Inhibitor tolerance and bioethanol fermentability of levoglucosan-utilizing Escherichia coli were enhanced by overexpression of stress-responsive gene ycfR:The proteomics-guided metabolic engineering

作     者：Dongdong Chang Zia Ul Islam Junfang Zheng Jie Zhao Xiaoyong Cui Zhisheng Yu 

作者机构：College of Resources and EnvironmentUniversity of Chinese Academy of SciencesBeijing100049PR China Department of ChemistryMississippi State UniversityMississippi StateMS 39762USA Department of Biochemistry and Molecular BiologySchool of Basic Medical SciencesCapital Medical UniversityBeijing100069PR China Yanshan Earth Critical Zone and Surface Fluxes Research StationChinese Academy of SciencesNo.380 Huaibei TownHuairou DistrictBeijing101408PR China RCEES-IMCAS-UCAS Joint-Lab of Microbial Technology for Environmental ScienceBeijing100085PR China 

出 版 物：《Synthetic and Systems Biotechnology》 (合成和系统生物技术（英文）)

年 卷 期：2021年第6卷第4期

页      面：384-395页

核心收录：

学科分类：0710[理学-生物学] 071010[理学-生物化学与分子生物学] 081704[工学-应用化学] 07[理学] 08[工学] 0817[工学-化学工程与技术] 

基　　金：The authors gratefully acknowledge the financial support of the National Natural Science Foundation of China(Grants NO.21978287) the Fundamental Research Funds for the Central Universities(Y954035XX2) 

主　　题：Lignocellulosic biomass Levoglucosan Inhibitor Proteomics Gene overexpression Bioethanol 

摘      要：Pretreatment of lignocellulosic biomass is crucial for the release of biofermentable sugars for biofuels production,which could greatly alleviate the burgeoning environment and energy crisis caused by the massive usage of traditional fossil *** is a cost-saving pretreatment process that can readily decompose biomass into levoglucosan,a promising anhydrosugar;however,many undesired toxic compounds inhibitory to downstream microbial fermentation are also generated during the pyrolysis,immensely impeding the bioconversion of levoglucosan-containing ***,we took the first insight into the proteomic responses of a levoglucosan-utilizing and ethanol-producing Escherichia coli to three representative biomass-derived inhibitors,identifying large amounts of differentially expressed proteins(DEPs)that could guide the downstream metabolic engineering for the development of inhibitor-resistant *** up-and eight down-regulated DEPs were further identified as the biomarker stress-responsive proteins candidate for cellular tolerance to multiple *** these biomarker proteins,YcfR exhibiting the highest expression fold-change level was chosen as the target of overexpression to validate proteomics results and develop robust strains with enhanced inhibitor tolerance and fermentation ***,based on four plasmid-borne genes encoding the levoglucosan kinase,pyruvate decarboxylase,alcohol dehydrogenase,and protein YcfR,a new recombinant strain *** LGE-ycfR was successfully created,showing much higher acetic acid-,furfural-,and phenol-tolerance levels compared to the control without overexpression of *** specific growth rate,final cell density,ethanol concentration,ethanol productivity,and levoglucosan consumption rate of the recombinant were also remarkably *** the proteomics-guided metabolic engineering and phenotypic observations,we for the first time corroborated that YcfR is a stress-induced protein responsi