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Green Synthesis and Catalysis 2024年第2期5卷 80-87页

作者： Yongxing Li Pengqian Luan Lele Dong Jianqiao Liu Luying Jiang Jing Bai Fufeng Liu Yanjun Jiang School of Chemical Engineering and Technology Hebei University of TechnologyTianjin 300130China College of Food Science and Biology Hebei University of Science&Technology26 Yuxiang StreetYuhua DistrictShijiazhuang 050018China Key laboratory of industrial Fermentation microbiology of Ministry of Education Tianjin Key Laboratory of Industrial MicrobiologyCollege of BiotechnologyTianjin University of Science&TechnologyTianjin 300457China

Asymmetric reduction of the conjugated C=C bonds by the old yellow enzymes(OYEs)presents a promising field in the synthesis of chiral ***,few natural OYEs have been applied in large-scale applications due to the requirement of costly NADPH and low operational ***,a stable and efficient fusion of YqjM from Bacillus subtilis and glucose dehydrogenase(GDH)from Bacillus megaterium was constructed to stereoselectively reduce the conjugated C=C bonds in a self-sufficient continuous *** effects of the enzyme order and different linkers on the fusions were investigated by structural analysis and all-atom molecular dynamics *** best fusion YqjM_G_GDH gave 98% conversion of 100 mmol/L 2-methylcyclopentenone with an excellent ee value(>99%)in 3 h,while the mixture of individual enzymes only obtained 68% conversion after more than 8 *** improved substrate conversion of YqjM_G_GDH fusion was probably attributed to the increased flexibility of each fused enzyme and the shortening of the diffusion distance of NADPH regenerated.A one-pot process was designed to purify and immobilize the fusion on the Ni2t-nitrilotriacetic acid functionalized magnetic mesoporous silica *** resulting immobilized biocatalyst not only catalyzed the asymmetric reduction of various α,β-unsaturated ketones(20 mmol/L)continuously with only 50μmol/L NADPt to initiate the whole process,but also retained more than 82%of the initial activity after seven cycles,serving as a good candidate for the industrial applications.

关键词： Enzyme fusion Old yellowenzymes Asymmetricreduction C=C bond Cofactor regeneration Immobilization

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Lindqvist-type Polyoxometalates Act as Anti-breast Cancer Drugs via Mitophagy-induced Apoptosis

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Current Medical Science 2024年第4期44卷 809-819页

作者： Chen-guang YAO Zi-jia ZHAO Ting TAN Jiang-ning YAN Zhong-wei CHEN Jun-tao XIONG Han-luo LI Yan-hong WEI Kang-hong HU Sino-German Biomedical Center Hubei Provincial Key Laboratory of Industrial MicrobiologyCooperative Innovation Center of Industrial Fermentation(Ministry of Education&Hubei Province)Key Laboratory of Fermentation Engineering(Ministry of Education)National“111”Center for Cellular Regulation and Molecular PharmaceuticsHubei University of TechnologyWuhan430068China Center for Evaluation of Hubei Medical Products Administration Wuhan430068China

Objective Lindqvist-type polyoxometalates(POMs)exhibit potential antitumor *** study aimed to examine the effects of Lindqvist-type POMs against breast cancer and the underlying *** Using different cancer cell lines,the present study evaluated the antitumor activities of POM analogues that were modified at the body skeleton based on molybdenum-vanadium-centered negative oxygen ion polycondensations with different side *** colony formation assay,autophagy detection,mitochondrial observation,qRT-PCR,Western blotting,and animal model were used to evaluate the antitumor activities of POMs against breast cancer cells and the related *** MO-4,a Lindqvist-type POM linking a proline at its side strain,was selected for subsequent experiments due to its low half maximal inhibitory concentration in the inhibition of proliferation of breast cancer *** was found that MO-4 induced the apoptosis of multiple types of breast cancer ***,MO-4 activated intracellular mitophagy by elevating mitochondrial reactive oxygen species(ROS)levels and resulting in *** vivo,breast tumor growth and distant metastasis were significantly reduced following MO-4 *** Collectively,the results of the present study demonstrated that the novel Lindqvist-type POM MO-4 may exhibit potential in the treatment of breast cancer.

关键词： antitumor breast cancer polyoxometalate mitochondria mitophagy

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Integrated pathway engineering and transcriptome analysis for improved astaxanthin biosynthesis in Yarrowia lipolytica

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Synthetic and Systems Biotechnology 2022年第4期7卷 1133-1141页

作者： Dan-Ni Wang Jie Feng Chen-Xi Yu Xin-Kai Zhang Jun Chen Liu-Jing Wei Zhijie Liu Liming Ouyang Lixin Zhang Qiang Hua Feng Liu State Key laboratory of Bioreactor Engineering East China University of Science and Technology130 Meilong RoadShanghai200237PR China Key laboratory of Fermentation Engineering(Ministry of Education) Cooperative Innovation Center of Industrial Fermentation(Ministry of Education&Hubei Province)Hubei Key Laboratory of Industrial MicrobiologyHubei University of TechnologyWuhan430068China Shanghai Collaborative Innovation Center for Biomanufacturing Technology 130 Meilong RoadShanghai200237China

Astaxanthin is a high value carotenoid with a broad range of commercial applications due to its superior antioxidant *** this study,β-carotene-producing Yarrowia lipolytica XK17 constructed in the lab was employed for astaxanthin *** catalytic effects ofβ-carotene ketolase CrtW andβ-carotene hydroxylase CrtZ from various species were *** PspCrtW from Paracoccus *** HpCrtZ^(#) from Haematococcus pluvialis were confirmed to be the best combination in convertingβ-*** key bottlenecks in biomass and astaxanthin biosynthesis were effectively eliminated by optimizing the expression of the above enzymes and restoring uracil/leucine *** addition,the effects of astaxanthin biosynthesis on cell metabolism were investigated by integrated analysis of pathway modification and transcriptome *** further optimization,strain DN30 was able to synthesize up to 730.3 mg/L astaxanthin in laboratory 5-L *** study provides a good metabolic strategy and a sustainable development platform for high-value carotenoid production.

关键词： Astaxanthin Pathway engineering Transcriptome analysis Yarrowia lipolytica

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Platycladus orientalis shells promote melanogenesis via p38,AKT and Wnt/β-catenin signaling pathways

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Traditional Medicine Research 2022年第6期7卷 56-64页

作者： Fu-Rui Liu Hao Xiang Qian-Bei Guo Shu-Li Man Long Ma State Key laboratory of Food Nutrition and Safety Key Laboratory of Industrial MicrobiologyMinistry of EducationTianjin Key Laboratory of Industry MicrobiologyNational and Local United Engineering Lab of Metabolic Control Fermentation TechnologyChina International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal ChemistryCollege of BiotechnologyTianjin University of Science&TechnologyTianjin 300457China

Background:Platycladus orientalis,which has been employed in traditional Chinese medicine for cool blood,antibacterial,promotion of hair growth and therapy of poliosis for ***,there have been few reports focusing on Platycladus orientalis shells treating pigmentary ***:In present study,gas chromatography–mass spectrometry was applied to analyzing the volatile composition of Platycladus orientalis shells and cellular *** in vivo were carried out to evaluate the effect of Platycladus orientalis shells treating pigmentation disorders in C57BL/6J ***:Our results indicated that cedrol occupied the largest percentage in Platycladus orientalis shells(17.1%).Meanwhile,Platycladus orientalis shells up-regulated the content of palmitic acid and increased melanin content and tyrosinase *** mechanism possibly involved in the inhibiting phosphorylation of AKT and β-catenin,increasing phosphorylation of p38 to promote microphthalmia-associated transcription factor *** animal experiment also proved that Platycladus orientalis shells promoted melanogensis and hair blacken in C57BL/6J ***:All in all,Platycladus orientalis shells potentially promoted melanogenesis in vitro and in *** was regarded as the main active substance in Platycladus orientalis ***,it could be used for treatment of pigmentary disorders under safe concentration in the prospective application.

关键词： GC-MS Platycladus orientalis shells melanogenesis tyrosinase pigmentary disorders

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Rhizoma paridis saponins protected against liver injury in diethylnitrosamine-induced mice

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Traditional Medicine Research 2023年第2期8卷 7-12页

作者： Chen Luo Jing-Wen Yao Shu-Li Man Wen-Yuan Gao Key laboratory of industrial microbiology Ministry of Education/Tianjin Key Laboratory of Industry Microbiology/National and Local United Engineering Lab of Metabolic Control Fermentation Technology/College of BiotechnologyTianjin University of Science&TechnologyTianjin 300457China Tianjin Key laboratory for Modern Drug Delivery and High Efficiency School of Pharmaceutical Science and TechnologyTianjin UniversityTianjin 300072China

Background:Diethylnitrosamine,one of food additives,possessed a strong carcinogenic effect in *** paridis saponins,as the main active components of Paris polyphylla,have a good anti-cancer effect in our previous *** verify their inhibitory effect on diethylnitrosamine-induced liver cancer,we carried out this ***:We established diethylnitrosamine-induced mouse hepatocarcinoma models to evaluate antitumor of Rhizoma paridis ***,gas chromatography-mass spectrometry was applied to analyze the metabolites in the urine and serum ***:Rhizoma paridis saponins alleviated diethylnitrosamine-induced *** the one hand,Rhizoma paridis saponins down-regulated the levels of liver function markers,such as alanine aminotransferase,aspartate transaminase and alpha *** the other hand,Rhizoma paridis saponins reduced metabolic disorders by increasing fructose and mannose metabolism,and decreasing pentose and glucuronate interconversion,inositol phosphate metabolism,and the process of saturated fatty acids transforming to unsaturated fatty acids,which based on the regulating mRNA expression of glucose transporter type 4,lactate dehydrogenase A,fatty acid synthetas,acetyl-CoA carboxylase and apolipoprotein ***:Rhizoma paridis saponins has the potential application to inhibit chemical-induced hepatocarcinogenesis in the future.

关键词： Rhizoma paridis saponins liver injury diethylnitrosamine metabolites

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Distribution and expression in vitro and in vivo of DNA vaccine against lymphocystis disease virus in Japanese flounder (Paralichthys olivaceus)

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Chinese Journal of Oceanology and Limnology 2010年第1期28卷 67-74页

作者：郑风荣孙修勤刘洪展吴兴安钟楠王波周国栋 First Institute of Oceanography SOA Marine College Shandong University at Weihai Department of microbiology Fourth Military Medical University Hainan Green Food industrial company Ltd

Lymphocystis disease,caused by the lymphocystis disease virus (LCDV),is a significant worldwide problem in fish industry causing substantial economic *** this study,we aimed to develop the DNA vaccine against LCDV,using DNA vaccination *** evaluated plasmid pEGFP-N2-LCDV1.3 kb as a DNA vaccine *** plasmid DNA was transiently expressed after liposome transfection into the eukaryotic COS 7 cell *** distribution and expression of the DNA vaccine (pEGFP-N2-LCDV1.3kb) were also analyzed in tissues of the vaccinated Japanese flounder by PCR,RT-PCR and fluorescent *** from PCR analysis indicated that the vaccine-containing plasmids were distributed in injected muscle,the muscle opposite the injection site,the hind intestine,gill,spleen,head,kidney and liver,6 and 25 days after *** vaccine plasmids disappeared 100 d *** microscopy revealed green fluorescence in the injected muscle,the muscle opposite the injection site,the hind intestine,gill,spleen,head,kidney and liver of fish 48 h post-vaccination,green fluorescence did not appear in the control treated *** fluorescence became weak at 60 days ***-PCR analysis indicated that the mcp gene was expressed in all tested tissues of vaccinated fish 6-50 days *** results demonstrate that the antigen encoded by the DNA vaccine is distributed and expressed in all of the tissues analyzed in the vaccinated *** antigen would therefore potentially initiate a specific immune *** plasmid DNA was injected into Japanese flounder (Paralichthys olivaceus) intramuscularly and antibodies against LCDV were *** results indicate that the plasmid encoded DNA vaccine could induce an immune response to LCDV and would therefore offer immune protection against *** studies are required for the development and application of this promising DNA vaccine.

关键词： DNA vaccine lymphocystis disease virus Paralichthys olivaceus

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Single carbon metabolism–A new paradigm for microbial bioprocesses?

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Synthetic and Systems Biotechnology 2024年第2期9卷 322-329页

作者： Michael Baumschabl Özge Ata Diethard Mattanovich Austrian Centre of industrial Biotechnology(ACIB) Vienna1190Austria University of Natural Resources and Life Sciences Department of BiotechnologyInstitute of Microbiology and Microbial BiotechnologyViennaAustria

Increasing atmospheric carbon dioxide levels,a reduction of arable land area and the dependence of first and second generation biotechnology feedstocks on agricultural products,call for alternative,sustainable feedstock sources for industrial *** direct use of CO_(2)or conversion of CO_(2)into other single carbon(C1)sources have great potential as they might help to reduce carbon emissions and do not compete with agricultural land *** we discuss the microbial use of C1 carbon sources,their potential applications in biotechnology,and challenges towards sustainable C1-based industrial biotechnology *** focus on methanol,formic acid,methane,syngas,and CO_(2)as feedstocks for bioprocesses,their assimilation pathways,current and emerging applications,and limitations of their *** mini-review is intended as a first introduction for researchers who are new to the field of C1 biotechnology.

关键词： Sustainability BioeconomyCarbon dioxide Methanol Formate Methane

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Purification and biochemical characterization of manganesecontaining superoxide dismutase from deep-sea thermophile Geobacillus sp. EPT3

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Acta Oceanologica Sinica 2014年第12期33卷 163-169页

作者： ZHU Yanbing LI Hebin NI Hui LIU Jingwen XIAO Anfeng CAI Huinong College of Food and Biological Engineering Jimei University Fujian Provincial Key laboratory of Food microbiology and Enzyme Engineering Research Center of Food microbiology and Enzyme Engineering Technology of Fujian Province Xiamen Medical College

Thermostable SOD is a promising enzyme in biotechnological applications. In the present study, ***3 was isolated from a deep-sea hydrothermal field in the East Pacific. A thermo-stable superoxide dismutase （SOD） from this strain was purified to homogeneity by steps of fractional am-monium sulfate precipitation, DEAE-Sepharose chromatography, and Phenyl-Sepharose chromatography. SOD was purified 13.4 fold to homogeneity with a specific activity of 3 354 U/mg and 11.1% recovery. SOD fromGeobacillussp. EPT3 was of the Mn-SOD type, judged by the insensitivity of the enzyme to both KCN and H2O2. SOD was determined to be a homodimer with monomeric molecular mass of 26.0 kDa. It had high thermostability at 50＆#176;C and 60＆#176;C. At tested conditions,SOD was relatively stable in the presence of some inhibitors and denaturants, such asβ-mercaptoethanol （β-ME）, dithiothreitol （DTT）, phenylmethylsulfonyl fluoride （PMSF）, urea, and guanidine ***. EPT3 SOD showed striking stability across a wide pH range from 5.0 to 11.0. It could withstand denaturants of extremely acidic and alkaline conditions, which makes it useful in the industrial applications.

关键词： Geobacillus sp. superoxide dismutase thermostability purification

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Purification and Characterization of an Antibacterial Protein from the Cultured Mycelia of Cordyceps sinensis

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Wuhan University Journal of Natural Sciences 2006年第3期11卷 709-714页

作者： HU Zheng YE Maoqing XIA Liqiu TU Wenjuan LI Liang ZOU Guolin College of Life Sciences Wuhan University Wuhan 430072Hubei China Key laboratory of industrial microbiology of Hubei Province Hubei University of Technology Wuhan 430068 Hubei China College of Life Sciences Hunan Normal University Changsha 410081 Hunan China

An antibacterial protein was isolated from the cultured mycelia of Cordyceps sinensis, and was designated as Cordyceps sinensis Antibacterial Protein （CSAP）. CSAP was single-chained, with an apparent molecular mass of 35 × 10^3 revealed by SDS-PAGE and a novel hydrophobic N-terminal sequence N-ALATQHGAP. The antimicrobial assays showed CSAP could inbibit the growth of Gram-positive and Gram-negative bacteria but no significant inhibition against fungi or yeasts. Further more, the antibacterial activity of CSAP was not bactericidal but bacteriostatic. It was the first time that an antibacterial protein was described in the Cordyceps species, which might involve in the chemical defense mechanism of the hosts.

关键词： Cordyceps species antimicrobial protein isolation identification membrane-activity proteins

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Rational design for fungal laccase production in the model host Aspergillus nidulans

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Science China(Life Sciences) 2019年第1期62卷 84-94页

作者： Wei Li Jingwen Yu Zixin Li Wen-Bing Yin State Key laboratory of Mycology Institute of Microbiology Chinese Academy of Sciences Savaid Medical School University of Chinese Academy of Sciences

Laccases, multicopper oxidoreductases, are mainly produced in white-rot fungi and are considered as ideal green catalysts in industrial and biotechnological applications. However, the development of laccases is limited due to the slow growth of natural laccase producing strains and the low expression levels of laccases. In this study, we designed three regulation strategies for laccase gene expression in the model fungus Aspergillus nidulans. By introducing various promoters in front of the laccase gene pslcc from the white-rot fungus Pycnoporus sanguineus, we found that the laccase gene with the original promoter had effective expression in A. nidulans. Using the previously identified transcription factor RsmA regulatory mechanism, the aflR promoter was inserted into the pslcc expression vectors, and the laccase production was 15-fold higher in the strain overexpressing of RsmA compared to the control strain. To improve the laccase yield, the dipeptidyl-peptidase DppV, aspartic protease PepA and mannosyltransferase Mnn9 were successfully deleted in the A. nidulans host. The laccase activities were increased approximately 8-fold and 13-fold in the double deletions strains of Δmnn9ΔpepA and ΔdppVΔpepA over the control strains, *** together, these results not only demonstrate an efficient system for heterologous protein production in the model fungus *** but also provide a general approach to applying regulatory methods to control gene expression.

关键词： laccase heterologous expression promoter Aspergillus nidulans

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