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enzyme engineering in microbial biosynthesis of terpenoids:progress and perspectives

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生物工程学报 2024年第8期40卷 2473-2488页

作者： Amna Bibi SU Liqiu DAI Zongjie WANG Qinhong Tianjin Institute of Industrial Biotechnology Chinese Academy of SciencesTianjin 300308China National Center of Technology Innovation for Synthetic Biology Tianjin 300308China University of Chinese Academy of Science Beijing 100049China

Terpenoids,known for their structural and functional diversity,are highly valued,especially in food,cosmetics,and cleaning *** biosynthesis has emerged as a sustainable and environmentally friendly approach for the production of ***,the natural enzymes involved in the synthesis of terpenoids have problems such as low activity,poor specificity,and insufficient stability,which limit the biosynthesis *** engineering plays a pivotal role in the microbial synthesis of *** modifying the structures and functions of key enzymes,researchers have significantly improved the catalytic activity,specificity,and stability of enzymes related to terpenoid synthesis,providing strong support for the sustainable production of *** article reviews the strategies for the modification of key enzymes in microbial synthesis of terpenoids,including improving enzyme activity and stability,changing specificity,and promoting mass transfer through multi-enzyme ***,this article looks forward to the challenges and development directions of enzyme engineering in the microbial synthesis of terpenoids.

关键词： terpenoids microbial synthesis enzyme engineering enzyme modification terpene synthases

Application of enzyme engineering in Pharmacy

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Journal of Integrative Medicine（双语） 2020年第2期9卷 5-7页

作者： Qiaole Li Yanhong Li Haoran Niu Biotechnology North China University of Science and TechnologyTangshanHebei063210China

enzyme engineering is an important part of modern *** to its high reaction specificity,high efficiency,mild reaction conditions,and low pollution,it is also an important method widely used in the pharmaceutical *** application of enzymes in medicine is diverse,such as:diagnosis,prevention and treatment of diseases with enzymes,manufacture of various drugs with enzymes,etc.,mainly through manual operations,to obtain enzymes required by the pharmaceutical industry,and through various means enzymes perform their catalytic *** article mainly introduces the application of enzyme engineering in the pharmaceutical field,and also prospects the development trend of enzyme engineering in the pharmaceutical field.

关键词： enzyme engineering Pharmaceutical field enzyme catalysis Application

Book review of“enzyme engineering:Selective catalysts for applications in biotechnology,organic chemistry,and life science”

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Green Synthesis and Catalysis 2023年第2期4卷 77-77页

作者： Manfred T.Reetz Zhoutong Sun Ge Qu Tianjin Institute of Industrial Biotechnology Chinese Academy of SciencesTianjin 300308China Biocatalysis Section Max-Planck-Institut für KohlenforschungKaiser-Wilhelm-Platz 1Mülheiman der Ruhr 45470Germany

This book[1]begins with an introductory chapter in which the history of directed enzyme evolution is briefly presented and the different gene mutagenesis techniques are outlined,followed by a comprehensive chapter des... 详细信息

关键词： Directed evolution Rational design enzyme engineering Methodology Biocatalysis

Harnessing generative AI to decode enzyme catalysis and evolution for enhanced engineering

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National Science Review 2023年第12期10卷 13-25页

作者： Wen Jun Xie Arieh Warshel Department of Medicinal Chemistry Center for Natural Products Drug Discovery and DevelopmentGenetics InstituteUniversity of Florida Department of Chemistry University of Southern California

enzymes, as paramount protein catalysts, occupy a central role in fostering remarkable progress across numerous fields. However, the intricacy of sequence-function relationships continues to obscure our grasp of enzyme behaviors and curtails our capabilities in rational enzyme engineering. Generative artificial intelligence(AI), known for its proficiency in handling intricate data distributions, holds the potential to offer novel perspectives in enzyme research. Generative models could discern elusive patterns within the vast sequence space and uncover new functional enzyme sequences. This review highlights the recent advancements in employing generative AI for enzyme sequence analysis. We delve into the impact of generative AI in predicting mutation effects on enzyme fitness, catalytic activity and stability, rationalizing the laboratory evolution of de novo enzymes, and decoding protein sequence semantics and their application in enzyme engineering. Notably, the prediction of catalytic activity and stability of enzymes using natural protein sequences serves as a vital link, indicating how enzyme catalysis shapes enzyme evolution. Overall,we foresee that the integration of generative AI into enzyme studies will remarkably enhance our knowledge of enzymes and expedite the creation of superior biocatalysts.

关键词： generative AI mutation effects evolution–catalysis relationship enzyme engineering enzyme evolution

Fatty acid hydratase for value-added biotransformation:A review

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Chinese Journal of Chemical engineering 2020年第8期28卷 2051-2063页

作者： Yan Zhang Bekir Engin Eser Peter Kristensen Zheng Guo Department of engineering Faculty of Science and TechnologyAarhus UniversityGustav wieds vej 108000 AarhusDenmark Faculty of engineering and Science Department of Chemistry and BioscienceAalborg University9220 AalborgDenmark

The synthesis of hydroxy fatty acids(HFAs) from renewable oil feedstock by addition of water onto C_C bonds has attracted great attention in recent years. Given that selective asymmetric hydration of non-activated C_C bonds has been proven difficult to achieve with chemical catalysts, enzymatic catalysis by fatty acid hydratases(FAHs) presents an attractive alternative approach to produce value-added HFAs with high regio-, enantioand stereospecificity, as well as excellent atom economy. Even though FAHs have just been investigated as a potential biocatalyst for a decade, remarkable information about FAHs in different aspects is available;however, a comprehensive review has not been archived. Herein, we summarize the research progresses on biochemical characterization, structural and mechanistic determination, enzyme engineering, as well as biotechnological application of FAHs. The current challenges and opportunities for an efficient utilization of FAHs in organic synthesis and industrial applications are critically discussed.

关键词： Hydroxy fatty acids Fatty acid hydratases enzyme characterization Catalytic mechanism enzyme engineering Biotransformation

Engineered microorganisms and enzymes for efficiently synthesizing plant natural products

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Chinese Journal of Chemical engineering 2021年第2期34卷 62-73页

作者： Wenqiang Li Wentao Sun Chun Li Key Lab for Industrial Biocatalysis Ministry of EducationDepartment of Chemical EngineeringTsinghua UniversityBeijing 100084China

Plant natural products are a kind of active substance widely used in pharmaceuticals and ***,the current production mode based on plant culture and extraction suffer complex processes and severe concerns for environmental and *** the increasing awareness of environmental sustainability,engineered microbial cell factories have been an alternative approach to produce natural *** engineering strategies have been utilized in microbial biosynthesis of complex phytochemicals such as dynamic control and substructure ***,enzyme engineering including directed evolution and rational design has been implemented to improve enzyme catalysis efficiency and stability as well as change promiscuity to expand product *** this review,we discussed recent advances in microbial biosynthesis of complex phytochemicals from the following aspects,including pathway construction,strain engineering to boost the production.

关键词： Plant natural products Microorganisms Synthetic biology enzyme engineering

engineering Rubisco to enhance CO_(2) utilization

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Synthetic and Systems Biotechnology 2024年第1期9卷 55-68页

作者： Lei Zhao Zhen Cai Yin Li Yanping Zhang CAS Key Laboratory of Microbial Physiological and Metabolic engineering State Key Laboratory of Microbial ResourcesInstitute of MicrobiologyChinese Academy of SciencesBeijing100101China University of Chinese Academy of Sciences Beijing100049China Seeds Research Syngenta Biotechnology ChinaZhong Guan Cun Life Science ParkBeijingChina

Ribulose-1,5-bisphosphate carboxylase/oxygenase(Rubisco)is a pivotal enzyme that mediates the fixation of CO_(2).As the most abundant protein on earth,Rubisco has a significant impact on global carbon,water,and nitrogen ***,the significantly low carboxylation activity and competing oxygenase activity of Rubisco greatly impede high carbon fixation *** review first summarizes the current efforts in directly or indirectly modifying plant Rubisco,which has been challenging due to its high conservation and limitations in chloroplast transformation ***,recent advancements in understanding Rubisco biogenesis with the assistance of chaperones have enabled successful heterologous expression of all Rubisco forms,including plant Rubisco,in *** breakthrough facilitates the acquisition and evaluation of modified proteins,streamlining the measurement of their ***,the establishment of a screening system in *** opens up possibilities for obtaining high-performance mutant enzymes through directed ***,this review emphasizes the utilization of Rubisco in microorganisms,not only expanding their carbon-fixing capabilities but also holding significant potential for enhancing biotransformation processes.

关键词： Rubisco Plant CO_(2)fixation Microbial Carbon fixation pathway Synthetic biology enzyme engineering

Methanol tolerance upgrading of Proteus mirabilis lipase by machine learning‑assisted directed evolution

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Systems Microbiology and Biomanufacturing 2023年第3期3卷 427-439页

作者： Rui Ma Yingnan Li Meng Zhang Fei Xu Ministry of Education Key Laboratory of Industrial Biotechnology School of BiotechnologyJiangnan UniversityWuxi 214122China

For many crucial industrial applications,enzyme-catalyzed processes take place in harsh organic solvent ***,it remains a challenging problem to improve enzyme stability in organic *** study utilized the MLDE(machine learning-assisted directed evolution)protocol to improve the methanol tolerance of Proteus mirabilis lipase(PML).The machine learning(ML)models were trained based on 266 combinatorial *** top 3 in 22 regression models based on evaluation of tenfold cross-validation,the fitness landscape of the 8000 full-space combinatorial mutants was *** mutants in the restricted library showed higher methanol tolerance,among which the methanol tolerance of G202N/K208G/G266S(NGS)was up to 13-fold compared with the *** dynamics(MD)simulation showed that reconstructing of critical hydrogen bond network in the mutant region of NGS provides a more stable local *** compact structure may improve the methanol tolerance by preventing organic solvent molecules into the activity site and resisting structural *** work provides a successful case of evolution guided by ML for higher organic solvent tolerance of enzyme,and may also be a reference for broad enzyme modifications.

关键词： Organic solvent tolerance enzyme engineering Fitness prediction Site striation mutagenesis Molecular dynamics

Microbial soluble aromatic prenyltransferases for engineered biosynthesis

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Synthetic and Systems Biotechnology 2021年第2期6卷 51-62页

作者： He-Ping Chen Ikuro Abe Graduate School of Pharmaceutical Sciences The University of TokyoBunkyo-kuTokyo113-0033Japan School of Pharmaceutical Sciences South-Central University for NationalitiesWuhanHubei430074PR China Collaborative Research Institute for Innovative Microbiology The University of TokyoYayoi 1-1-1Bunkyo-kuTokyo113-8657Japan

Prenyltransferase(PTase)enzymes play crucial roles in natural product biosynthesis by transferring isoprene unit(s)to target substrates,thereby generating prenylated *** prenylation step leads to a diverse group of natural products with improved membrane affinity and enhanced bioactivity,as compared to the nonprenylated *** last two decades have witnessed increasing studies on the identification,characterization,enzyme engineering,and synthetic biology of microbial PTase family *** herein summarize several examples of microbial soluble aromatic PTases for chemoenzymatic syntheses of unnatural novel prenylated compounds.

关键词： Microbial prenyltransferase Biosynthesis enzyme engineering Prenylation Synthetic biology