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Coupling biomass pretreatment for enzymatic hydrolysis and direct biomass-to-electricity conversion with molybdovanadophosphoric heteropolyacids as anode electron transfer carriers

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Journal of Energy Chemistry 2021年第7期30卷 133-146页

作者： Huishan Yang Yuchen Bai Denghao Ouyang Fangqian Wang Dehua Liu Xuebing Zhao Key laboratory of Industrial biocatalysis Ministry of EducationTsinghua UniversityBeijing 100084China Institute of applied Chemistry Department of Chemical EngineeringTsinghua UniversityBeijing 100084China

Owing to their acidity,oxidizing ability and redox reversibility,molybdovanadophosphoric heteropolyacids(H_(n+3)PMo_(12-n)VnO40,abbreviated as PMo_(12-n)Vn) were employed as electron transfer carriers for coupling biomass pretreatment for enzymatic hydrolysis and direct biomass-to-electricity *** this novel coupled process,PMo_(12-n)Vn pretreatment that causes deconstruction of cell wall structure with PMo_(12-n)Vn being simultaneously reduced can be considered as the "charging" *** reduced PMo_(12-n)Vn are further re-oxidized with release of electrons in a liquid flow fuel cell(LFFC) to generate electricity is the "discharging" *** Keggin-type PMo_(12-n)Vn with different degree of vanadium substitution(DSV, namely n) were *** to Keggin-type phosphomolybdic acid(PMo_(12)),PMo_(12-n)Vn(n=1-6) showed higher oxidizing ability but poorer redox *** cellulose enzymatic digestibility of PMo_(12-n)Vn pretreated wheat straw generally decreased with increase in DSV, but xylan enzymatic digestibility generally increased with ***_(12) pretreatment of wheat straw at 120℃ obtained the highest enzymatic glucan conversion(EGC) reaching 95%,followed by PMo11V1 pretreatment(85%).Discharging of the reduced heteropolyacids in LFFC showed that vanadium substitution could improve the maximum output power density(Pmax).The highest Pmax was obtained by PMo9 V3(44.7 mW/cm^(2)) when FeCl_(3) was used as a cathode electron carrier,while PMo_(12) achieved the lowest Pmax(27.4 mW/cm^(2)).All the heteropolyacids showed good electrode Faraday efficiency(>95%) and cell discharging efficiency(>93%).The energy efficiency of the coupled process based on the heat values of the products and generated electric energy was in the range of 18%-25% depending on ***_(12) and PMo11V1 seem to be the most suitable heteropolyacids to mediate the coupled process.

关键词： Molybdovanadophosphoric heteropolyacid Electron transfer carrier Biomass pretreatment Enzymatic hydrolysis Direct biomass fuel cell Electricity generation

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Electrochemical reduction of carbon dioxide to produce formic acid coupled with oxidative conversion of biomass

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Journal of Energy Chemistry 2024年第5期92卷 705-729页

作者： Xi Liu Yifan Wang Zhiwei Dai Daihong Gao Xuebing Zhao Key laboratory of Industrial biocatalysis Ministry of EducationTsinghua UniversityBeijing 100084China Institute of applied Chemistry Department of Chemical EngineeringTsinghua UniversityBeijing 100084China

Electrochemical reduction of CO_(2)(CO_(2)RR)has become a research hot spot in recent years in the context of carbon *** is one of the most promising products obtained by electrochemical reduction of CO_(2) due to its high energy value as estimated by market price per energy unit and wide application in chemical *** is the most abundant renewable resource in the natural *** biomass oxidative conversion with CO_(2)RR driven by renewable electricity would well achieve carbon *** this work,we comprehensively reviewed the current research progress on CO_(2)RR to produce HCOOH and coupled system for conversion of biomass and its derivatives to produce value-added ***-and Bi-based electrocatalysts are discussed for CO_(2)RR with regards to the structure of the catalyst and reaction ***-oxidation reactions of biomass derived sugars,alcohols,furan aldehydes and even polymeric components of lignocellulose were reviewed as alternatives to replace oxygen evolution reaction(OER)in the conventional electrolysis *** was recommended that to further improve the efficiency of the coupled system,future work should be focused on the development of more efficient and stable catalysts,careful design of the electrolytic cells for improving the mass transfer and development of environment-friendly processes for recovering the formed formate and biomass oxidation products.

关键词： Electrochemical reduction of CO_(2) Formic acid Oxidative conversion of biomass Lignocellulose Coupled system

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Using multiple site-directed modification of epoxide hydrolase to significantly improve its enantioselectivity in hydrolysis of rac-glycidyl phenyl ether

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Chinese Journal of Chemical Engineering 2020年第8期28卷 2181-2189页

作者： Yao Li Xiaoyang Ou Zewang Guo Minhua Zong Wenyong Lou Lab of applied biocatalysis School of Food Science and EngineeringSouth China University of TechnologyGuangzhou 510640China

The epoxide hydrolase gene(SpEH) from Sphingomonas sp. HXN-200 was synthesized and expressed in robust Escherichia coli cells that had a dual protection system. The enantioselectivity(E-value) of the recombinant SpEH was 7.7 and the yield of the remaining(R)-PGE was 24.3% for the hydrolysis of racemic phenyl glycidyl ether(rac-PGE). To improve the catalytic properties of SpEH, the site-directed mutagenesis was carried out based on homology modeling, sequence alignment and molecular docking. Six residues(V195, V196, F218,N226, Q312, and M332) near the active site were mutated to hydrophobic amino acids and the positive mutations were selected for combinatorial mutation. The optimal mutant SpEH^(V196A/N226A/M332A) had an enhanced E-value of 21.2 and a specific activity of 4.57 U·mg^-1-wet cells, which were 2.8-, and 2.3-fold higher than those of wild-type SpEH. The optimal temperature and p H for purified Sp EHV196 A/N226 A/M332 Ato catalyze the hydrolysis of rac-PGE were 25 ℃ and 7.0 with 200 U·mg^-1. The enantioselectivity and yield of the remaining(R)-PGE of E. coliSpEH^(V196A/N226A/M332A)increased from 7.7 to 21.2 and 24.3% to 40.9%, respectively. The molecular docking and kinetic parameter analyses showed that SpEH^(V196A/N226A/M332A) has a greater affinity toward(S)-PGE than(R)-PGE, and that it was more difficult for the O-atom of ASP170 to achieve the nucleophilic attack on the Cα of(R)-PGE, resulting in its improved enantioselectivity.

关键词： Epoxide hydrolase Phenyl glycidyl ether Enantioselectivity Directed modification

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Effect of Ionic Liquids on Catalytic Characteristics of Horse Liver Alcohol Dehydrogenase

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Chinese Journal of Chemistry 2006年第11期24卷 1643-1647页

作者：石贤爱宗敏华娄文勇 laboratory of applied biocatalysis South China University of Technology Guangzhou Guangdong 510640 China College of Biological Sciences & Technology Fuzhou University Fuzhou Fujian 350002 China

The catalytic characteristics of horse liver alcohol dehydrogenase （HLADH） in the systems involving ionic liquids （ILs）（BMIm·Cl, BMIm·Br, BMIm·PF6, BMIm·BF4 BMIm·OTf and EMIm·Cl） were examined. HLADH displayed higher oxidation activity towards ethanol in the systems containing BMIm·Cl, BMIm·Br, EMIm·Cl or BMIm·PF6 with proper content than that in the IL-free buffer. An excessive amount of these ILs in the reaction systems resulted in an obvious decline in enzymatic activity. BMIm·BF4 and BMIm·OTf of any content investigated could considerably inhibit the enzyme. The anions of ILs showed significant effect on the activity, kinetic parameters and activation energy of HLADH-mediated ethanol oxidation. Additionally, BMIm·Cl, BMIm·Br, EMIm·Cl and BMIm·PF6 boosted markedly the thermostability of HLADH, while the enzyme was less thermostable in BMIm·BF4 or BMIm·OTf-containing systems. The associated conformational changes in HLADH caused by ILs were examined by UV technique.

关键词： catalytic characteristics horse fiver alcohol dehydrogenase ionic liquid

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Metabolic engineering of an industrial bacterium Zymomonas mobilis for anaerobic l-serine production

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Synthetic and Systems Biotechnology 2024年第2期9卷 349-358页

作者： Zhen Wang Xia Wang Xiongying Yan Haixia Yi Shuche He Haoyu Zhang Xinli Zhou Qiaoning He Shihui Yang State Key laboratory of biocatalysis and Enzyme Engineering and School of Life Sciences Hubei UniversityWuhan430062China

Due to the complicated metabolic and regulatory networks of l-serine biosynthesis and degradation,microbial cell factories for l-serine production using non-model microorganisms have not been *** this study,a combination of synthetic biology and process optimization were applied in an ethanologenic bacterium Zymomonas mobilis for l-serine *** blocking the degradation pathway while introducing an exporter EceamA from ***,l-serine titer in recombinant *** was increased from 15.30 mg/L to 62.67 mg/*** was further increased to 260.33 mg/L after enhancing the l-serine biosynthesis ***,536.70 mg/L l-serine was achieved by removing feedback inhibition with a SerA mutant,and an elevated titer of 687.67 mg/L was further obtained through increasing serB copies while enhancing the ***,855.66 mg/L l-serine can be accumulated with the supplementation of the glutamate *** work thus not only constructed an l-serine producer to help understand the bottlenecks limiting l-serine production in *** for further improvement,but also provides guidance on engineering non-model microbes to produce biochemicals with complicated pathways such as amino acids or terpenoids.

关键词： Zymomonas mobilis Metabolic engineering l-serine Feedback inhibition Anaerobic fermentation

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Biosensor-assisted CRISPRi high-throughput screening to identify genetic targets in Zymomonas mobilis for high d-lactate production

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Synthetic and Systems Biotechnology 2024年第2期9卷 242-249页

作者： Qiqun Peng Weiwei Bao Binan Geng Shihui Yang State Key laboratory of biocatalysis and Enzyme Engineering and School of Life SciencesHubei UniversityWuhan430062China

Lactate is an important monomer for the synthesis of poly-lactate(PLA),which is a substitute for the petrochemical *** achieve the goal of high lactate titer,rate,and yield for commercial production,efficient lactate production pathway is needed as well as genetic targets that affect high lactate production and *** this study,an LldR-based d-lactate biosensor with a broad dynamic range was first applied into Zymomonas mobilis to select mutant strains with strong GFP fluorescence,which could be the mutant strains with increased d-lactate ***,LldR-based d-lactate biosensor was combined with a genome-wide CRISPR interference(CRISPRi)library targeting the entire genome to generate thousands of mutants with gRNA targeting different genetic targets across the whole ***,two mutant libraries were selected containing 105 and 104 mutants with different interference sites from two rounds of fluorescence-activated cell sorting(FACS),*** genetic targets of ZMO1323 and ZMO1530 were characterized and confirmed to be associated with the increased d-lactate production,further knockout of ZMO1323 and ZMO1530 resulted in a 15%and 21%increase of d-lactate production,*** work thus not only established a high-throughput approach that combines genome-scale CRISPRi and biosensor-assisted screening to identify genetic targets associated with d-lactate production in ***,but also provided a feasible high-throughput screening approach for rapid identification of genetic targets associated with strain performance for other industrial microorganisms.

关键词： D-lactate Biosensor LldR Genome-wide CRISPRi FACS Zymomonas mobilis

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Identifying RNA Modifications by Direct RNA Sequencing Reveals Complexity of Epitranscriptomic Dynamics in Rice

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Genomics, Proteomics & Bioinformatics 2023年第4期21卷 788-804页

作者： Feng Yu Huanhuan Qi Li Gao Sen Luo Rebecca Njeri Damaris Yinggen Ke Wenhua Wu Pingfang Yang State Key laboratory of biocatalysis and Enzyme Engineering School of Life SciencesHubei UniversityWuhan 430062China

Transcriptome analysis based on high-throughput sequencing of a cDNA library has been widely applied to functional genomic ***,the cDNA dependence of most RNA sequencing techniques constrains their ability to detect base modifications on RNA,which is an important element for the post-transcriptional regulation of gene *** comprehensively profile the N^(6)-methyladenosine(m^(6)A)and N^(5)-methylcytosine(m5 C)modifications on RNA,direct RNA sequencing(DRS)using the latest Oxford Nanopore Technology was applied to analyze the transcriptome of six tissues in *** 94 million reads were generated,with an average length ranging from 619 nt to 1013 nt,and a total of 45,707 transcripts across 34,763 genes were *** profiles of transcripts at the isoform level were quantified among ***-wide mapping of m^(6)A and m5 C demonstrated that both modifications exhibited tissue-specific *** transcripts with m^(6)A modifications tended to be modified by m5 C,and the transcripts with modifications presented higher expression levels along with shorter poly(A)tails than transcripts without modifications,suggesting the complexity of gene expression *** Ontology analysis demonstrated that m^(6)A-and m5 C-modified transcripts were involved in central metabolic pathways related to the life cycle,with modifications on the target genes selected in a tissue-specific ***,most modified sites were located within quantitative trait loci that control important agronomic traits,highlighting the value of cloning functional *** results provide new insights into the expression regulation complexity and data resource of the transcriptome and epitranscriptome,improving our understanding of the rice genome.

关键词： Direct RNA sequencing Polyadenylated transcriptome N6-methyladenosine N5-methylcytosine Rice

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Transforming liquid flow fuel cells to controllable reactors for highlyefficient oxidation of 5-hydroxymethylfurfural to 2, 5-furandicarboxylic acid at low temperature

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Journal of Energy Chemistry 2024年第3期90卷 621-631,I0014页

作者： Ye Qiang Xi Liu Denghao Ouyang Zhao Jiang Xuebing Zhao Key laboratory of Industrial biocatalysis Ministry of EducationTsinghua UniversityBeijing 100084China Institute of applied Chemistry Department of Chemical EngineeringTsinghua UniversityBeijing 100084China School of Chemical Engineering and Technology Xi'an Jiaotong UniversityXi'an 710049ShaanxiChina

Highly-efficient oxidation of 5-hydroxymethylfurtural(HMF) to 2,5-furandicarboxylic acid(FDCA) at low temperature with air as the oxidant is still ***,inspired by the respirato ry electron transport chain(ETC) of living cells mediated by electron carriers,we constructed artificial ETCs and transformed liquid flow fuel cells(LFFCs) to flexible reactors for efficient oxidation of HMF to produce FDCA under mild *** LFFC reactor employed an electrodeposition modified nickel foam as an anode to promote HMF oxidation and(VO_(2))_(2)SO_(4) as a cathode electron carrier to facilitate the electron transfer to *** reaction rate could be easily controlled by selecting the anode catalyst,adjusting the external loading and changing the cathodic electron carrier or oxidants.A maximal power density of 44.9 mW cm^(-2) at room temperature was achieved,while for FDCA production,short-circuit condition was preferred to achieve quick transfer of *** a single batch operation with 0.1 M initial HMF,FDCA yield reached 97.1%.By fed-batch operation,FDCA concentration reached 144.5 g L^(-1) with a total yield of 96%.Ni^(2+)/Ni^(3+) redox couple was the active species mediating the electron transfer,while both experimental and DFT calculation results indicated that HMFCA pathway was the preferred reaction mechanism.

关键词： 5-Hydroxymethylfurfural 2 5-Furandicarboxylic acid Electrodeposition Electron transport chain Liquid flow fuel cell

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Advancing organelle genome transformation and editing for crop improvement

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Plant Communications 2021年第2期2卷 16-28页

作者： Shengchun Li Ling Chang Jiang Zhang State Key laboratory of biocatalysis and Enzyme Engineering School of Life SciencesHubei UniversityWuhan 430062China

Plant cells contain three organelles that harbor DNA:the nucleus,plastids,and *** transformation has emerged as an attractive platformfor the generation of transgenic plants,also referred to as transplastomic *** genomes have been genetically engineered to improve crop yield,nutritional quality,and resistance to abiotic and biotic stresses,as well as for recombinant protein *** many promising proof-of-concept applications,transplastomic plants have not been commercialized to ***-specific nuclease technologies are widely used to precisely modify nuclear genomes,but these tools have not been applied to edit organelle genomes because the efficient homologous recombination system in plastids facilitates plastid genome *** plastid transformation,successful genetic transformation of higher plant mitochondrial genome transformation was tested in several research group,but not successful to ***,stepwise progress has been made in modifying mitochondrial genes and their transcripts,thus enabling the study of their ***,we provide an overview of advances in organelle transformation and genome editing for crop improvement,and we discuss the bottlenecks and future development of these technologies.

关键词： organelle transformation genome editing homologous recombination crop improvement

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Promoting and controlling electron transfer of furfural oxidation efficiently harvest electricity,furoic acid,hydrogen gas and hydrogen peroxide

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Journal of Energy Chemistry 2023年第4期79卷 135-147页

作者： Denghao Ouyang Daihong Gao Jinpeng Hong Zhao Jiang Xuebing Zhao Key laboratory of Industrial biocatalysis Ministry of EducationTsinghua UniversityBeijing 100084China Institute of applied Chemistry Department of Chemical EngineeringTsinghua UniversityBeijing 100084China School of Chemical Engineering and Technology Xi’an Jiaotong UniversityXi’an 710049ShaanxiChina

Conventional chemical oxidation of aldehydes such as furfural to corresponding acids by molecular oxygen usually needs high pressure to increase the solubility of oxygen in aqueous phase,while electrochemical oxidation needs input of external electric ***,we developed a liquid flow fuel cell(LFFC)system to achieve oxidation of furfural in anode for furoic acid production with co-production of hydrogen *** controlling the electron transfer in cathode for reduction of oxygen,efficient generation of electricity or production of H_(2)O_(2)were *** oxides especially Ag_(2)O have been screened as the efficient catalyst to promote the oxidation of aldehydes,while liquid redox couples were used for promoting the kinetics of oxygen reduction.A novel alkaline-acidic asymmetric design was also used for anolyte and catholyte,respectively,to promote the efficiency of electron *** an LFFC system achieves efficient conversion of chemical energy of aldehyde oxidation to electric energy and makes full use the transferred electrons for high-value added products without input of external ***(VO_(2))_(2)SO_(4)as the electron carrier in catholyte for four-electron reduction of oxygen,the peak output power density(Pmax)at room temperature reached 261 mW/cm^(2)with furoic acid and H_(2)yields of 90%and 0.10 mol/mol furfural,*** anthraquinone-2-sulfonate(AQS)as the cathodic electron carrier,Pmaxof 60 mW/cm^(2)and furoic acid,H_(2)and H_(2)O_(2)yields of 0.88,0.15 and 0.41 mol/mol furfural were achieved,respectively.A new reaction mechanism on furfural oxidation on Ag_(2)O anode was proposed,referring to one-electron and two-electron reaction pathways depending on the fate of adsorbed hydrogen atom transferred from furfural aldehyde group.

关键词： Oxidation of furfural Liquid flow fuel cell Electricity generation Hydrogen production Electron transfer

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