides new insights into the association between smoking status and the gut microbiota.</p></div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"4 2","pages":"Article 100138"},"PeriodicalIF":0.0,"publicationDate":"2024-01-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2667370324000018/pdfft?md5=8daccfb83c08c8c63fcdd652a9093808&pid=1-s2.0-S2667370324000018-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139637241","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-12-19DOI: 10.1016/j.engmic.2023.100137
Shuang Zhao , Ruiying Feng , Yuan Gu , Liyuan Han , Xiaomei Cong , Yang Liu , Shuo Liu , Qiyao Shen , Liujie Huo , Fu Yan
Microbial natural products and their derivatives have been developed as a considerable part of clinical drugs and agricultural chemicals. Marine microbial natural products exhibit diverse chemical structures and bioactivities with substantial potential for the development of novel pharmaceuticals. However, discovering compounds with new skeletons from marine microbes remains challenging. In recent decades, multiple approaches have been developed to discover novel marine microbial natural products, among which heterologous expression has proven to be an effective method. Facilitated by large DNA cloning and comparative metabolomic technologies, a few novel bioactive natural products from marine microorganisms have been identified by the expression of their biosynthetic gene clusters (BGCs) in heterologous hosts. Heterologous expression is advantageous for characterizing gene functions and elucidating the biosynthetic mechanisms of natural products. This review provides an overview of recent progress in heterologous expression-guided discovery, biosynthetic mechanism elucidation, and yield optimization of natural products from marine microorganisms and discusses the future directions of the heterologous expression strategy in facilitating novel natural product exploitation.
微生物天然产品及其衍生物已被开发为临床药物和农用化学品的重要组成部分。海洋微生物天然产物表现出多种化学结构和生物活性,具有开发新型药物的巨大潜力。然而,从海洋微生物中发现具有新骨架的化合物仍然具有挑战性。近几十年来,人们开发了多种方法来发现新型海洋微生物天然产物,其中异源表达被证明是一种有效的方法。在大 DNA 克隆和比较代谢组学技术的推动下,通过在异源宿主中表达海洋微生物的生物合成基因簇(BGCs),从海洋微生物中发现了一些新型生物活性天然产物。异源表达有利于鉴定基因功能和阐明天然产物的生物合成机制。本综述概述了在异源表达引导下发现、阐明生物合成机制以及优化海洋微生物天然产物产量方面的最新进展,并探讨了异源表达策略在促进新型天然产物开发方面的未来发展方向。
{"title":"Heterologous expression facilitates the discovery and characterization of marine microbial natural products","authors":"Shuang Zhao , Ruiying Feng , Yuan Gu , Liyuan Han , Xiaomei Cong , Yang Liu , Shuo Liu , Qiyao Shen , Liujie Huo , Fu Yan","doi":"10.1016/j.engmic.2023.100137","DOIUrl":"10.1016/j.engmic.2023.100137","url":null,"abstract":"<div><p>Microbial natural products and their derivatives have been developed as a considerable part of clinical drugs and agricultural chemicals. Marine microbial natural products exhibit diverse chemical structures and bioactivities with substantial potential for the development of novel pharmaceuticals. However, discovering compounds with new skeletons from marine microbes remains challenging. In recent decades, multiple approaches have been developed to discover novel marine microbial natural products, among which heterologous expression has proven to be an effective method. Facilitated by large DNA cloning and comparative metabolomic technologies, a few novel bioactive natural products from marine microorganisms have been identified by the expression of their biosynthetic gene clusters (BGCs) in heterologous hosts. Heterologous expression is advantageous for characterizing gene functions and elucidating the biosynthetic mechanisms of natural products. This review provides an overview of recent progress in heterologous expression-guided discovery, biosynthetic mechanism elucidation, and yield optimization of natural products from marine microorganisms and discusses the future directions of the heterologous expression strategy in facilitating novel natural product exploitation.</p></div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"4 2","pages":"Article 100137"},"PeriodicalIF":0.0,"publicationDate":"2023-12-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2667370323000693/pdfft?md5=327efcf30168d96356c4e7af90784416&pid=1-s2.0-S2667370323000693-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138991756","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Peptaibiotics are linear or cyclic peptide antibiotics characterized by the non-proteinogenic amino acid, alpha-aminoisobutyric acid. They exhibit a wide range of bioactivity against various pathogens. This report presents a comprehensive review of analytical methods for Trichoderma cultivation, production, isolation, screening, purification, and characterization of peptaibiotics, along with their bioactivity. Numerous techniques are currently available for each step, and we focus on describing the most commonly used and recently developed chromatographic and spectroscopic techniques. Investigating peptaibiotics requires efficient culture media, growth conditions, and isolation and purification techniques. The combination of chromatographic and spectroscopic tools offers a better opportunity for characterizing and identifying peptaibiotics. The evaluation of the chemical and biological properties of this compound has also been explored concerning its potential application in pharmaceutical and other industries. This review aims to summarize available data on the techniques and tools used to screen and purify peptaibiotics from Trichoderma fungi and bioactivity against various pathogens.
{"title":"Chromatographic and Mass Spectroscopic Guided Discovery of Trichoderma Peptaibiotics and their Bioactivity","authors":"Adigo Setargie , Chen Wang , Liwen Zhang , Yuquan Xu","doi":"10.1016/j.engmic.2023.100135","DOIUrl":"10.1016/j.engmic.2023.100135","url":null,"abstract":"<div><p>Peptaibiotics are linear or cyclic peptide antibiotics characterized by the non-proteinogenic amino acid, alpha-aminoisobutyric acid. They exhibit a wide range of bioactivity against various pathogens. This report presents a comprehensive review of analytical methods for <em>Trichoderma</em> cultivation, production, isolation, screening, purification, and characterization of peptaibiotics, along with their bioactivity. Numerous techniques are currently available for each step, and we focus on describing the most commonly used and recently developed chromatographic and spectroscopic techniques. Investigating peptaibiotics requires efficient culture media, growth conditions, and isolation and purification techniques. The combination of chromatographic and spectroscopic tools offers a better opportunity for characterizing and identifying peptaibiotics. The evaluation of the chemical and biological properties of this compound has also been explored concerning its potential application in pharmaceutical and other industries. This review aims to summarize available data on the techniques and tools used to screen and purify peptaibiotics from <em>Trichoderma</em> fungi and bioactivity against various pathogens.</p></div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"4 2","pages":"Article 100135"},"PeriodicalIF":0.0,"publicationDate":"2023-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S266737032300067X/pdfft?md5=ae89c70326f4df088d4fe03f65e7a184&pid=1-s2.0-S266737032300067X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139026613","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-12-15DOI: 10.1016/j.engmic.2023.100136
Xueshen Wu , Chao Wang , Depeng Wang , Ahmed Tawfik , Ronghua Xu , Zhong Yu , Fangang Meng
In this study, a combined system consisting of an anaerobic membrane bioreactor (AnMBR) and flow-through biofilm reactor/CANON (FTBR/CANON) was developed to simultaneously remove carbon and nitrogen from synthetic livestock wastewater. The average removal efficiencies of total nitrogen (TN) were 64.2 and 76.4% with influent ammonium (NH4+-N) concentrations of approximately 200 and 500 mg/L, respectively. The COD removal efficiencies were higher than 98.0% during the entire operation. Mass balance analysis showed that COD and TN were mainly removed by the AnMBR and FTBR/CANON, respectively. The anammox process was the main nitrogen removal pathway in the combined system, with a contribution of over 80%. High functional bacterial activity was observed in the combined system. Particularly, an increase in the NH4+-N concentration considerably improved the anammox activity of the biofilm in the FTBR/CANON. 16S rRNA high-throughput sequencing revealed that Methanosaeta, Candidatus Methanofastidiosum, and Methanobacterium were the dominant methanogens in the AnMBR granular sludge. In the CANON biofilm, Nitrosomonas and Candidatus Kuenenia were identified as aerobic and anaerobic ammonium-oxidizing bacteria, respectively. In summary, this study proposes a combined AnMBR and FTBR/CANON process targeting COD and nitrogen removal, and provides a potential alternative for treating high-strength wastewater.
{"title":"Achieving simultaneous removal of carbon and nitrogen by an integrated process of anaerobic membrane bioreactor and flow-through biofilm reactor","authors":"Xueshen Wu , Chao Wang , Depeng Wang , Ahmed Tawfik , Ronghua Xu , Zhong Yu , Fangang Meng","doi":"10.1016/j.engmic.2023.100136","DOIUrl":"10.1016/j.engmic.2023.100136","url":null,"abstract":"<div><p>In this study, a combined system consisting of an anaerobic membrane bioreactor (AnMBR) and flow-through biofilm reactor/CANON (FTBR/CANON) was developed to simultaneously remove carbon and nitrogen from synthetic livestock wastewater. The average removal efficiencies of total nitrogen (TN) were 64.2 and 76.4% with influent ammonium (NH<sub>4</sub><sup>+</sup>-N) concentrations of approximately 200 and 500 mg/L, respectively. The COD removal efficiencies were higher than 98.0% during the entire operation. Mass balance analysis showed that COD and TN were mainly removed by the AnMBR and FTBR/CANON, respectively. The anammox process was the main nitrogen removal pathway in the combined system, with a contribution of over 80%. High functional bacterial activity was observed in the combined system. Particularly, an increase in the NH<sub>4</sub><sup>+</sup>-N concentration considerably improved the anammox activity of the biofilm in the FTBR/CANON. 16S rRNA high-throughput sequencing revealed that <em>Methanosaeta, Candidatus Methanofastidiosum</em>, and <em>Methanobacterium</em> were the dominant methanogens in the AnMBR granular sludge. In the CANON biofilm, <em>Nitrosomonas</em> and <em>Candidatus</em> Kuenenia were identified as aerobic and anaerobic ammonium-oxidizing bacteria, respectively. In summary, this study proposes a combined AnMBR and FTBR/CANON process targeting COD and nitrogen removal, and provides a potential alternative for treating high-strength wastewater.</p></div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"4 1","pages":"Article 100136"},"PeriodicalIF":0.0,"publicationDate":"2023-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2667370323000681/pdfft?md5=8afe17500f65f7f5888c34f6b953307b&pid=1-s2.0-S2667370323000681-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139018324","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-11-30DOI: 10.1016/j.engmic.2023.100134
Jiangyu Yang , Kun Li , Yongheng Rong , Zhaoxi Liu , Xiaoyu Liu , Yue Yu , Wenjing Shi , Yun Kong , Min Chen
The modularity of carbohydrate-active enzymes facilitates that enzymes with different functions have similar fragments. However, because of the complex structure of the enzyme active sites and the epistatic effects of various mutations on enzyme activity, it is difficult to design enzymes with multiple mutation sites using conventional methods. In this study, we designed multi-point mutants by fragment replacement in the donor-acceptor binding pocket of Actinobacillus pleuropneumoniae N-glycosyltransferase (ApNGT) to obtain novel properties. Candidate fragments were selected from a customized glycosyltransferase database. The stability and substrate-binding energy of the three fragment replacement mutants were calculated in comparison with wild-type ApNGT, and mutants with top-ranking stability and middle-ranking substrate-binding energy were chosen for priority experimental verification. We found that a mutant called F13, which increased the glycosylation efficiency of the natural substrate by 1.44 times, the relative conversion of UDP-galactose by 14.2 times, and the relative conversion of UDP-xylose from almost 0 to 78.6%. Most importantly, F13 mutant acquired an entirely new property, the ability to utilize UDP-glucuronic acid. On one hand, this work shows that replacing similar fragments in the donor-acceptor binding pocket of the enzyme might provide new ideas for designing mutants with new properties; on the other hand, F13 mutant is expected to play an important role in targeted drug delivery.
{"title":"Rational design of a highly active N-glycosyltransferase mutant using fragment replacement approach","authors":"Jiangyu Yang , Kun Li , Yongheng Rong , Zhaoxi Liu , Xiaoyu Liu , Yue Yu , Wenjing Shi , Yun Kong , Min Chen","doi":"10.1016/j.engmic.2023.100134","DOIUrl":"10.1016/j.engmic.2023.100134","url":null,"abstract":"<div><p>The modularity of carbohydrate-active enzymes facilitates that enzymes with different functions have similar fragments. However, because of the complex structure of the enzyme active sites and the epistatic effects of various mutations on enzyme activity, it is difficult to design enzymes with multiple mutation sites using conventional methods. In this study, we designed multi-point mutants by fragment replacement in the donor-acceptor binding pocket of <em>Actinobacillus pleuropneumoniae N</em>-glycosyltransferase (ApNGT) to obtain novel properties. Candidate fragments were selected from a customized glycosyltransferase database. The stability and substrate-binding energy of the three fragment replacement mutants were calculated in comparison with wild-type ApNGT, and mutants with top-ranking stability and middle-ranking substrate-binding energy were chosen for priority experimental verification. We found that a mutant called F13, which increased the glycosylation efficiency of the natural substrate by 1.44 times, the relative conversion of UDP-galactose by 14.2 times, and the relative conversion of UDP-xylose from almost 0 to 78.6%. Most importantly, F13 mutant acquired an entirely new property, the ability to utilize UDP-glucuronic acid. On one hand, this work shows that replacing similar fragments in the donor-acceptor binding pocket of the enzyme might provide new ideas for designing mutants with new properties; on the other hand, F13 mutant is expected to play an important role in targeted drug delivery.</p></div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"4 1","pages":"Article 100134"},"PeriodicalIF":0.0,"publicationDate":"2023-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2667370323000668/pdfft?md5=9d882243453dbe21a97fd045b480ba38&pid=1-s2.0-S2667370323000668-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139297743","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}