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Emulsion‑based evolution of Escherichia coli for higher growth yield on D‑xylose identifies central role of cyclic AMP

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Systems Microbiology and Biomanufacturing 2023年第4期3卷 730-738页

作者： James S.Orr Edwin Zen Xiaoyi Wang Christopher V.Rao DOE Center for Advanced Bioenergy and Bioproducts Innovation UrbanaILUSA Department of Chemical and Biomolecular Engineering University of Illinois at Urbana-Champaign600 S.Mathews Ave.UrbanaIL 61801USA Core Facility Center The First Affiliated Hospital of Nanjing Medical UniversityNanjingChina

D-xylose is an abundant sugar found in plant biomass and can be used as a renewable feedstock for the microbial production of diverse biofuels and ***,D-xylose metabolism is slow in many industrial microorganisms,at least as compared to glucose *** surprisingly,a number of approaches have been developed for improving D-xylose metabolism in diverse *** this work,we applied a previously developed evolution strategy based on media-in-oil emulsions for improving the growth yield of Escherichia coli NCM3722 on *** 30 rounds of evolutions,we isolated multiple mutants with increased growth yield on *** addition,we also observed similar increases in the growth *** mutants were selected for whole-genome *** mutants had an amber stop mutation in adenylate cyclase,which truncates nearly 60%of the ***,the ability of this mutant to grow on xylose indicated that truncated enzyme,lacking the C-terminal regulatory domain,is still *** other mutant had a point mutation in the cyclic AMP receptor protein(CRP),near the high affinity binding site for cyclic *** mutations,when intro-duced into wild type ***,were able to increase the growth yields at levels similar to the isolated *** addition to D-xylose,these mutant strains and their genetic mimics also exhibited higher growth rates and yields on glucose,lactose,and *** results suggest that the improved growth rates and yields are due to changes in the production and sensing of intracellular cyclic AMP concentrations and also suggest native concentrations are suboptimal with respect to the growth rate and yield under the growth conditions ***,these results may prove useful for engineering strains of *** for high-density fermentations or protein production.

关键词： Escherichia coli Xylose cAMP adaptive laboratory evolution

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Formate as a supplementary substrate facilitates sugar metabolism and solvent production by Clostridium beijerinckii NCIMB 8052

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Synthetic and Systems Biotechnology 2023年第2期8卷 196-205页

作者： Ran Zhao Wenyue Dong Chen Yang Weihong Jiang Jinzhong Tian Yang Gu CAS-Key laboratory of Synthetic Biology CAS Center for Excellence in Molecular Plant SciencesInstitute of Plant Physiology and EcologyChinese Academy of SciencesShanghai200032China University of Chinese Academy of Sciences Beijing100049China

Microbial utilization and conversion of organic one-carbon compounds,such as formate and methanol that can be easily produced from CO_(2),has emerged as an attractive approach for *** this study,we discovered Clostridium beijerinckii NCIMB 8052,a typical solventogenic Clostridium strain,to be a native formate-utilizing bacterium.^(13)C isotope analysis showed that formate could be metabolized via both assimilation and dissimi-lation pathways in *** NCIMB ***,the use of formate as the supplementary substrate by this strain could significantly enhance its glucose consumption and ABE(acetone-butanol-ethanol)production,largely due to the up-regulation of genes responsible for glycolysis and glucose transport under formate *** on these findings,we further improved formate tolerance of *** NCIMB 8052 by adaptive lab-oratory evolution,generating an evolved strain *** Cbei-FA01 strain could produce 23.0 g/L of ABE solvents using glucose and formate as dual substrates,~50%higher than that of the wild-type strain under the same ***,such a promotion effect of formate on ABE production by Cbei-FA01 was also observed in fermenting a glucose-xylose *** work reveals a previously unreported role of formate in biological ABE production,providing a new approach to utilize this one-carbon source.

关键词： Formate utilization Clostridium beijerinckii Metabolic pathways adaptive laboratory evolution ABE solvents

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Enhancing the light reactions of photosynthesis:Strategies,controversies,and perspectives

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Molecular Plant 2023年第1期16卷 4-22页

作者： Dario Leister Plant Molecular Biology Faculty of BiologyLudwig-Maximilians-University(LMU)MunichMartinsried-PlaneggD-82152 MunichGermany

Photosynthesis is central to life on Earth,employing sunlight,water,and carbon dioxide to produce chemical energy and *** is generally accepted that boosting its efficiency offers one promising way to increase crop yields under agronomically realistic *** the components,structure,and regulatory mechanisms of the light reactions of photosynthesis are well understood,concepts for enhancing the process have been suggested and partially *** approaches vary in complexity,from targeting single components to comprehensive redesign of the whole process on the scales from single cells or tissues to whole *** to enhance light utilization per leaf,by decreasing pigmentation,increasing levels of photosynthetic proteins,prolonging the lifespan of the photosynthetic machinery,or massive reconfiguration of the photosynthetic machinery and the incorporation of nanomaterials,are discussed in this review ***,strategies intended to optimize the acclimation of photosynthesis to changes in the environment are presented,including redesigning mechanisms to dissipate excess excitation energy(e.g.,non-photochemical quenching)or reduction power(e.g.,flavodiiron proteins).Moreover,schemes for improving acclimation,inspired by natural or laboratory-induced adaptation,are ***,all these endeavors are still in an early exploratory phase and/or have not resulted in the desired outcome,largely because photosynthesis is embedded within large networks of closely interacting cellular and metabolic processes,which can vary among species and even *** explains why integrated,systems-wide approaches are required to achieve the breakthroughs required for effectively increasing crop yields.

关键词： Arabidopsis adaptive laboratory evolution crop yield photosynthesis synthetic biology tobacco

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From nature to nurture:Essence and methods to isolate robust methanotrophic bacteria

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Synthetic and Systems Biotechnology 2020年第3期5卷 173-178页

作者： Haritha Meruvu Hui Wu Ziyue Jiao Liyan Wang Qiang Fei School of Chemical Engineering and Technology Xi'an Jiaotong UniversityXi'anShaanxiChina State Key laboratory of Bioreactor Engineering East China University of Science and TechnologyShanghaiChina Luoyang TMAXTREE Biotechnology Co. Ltd.LuoyangChina Shaanxi Key laboratory of Energy Chemical Process Intensification Xi'an Jiaotong UniversityXi'anChina

Methanotrophic bacteria are entities with innate biocatalytic potential to biofilter and oxidize methane into simpler compounds concomitantly conserving energy,which can contribute to copious industrial *** future and efficacy of such industrial applications relies upon acquiring and/or securing robust methanotrophs with taxonomic and phenotypic *** several dramatic advances,isolation of robust methanotrophs is still a long-way challenging task with several lacunae to be filled in *** with high tolerance to methane can be isolated and cultivated by mimicking natural environs,and adopting strategies like adaptive metabolic *** review summarizes existent and innovative methods for methanotrophic isolation and purification,and their respective applications.A comprehensive description of new insights shedding light upon how to isolate and concomitantly augment robust methanotrophic metabolism in an orchestrated fashion follows.

关键词： Methanotrophs Isolation Culture medium High throughput adaptive laboratory evolution

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ARTP-ALE Optimized Selection of Low Acetaldehyde Producing Brewer's Yeast and Fermentation Validation

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Journal of Northeast Agricultural University(English Edition) 2023年第2期30卷 8-19页

作者： Chen Zuo-hui Dong Chun-xiu Sun Yao Hui Mi-zhou Zhang Hui Tang Fei Shuang Bao College of Agriculture Northeast Agricultural UniversityHarbin 150030China

Higher levels of acetaldehyde in beer are one of the major concerns in the current beer *** produces acetaldehyde during alcoholic fermentation,and its modification significantly affects beer flavor and quality.A different mutant strain with lower acetaldehyde production and improved ethanol tolerance was constructed using the ARTP-ALE mutagenesis strategy with *** a result of the mutation,the alcohol dehydrogenase activity of the mutant strain decreased to about 71.22%of that of the wild-type *** the same time,the fermentation properties and genetic stability of the newly screened strain showed slight differences from the wild-type strain,and there were no safety concerns regarding industrial use of the mutant strain.

关键词： brewer's yeast acetaldehyde atmospheric pressure room temperature plasma adaptive laboratory evolution ethanol dehydrogenase acetaldehyde dehydrogenase

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Investigating formate tolerance mechanisms in Saccharomyces cerevisiae and its application

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Green Carbon 2023年第1期1卷 65-74页

作者： Kai Wang Yining Liu Zhuoheng Wu Yilu Wu Haoran Bi Yanhui Liu Meng Wang Biqiang Chen Jens Nielsen Zihe Liu Tianwei Tan National Energy R&D Center for Biorefnery Beijing Key Lab of BioprocessCollege of Life Science and TechnologyBeijing Advanced Innovation Center for Soft MatterScience and EngineeringBeijing University of Chemical TechnologyNo.15 North 3rd Ring Rd East100029 BeijingPR China Department of Biology and Biological Engineering Chalmers University of TechnologySE41296 GothenburgSweden

Current global energy and environmental crisis have spurred efforts towards developing sustainable biotechnological solutions,such as utilizing CO_(2) and its derivatives as raw *** is an attractive onecarbon source due to its high solubility and low reduction ***,the regulatory mechanism of formate metabolism in yeast remains largely *** study employed adaptive laboratory evolution(ALE)to improve formate tolerance in Saccharomyces cerevisiae and characterized the underlying molecular *** evolved strain was applied to produce free fatty acids(FFAs)under high concentration of formate with glucose *** results showed that the evolved strain achieved a FFAs titer of 250 mg/***,this study sheds light on the regulatory mechanism of formate tolerance and provides a platform for future studies under high concentrations of formate.

关键词： Formate tolerance adaptive laboratory evolution Saccharomyces cerevisiae Free fatty acids

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