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| 1 | Enhancement of α-ketoisovalerate production by relieving the product inhibition of L-amino acid deaminase from Proteus mirabilis显示文摘L-Amino acid deaminase(LAAD) is a key enzyme in the deamination of L-valine(L-val) to produce α-ketoisovalerate(KIV). However, the product inhibition of LAAD is a major hindrance to industrial KIV production.In the present study, a combination strategy of modification of flexible loop regions around the product binding site and the avoidance of dramatic change of main-chain dynamics was reported to reduce the product inhibition.The four mutant PM-LAAD^(M4)(PM-LAAD^(S98A/T105A/S106A/L341A)) achieved a 6.2-fold higher catalytic efficiency and an almost 6.7-fold reduction in product inhibition than the wild-type enzyme. Docking experiments suggested that weakened interactions between the product and enzyme, and the flexibility of the 'lid' structure relieved LAAD product inhibition. Finally, the whole-cell biocatalyst PM-LAAD^(M4) has been applied to KIV production,the titer and conversion rate of KIV from L-val were 98.5 g·L^-1 and 99.2% at a 3-L scale, respectively. These results demonstrate that the newly engineered catalyst can significantly reduce the product inhibition, that making KIV a prospective product by bioconversion method, and also provide the understanding of the mechanism of the relieved product inhibition of PM-LAAD. | Shanshan Pei Xiaobo Ruan Jia Liu Wei Song Xiulai Chen Qiuling Luo Liming Liu Jing Wu | 2020 | Chinese Journal of Chemical Engineering2020，28，8： | 2 |
| 2 | Improving succinate production by engineering oxygen‑dependent dynamic pathway regulation in Escherichia coli显示文摘Succinate is an important building block for chemical synthesis.However,during the fermentation process,excessive osmotic stress and byproduct accumulation substantively impair the performance of the microbial cell factory.To this end,two strategies were proposed.First,an osmo-tolerant mutant,Escherichia coli FMME-N-2,was screened by combined mutagenesis(ARTP and^(60)Co-γirradiation)to produce 51.8 g L^(−1)succinate with a productivity of 0.81 g L^(−1)h^(−1).Second,an oxygen-dependent bifunctional switch(OBS)was developed with promoter PfnrF8-based activation and tobacco etch virus protease-based inhibition functions.With ribosomal binding site(RBS)and degron optimization of OBS,the optimal strain E.coli FMME-N-30 achieved a succinate titer and productivity of 119 g L^(−1)and 1.65 g L^(−1) h^(−1),respectively,in a 30-L fermentor,while only 7.1 g L^(−1)acetate and no formate or lactate were detected.Compared to the wild-type strain E.coli FMME-N,the succinate titer was increased by 3.3-fold.These results highlight the applicability of OBS for the large-scale production of value-added chemicals. | Cong Gao Wenxiu Tang Liang Guo Guipeng Hu Jia Liu Liming Liu Xiulai Chen | 2022 | Systems Microbiology and Biomanufacturing2022，2，2： | 1 |
| 3 | Efficient synthesis of tyrosol from L-tyrosine via heterologous Ehrlich pathway in Escherichia coli显示文摘For the efficient conversion of L-tyrosine(L-Tyr)to tyrosol,which is an aromatic compound widely used in the pharmaceutical and chemical industries,a novel four-enzyme cascade pathway based on the Ehrlich pathway of Saccharomyces cerevisiae was designed and reconstructed in Escherichia coli.Then,the expression levels of the relevant enzymes were coordinated using a modular approach and gene duplication after the identification of the pyruvate decarboxylase from Candida tropicalis(CtPDC)as the rate-limiting enzymatic step.In situ product removal(ISPR)strategy with XAD4 resins was explored to avoid product inhibition and further improve tyrosol yield.As a result,the titer and conversion rate of tyrosol obtained were 35.7 g·L^(-1) and 93.6%,respectively,in a 3-L bioreactor.Results presented here provide a potential enzymatic process for industrial production of tyrosol from cheap amino acids. | Xiaobo Ruan Sheng Zhang Wei Song Jia Liu Xiulai Chen Liming Liu Jing Wu | 2022 | Chinese Journal of Chemical Engineering2022，35，7： | 1 |
| 4 | Metabolic engineering of Streptomyces to enhance the synthesis of valuable natural products显示文摘The mycelial bacterium Streptomyces is a workhorse for producing natural products,serving as a key source of drugs and other valuable chemicals.However,its complicated life cycle,silent biosynthetic gene clusters(BGCs),and poorly characterized metabolic mechanisms limit efficient production of natural products.There-fore,a metabolic engineering strategy,including traditional and emerging tools from different disciplines,was developed to further enhance natural product synthesis by Streptomyces.Here,current trends in systems metabolic engineering,including tools and strategies,are reviewed.Particularly,this review focuses on recent developments in the selection of methods for regulating the Streptomyces life cycle,strategies for the activation of silent gene clusters,and the exploration of regulatory mechanisms governing antibiotic production.Finally,future challenges and prospects are discussed. | Zuwei Xu Lihao Ji Wenxiu Tang Liang Guo Cong Gao Xiulai Chen Jia Liu Guipeng Hu Liming Liu | 2022 | Engineering Microbiology2022，2，2： | 0 |
| 5 | Advances in microbial engineering for the production of value‑added products in a biorefinery显示文摘Microbial biorefineries to produce chemicals from renewable feedstock provides attractive advantages,including mild reaction conditions and sustainable manufacturing.However,low-efficiency biorefineries always result in an uncompetitive biological process compared to the current petrochemical process.Thus,improving microbial capacity to maximize product yield,productivity,and titer has been recognized as a central goal for bioengineers and biochemists.The knowledge of cellular biochemistry has enabled the regulation of microbial physiology to couple with chemical production.The rapid development in metabolic engineering provides diverse strategies to enhance the efficiency of chemical biosynthesis pathways.New synthetic biology tools as well as novel regulatory targets also offer the opportunity to improve biorefinery environmental adaptivity.In this review,the recent advances in building efficient biorefineries were showcased.In addition,the challenges and future perspectives of microbial host engineering for increased microbial capacity of a biorefinery were discussed. | Cong Gao Liang Guo Wei Song Jing Wu Xiulai Chen Liming Liu | 2023 | Systems Microbiology and Biomanufacturing2023，3，2： | 0 |
| 6 | Current state and future perspectives of cytochrome P450 enzymes for C-H and C=C oxygenation显示文摘Cytochrome P450 enzymes(CYPs)catalyze a series of C-H and C=C oxygenation reactions,including hydroxylation,epoxidation,and ketonization.They are attractive biocatalysts because of their ability to selectively introduce oxygen into inert molecules under mild conditions.This review provides a comprehensive overview of the C-H and C=C oxygenation reactions catalyzed by CYPs and the various strategies for achieving higher selectivity and enzymatic activity.Furthermore,we discuss the application of C-H and C=C oxygenation catalyzed by CYPs to obtain the desired chemicals or pharmaceutical intermediates in practical production.The rapid development of protein engineering for CYPs provides excellent biocatalysts for selective C-H and C=C oxygenation reactions,thereby promoting the development of environmentally friendly and sustainable production processes. | Yu Yan Jing Wu Guipeng Hu Cong Gao Liang Guo Xiulai Chen Liming Liu Wei Song | 2022 | Synthetic and Systems Biotechnology2022，7，3： | 0 |
| 7 | Metabolic engineering strategies for microbial utilization of C1 feedstocks显示文摘The use of abundant and cheap one carbon(C1)feedstocks to produce value-added chemicals is an important approach for achieving carbon neutrality and tackling environmental problems.The conversion of C1 feedstocks to high-value chemicals is dependent on efficient C1 assimilation pathways and microbial chassis adapted for efficient incorporation.Here,we opted to summarize the natural and synthetic C1 assimilation pathways and their key factors for metabolizing C1 feedstock.Accordingly,we discussed the metabolic engineering strategies for enabling the microbial utilization of C1 feedstocks for the bioproduction of value-added chemicals.In addition,we highlighted future perspectives of C1-based biomanufacturing for achieving a low-carbon footprint for the biosynthesis of chemicals. | Jian Zhang Liang Guo Cong Gao Wei Song Jing Wu Liming Liu Xiulai Chen | 2023 | Systems Microbiology and Biomanufacturing2023，3，1： | 0 |
| 8 | Structure demonstration of perovskite oxide and its epitaxial thin films by second harmonic generation显示文摘Perovskite oxide materials and their epitaxial thin films have attracted wide attention in various research fields,due to the new properties discovered at every period.For the epitaxial perovskite oxide thin films,the interaction between the films and substrates could bring in some novel physical properties,such as two-dimensional electron gas[1],positive colossal magnetoresistance[2],polar metal state[3,4],interfacial enhancement effect[5–7].The(dis)appearance and regulation of these properties are usually accompanied with structural distortion changing of films,which may originate from the thickness variation,substrate replacement,or oxygen vacancies[8].Thus,it is crucial to define what structure it is and figure out how the structure evolves. | WANG JieSu GE Chen GUO Er-Jia XU XiuLai WANG Can JIN KuiJuan | 2020 | Science China(Technological Sciences)2020，63，5： | 0 |
| 9 | Metabolic engineering strategies for microbial utilization of methanol显示文摘The increasing shortage of fossil resources and environmental pollution has renewed interest in the synthesis of value-added biochemicals from methanol.However,most of native or synthetic methylotrophs are unable to assimilate methanol at a sufficient rate to produce biochemicals.Thus,the performance of methylotrophs still needs to be optimized to meet the demands of industrial applications.In this review,we provide an in-depth discussion on the properties of natural and synthetic methylotrophs,and summarize the natural and synthetic methanol assimilation pathways.Further,we discuss metabolic engineering strategies for enabling microbial utilization of methanol for the bioproduction of value-added chemicals.Finally,we highlight the potential of microbial engineering for methanol assimilation and offer guidance for achieving a low-carbon footprint for the biosynthesis of chemicals. | Yamei Gan Xin Meng Cong Gao Wei Song Liming Liu Xiulai Chen | 2023 | Engineering Microbiology2023，3，3： | 0 |
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