作者： Chen, Si;Feng, Hao;Li, Xin;Chao, Hong-jun;Wu, Jing;...

期刊： Current Microbiology,2020年77(12):3945-3952 ISSN：0343-8651

通讯作者： Yan, Da-zhong

作者机构： [Wu, Jing; Chao, Hong-jun; Yan, Da-zhong; Chen, Si; Zhu, Wen-jun; Liu, Jun; Li, Xin] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan 430023, Peoples R China.;[Feng, Hao] Jiangsu Yanghe Brewery Joint Stock Co Ltd, Suqian 223800, Jiangsu, Peoples R China.

通讯机构： [Yan, Da-zhong] W;Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan 430023, Peoples R China.

摘要： Many organisms secrete xylanase, an import group of proteins hydrolyzing xylan, and thus are able to use xylan as their carbon source. In this study, we sequenced the whole genome of a bacterial strain, YD01, which was isolated from the sludge near the sewage discharge outlet of a papermill and showed high alkalic xylanase activity. Its genome consists of a chromosome and two plasmids. Six rRNA genes, 46 tRNA genes, 3136 CDSs as well as 955 repetitive sequences were predicted. 3046 CDSs were functionally annotated. Phylogenetic analysis on 16S rRNA shows that YD01 is a new species in Microbacterium genus and is taxonomically close to M. jejuense THG-C31T and M. kyungheense THG-C26T. A comparative study on phylogenetic trees of 16S rRNA and xylanase genes suggests that xylanase genes in YD01 may originate from horizontal gene transfer instead of ancestral gene duplication. © 2020, Springer Science+Business Media, LLC, part of Springer Nature.

语种： 英文

作者： Chen, Yuanyuan;Li, Yan;Chao, Hongjun;Wu, Jing;Zhu, Wenjun;...

期刊： Process Biochemistry,2020年91:65-72 ISSN：1359-5113

通讯作者： Yan, Dazhong

作者机构： [Wu, Jing; Yan, Dazhong; Li, Yan; Chen, Yuanyuan; Zhu, Wenjun; Chao, Hongjun] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, 68 Xuefu South Rd, Wuhan 430023, Peoples R China.;[Fang, Ti] Chinese Acad Sci, Wuhan Inst Virol, State Key Lab Virol, 44 Xiaohongshan, Wuhan 430071, Peoples R China.;[Gao, Xuewang] Chinese Acad Sci, Tech Inst Phys & Chem, Beijing 100190, Peoples R China.

通讯机构： [Yan, Dazhong] W;Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, 68 Xuefu South Rd, Wuhan 430023, Peoples R China.

关键词： Cellulosimicrobium cellulans ATCC21606;Pseudomonas putida PaW340;Recombinant expression;Thermostability;Xanthine oxidase

摘要： Xanthine oxidase (XOD) catalyses the oxidation of hypoxanthine into xanthine and xanthine into uric acid. The enzyme plays a key role in the purine metabolic pathway. Despite the presence of different XODs in prokaryotes, the functional and structural knowledge of prokaryotic XODs remain limited (compared with their well-known eukaryotic counterparts), thereby hindering their biochemical analysis and industrial application. Using genetic and biochemical analyses, we identified and characterised recombinant XOD (CcXODAB) from Cellulosimicrobium cellulans ATCC21606. Bioinformatics analysis suggests that CcXODAB shares low amino acid sequence identities with other XODs. The purified enzyme exhibits the maximum activity at 55 °C and pH 8.0. In addition, CcXODAB exhibits moderate thermostability and retains 80.65 % of the original activity after 30 min of incubation at 60 °C. Ca2 + has a slight inhibitory effect, whereas Co2 + and Mn2 + have a strong inhibitory effect on XODAB activity. In particular, low Ba2+ and Mg2 + concentrations have no effect, whereas high Mg2 + (≥10 mM) and Ba2+ (≥2 mM) concentrations show an inhibitory effect on enzyme activity. The Km and Vmax values for xanthine are 131.29 ± 11.09 μmol•L−1 and 15.23 ± 0.65 μmol•L-1 min−1, respectively. Results indicate that CcXODAB is a novel enzyme with potential industrial application. © 2019

语种： 英文

作者： Liu, Jun;Han, Qin;Cheng, Qikun;Chen, Yuanyuan;Wang, Ruxin;...

期刊： Current Microbiology,2020年77(5):846-854 ISSN：0343-8651

通讯作者： Yan, Dazhong

作者机构： [Liu, Yulan; Yan, Dazhong; Liu, Jun] Wuhan Polytech Univ, Hubei Key Lab Anim Nutr & Feed Sci, Wuhan, Peoples R China.;[Yan, Dazhong; Wang, Ruxin; Chen, Yuanyuan; Cheng, Qikun; Han, Qin; Liu, Jun; Li, Xin] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan, Peoples R China.

通讯机构： [Yan, Dazhong] W;Wuhan Polytech Univ, Hubei Key Lab Anim Nutr & Feed Sci, Wuhan, Peoples R China.;Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan, Peoples R China.

摘要： In this work, the high-level expression of the human lysozyme (HLY) was achieved by both optimization of the gene copy number and co-expression of the transcription factor Hac1p for the unfolded protein response (UPR) in the host strain Pichia pastoris KM71H. A series of recombinant constructs with various numbers of HLY expression cassettes was generated for the production of recombinant strains integrated with different copies of the HLY gene. The copy number of the HLY gene was determined by real-time quantitative polymerase chain reaction, and the recombinant strains of P. pastoris carrying one, two, three, four, or six copies of the HLY gene were obtained. Maximum extracellular protein and lysozyme enzyme activity reached 436.99 ± 26.08μg/mL and 61,900 ± 2036.47 U/mL, respectively, in the recombinant strain HLYH4-3 with the four copies of the HLY gene after shaking flask fermentation. Moreover, the co-expression of the transcription factor Hac1p in the recombinant strains further enhanced the HLY yields. Extracellular protein and lysozyme enzyme activity, respectively, reached 517.82 ± 4.19μg/mL and 78,600 ± 1134.95 U/mL by using the Hac1p co-expression strain HLYH4-3/Hac1p. These values are the highest recorded level of human lysozyme expressed by P. pastoris in shaking flask fermentation so far. © 2020, Springer Science+Business Media, LLC, part of Springer Nature.

语种： 英文

作者： Huang, Li;Luo, Dehua;Gondil, Vijay S.;Gong, Yujing;Jia, Minghui;...

期刊： Applied Microbiology and Biotechnology,2020年104(4):1609-1619 ISSN：0175-7598

通讯作者： Hu, Shencai;Yang, Hang;Wei, Hongping

作者机构： [Huang, Li; Yan, Dazhong; Hu, Shencai] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan 430023, Peoples R China.;[Luo, Dehua; Gong, Yujing; He, Jin; Jia, Minghui] Huazhong Agr Univ, Coll Life Sci & Technol, State Key Lab Agr Microbiol, Wuhan 430070, Peoples R China.;[Yang, H; Wei, Hongping; Yang, Hang; Gondil, Vijay S.] Chinese Acad Sci, Ctr Emerging Infect Dis, Wuhan Inst Virol, CAS Key Lab Special Pathogens & Biosafety, Wuhan 430071, Peoples R China.

通讯机构： [Hu, Shencai] W;[Yang, H; Wei, HP] C;Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan 430023, Peoples R China.;Chinese Acad Sci, Ctr Emerging Infect Dis, Wuhan Inst Virol, CAS Key Lab Special Pathogens & Biosafety, Wuhan 430071, Peoples R China.

关键词： Bacteriophage lysin;Chimeric lysin;GBS;Therapeutics;Susceptibility

摘要： The emergence of antibiotic-resistant beta-hemolytic Streptococcus agalactiae strains poses increasing threat to human beings globally. As an attempt to create a novel lysin with improved activity against S. agalactiae, a chimeric lysin, ClyV, was constructed by fusing the enzymatically active domain (EAD) from PlyGBS lysin (GBS180) and the cell wall binding domain (CBD) from PlyV12 lysin (V12CBD). Plate lysis assay combined with lytic kinetic analysis demonstrated that ClyV has improved activity than its parental enzymatic domain GBS180 against multiple streptococci. Biochemical characterization showed that ClyV is active from pH7 to 10, with the optimum pH of 9, and is stable under NaCl concentration of < 500mM. In a S. agalactiae infection model, a single intraperitoneally administration of 0.1mg/mouse of ClyV protected 100% mice, while it was observed that ~ 29% survive in group that received a single dose of 0.1mg/mouse of GBS180. Moreover, a high dose of 0.8mg/mouse ClyV did not show any adverse effects to the health or survival rate of the mice. Considering the robust bactericidal activity and good safety profile of ClyV, it represents a potential candidate for the treatment of S. agalactiae infections. © 2020, Springer-Verlag GmbH Germany, part of Springer Nature.

语种： 英文

作者： Chao, Hong-Jun;Chen, Yuan-yuan;Wu, Jing;Yan, Da-Zhong*;Zhou, Ning-Yi

期刊： Current Microbiology,2019年76(11):1235-1237 ISSN：0343-8651

通讯作者： Yan, Da-Zhong

作者机构： [Wu, Jing; Chao, Hong-Jun; Chen, Yuan-yuan; Yan, Da-Zhong] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan, Hubei, Peoples R China.;[Chao, Hong-Jun; Zhou, Ning-Yi] Chinese Acad Sci, Wuhan Inst Virol, Wuhan, Hubei, Peoples R China.;[Zhou, Ning-Yi] Shanghai Jiao Tong Univ, State Key Lab Microbial Metab, Shanghai, Peoples R China.;[Zhou, Ning-Yi] Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai, Peoples R China.

通讯机构： [Yan, Da-Zhong] W;Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan, Hubei, Peoples R China.

摘要： Chlorobenzenes are ubiquitously distributed, highly persistent, and toxic environmental contaminants. Pandoraea pnomenusa MCB032 was isolated as a new dominant chlorobenzene-utilizing strain from a functionally stable bioreactor during the treatment of chlorobenzenes when strain Burkholderia sp. JS150 disappeared. In study, we report the complete genome sequence of strain MCB032 which consists of a circular chromosome and three plasmids, which are ~ 6Mb in length with 5450 open reading frames—12 encoding rRNAs and 77 encoding tRNAs. We further identified 17 putative genes encoding the enzymes involved in the methyl-accepting chemotaxis proteins in sensing chemical gradients during chemotaxis. The annotated complete genome sequence of this strain will provide genetic insights into the degradation of chlorinated aromatic compounds. The information will empower the elucidation of chlorobenzene affinity hierarchy and species succession in the bioreactor. © 2019, Springer Science+Business Media, LLC, part of Springer Nature.

语种： 英文

作者： Yan, Da-Zhong*;Gan, Ya-Ting;Zhou, Hui;Liu, Jun;Li, Xin

期刊： Current Microbiology,2019年76(9):1092-1092 ISSN：0343-8651

通讯作者： Yan, Da-Zhong

作者机构： [Zhou, Hui; Gan, Ya-Ting; Liu, Jun; Yan, Da-Zhong; Li, Xin] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, 68 Xuefu South Rd, Wuhan 430023, Hubei, Peoples R China.

通讯机构： [Yan, Da-Zhong] W;Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, 68 Xuefu South Rd, Wuhan 430023, Hubei, Peoples R China.

摘要： The original version of this article unfortunately contained a mistake in the Fig. S1 of supplementary material. It is corrected with this erratum.

语种： 英文

作者： Tao, Weixin*;Chen, Li;Zhao, Chunhua;Wu, Jing;Yang, Dazhong;...

期刊： ACS Synthetic Biology,2019年8(9):1991-1997 ISSN：2161-5063

通讯作者： Tao, Weixin;Sun, Yuhui

作者机构： [Sun, Yuhui; Tao, Weixin; Deng, Zixin; Chen, Li; Tao, WX; Sun, YH] Minist Educ, Key Lab Combinatorial Biosynth & Drug Discovery, Wuhan 430071, Hubei, Peoples R China.;[Sun, Yuhui; Tao, Weixin; Deng, Zixin; Chen, Li; Tao, WX; Sun, YH] Wuhan Univ, Sch Pharmaceut Sci, Wuhan 430071, Hubei, Peoples R China.;[Zhao, Chunhua] Huazhong Univ Sci & Technol, Tongji Med Coll, Sch Pharm, Wuhan 430030, Hubei, Peoples R China.;[Wu, Jing; Yang, Dazhong] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan 430023, Hubei, Peoples R China.

通讯机构： [Tao, WX; Sun, YH] M;[Tao, WX; Sun, YH] W;Minist Educ, Key Lab Combinatorial Biosynth & Drug Discovery, Wuhan 430071, Hubei, Peoples R China.;Wuhan Univ, Sch Pharmaceut Sci, Wuhan 430071, Hubei, Peoples R China.

关键词： direct cloning;natural product;in vitro packaging;CRISPR/Cas9;gene editing

摘要： Direct cloning of natural product pathways for efficient refactoring and heterologous expression has become an important strategy for microbial natural product research and discovery, especially for those kept silent or poorly expressed in the original strains. Accordingly, the development of convenient and efficient cloning approaches is becoming increasingly necessary. Here we presented an in vitro packaging mediated cloning approach that combines CRISPR/Cas9 system with in vitro λ packaging system, for targeted cloning of natural product pathways. In such a scheme, pathways of Tü3010 (27.4 kb) and sisomicin (40.7 kb) were respectively cloned, and stuR was further depicted to positively regulate Tü3010 production. In vitro packaging mediated approach not only enables to activate cryptic pathways, but also facilitates refactoring or interrogating the pathways in conjunction with various gene editing systems. This approach features an expedited, convenient, and generic manner, and it is conceivable that it may be widely adopted for targeted cloning of the natural product pathways. © 2019 American Chemical Society.

语种： 英文

作者： Zhu, Wenjun;Xu, Xiaowen;Peng, Fang;Yan, Da-zhong;Zhang, Shaopeng;...

期刊： Plant Science,2019年283:1-10 ISSN：0168-9452

通讯作者： Wei, Wei;Chen, Weidong

作者机构： [Wu, Jing; Zhang, Shaopeng; Zhu, Wenjun; Peng, Fang; Yan, Da-zhong; Xu, Ran; Li, Xin] Wuhan Polytech Univ, Coll Biol & Pharmaceut Engn, Wuhan 430023, Hubei, Peoples R China.;[Xu, Xiaowen] Hubei Acad Forestry, Wuhan 430075, Hubei, Peoples R China.;[Wei, Wei; Chen, Weidong; Wei, W; Chen, WD] Washington State Univ, USDA ARS, Dept Plant Pathol, Pullman, WA 99164 USA.

通讯机构： [Wei, W; Chen, WD] W;Washington State Univ, USDA ARS, Dept Plant Pathol, Pullman, WA 99164 USA.

关键词： ChCAP;Colletotrichum higginsianum;Intracellular cAMP level;Pathogenicity;cAMP signaling pathway

摘要： Colletotrichum higginsianum causes anthracnose disease in a wide range of cruciferous crops and has been used as a model system to study plant-pathogen interactions and pathogenicity of hemibiotrophic plant pathogens. Conidiation, hyphae growth, appressorial development and appressorial penetration are significant steps during the infection process of C. higginsianum. However, the mechanisms of these important steps during infection remain incompletely understood. To further investigate the mechanisms of the plant-C. higginsianum interactions during infection progress, we characterized Cyclase-Associated Protein (ChCAP) gene. Deletion of the ChCAP gene resulted in reduction in conidiation and hyphal growth rate. The pathogenicity of DeltaChCAP mutants was significantly reduced with much smaller lesion on the infected leaves compared to that of wild type strain with typically water-soaked and dark necrotic lesions on Arabidopsis leaves. Further study demonstrated that the appressorial formation rate, turgor pressure, penetration ability and switch from biotrophic to necrotrophic phases decreased obviously in DeltaChCAP mutants, indicating that the attenuated pathogenicity of DeltaChCAP mutants was due to these defective phenotypes. In addition, the DeltaChCAP mutants sectored on PDA with abnormal, dark color, vesicle-like colony morphology and hyphae tip. Moreover, the DeltaChCAP mutants had a reduced intracellular cAMP levels and exogenous cAMP can partially rescue the defects of DeltaChCAP mutants in appressorial formation and penetration rate, but not in colony morphology, conidial shape and virulence, indicating that ChCAP is a key component in cAMP signaling pathway and likely play other roles in biology of C. higginsianum. In summary, our findings support the role of ChCAP in regulating conidiation, intracellular cAMP level, hyphal growth, appressorial formation, penetration ability and pathogenicity of this hemibiotrophic fungus.

语种： 英文

作者： Yan, Da-Zhong*;Gan, Ya-Ting;Zhou, Hui;Liu, Jun;Li, Xin

期刊： Current Microbiology,2018年75(3):284-287 ISSN：0343-8651

通讯作者： Yan, Da-Zhong

作者机构： [Zhou, Hui; Gan, Ya-Ting; Liu, Jun; Yan, Da-Zhong; Li, Xin] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, 68 Xuefu South Rd, Wuhan 430023, Hubei, Peoples R China.

通讯机构： [Yan, Da-Zhong] W;Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, 68 Xuefu South Rd, Wuhan 430023, Hubei, Peoples R China.

摘要： Acinetobacter sp. YT-02, a Gram-negative bacterium isolated from the activated sludge from a sodium N-cyclohexylsulfamate production plant, has the ability to degrade cyclohexylamine. It was classified as a member of Acinetobacter sp., a Gram-negative bacterium, sharing a 16S rRNA gene sequence identity of 99% with Acinetobacter guangdongensis strain 1NM-4. It could degrade 10 mmol/L cyclohexylamine within 22 h. Based on the identified metabolite, the metabolic pathway of cyclohexylamine could be postulated as it was degraded via cyclohexanone. Draft genome sequence of this strain (2,993, 647 bp of chromosome length) is presented here. We further identified the genes encoding the enzymes involved in cyclohexylamine oxidation to cyclohexanone and the subsequent downstream metabolic pathway of cyclohexanone oxidation. Strain YT-02 has the potentiality to be applied in the treatment of the pollutant cyclohexylamine, and it could also be treated as a research material to study the degradation mechanism of cyclohexylamine.

语种： 英文

作者： Zhou, Hui;Han, Zheng-gang;Fang, Ti;Chen, Yuan-yuan;Ning, Shang-bo;...

期刊： Frontiers in Microbiology,2018年9(NOV):2848 ISSN：1664-302X

通讯作者： Yan, Da-zhong

作者机构： [Han, Zheng-gang; Chen, Yuan-yuan; Ning, Shang-bo; Yan, Da-zhong; Zhou, Hui; Gan, Ya-ting] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan, Hubei, Peoples R China.;[Fang, Ti] Chinese Acad Sci, Wuhan Inst Virol, State Key Lab Virol, Wuhan, Hubei, Peoples R China.

通讯机构： [Yan, Da-zhong] W;Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan, Hubei, Peoples R China.

关键词： Acinetobacter sp. YT-02;biodegradation;cyclohexanone;cyclohexylamine;cyclohexylamine oxidase

摘要： Cyclohexylamine (CHAM) is widely used in various industries, but it is harmful to human beings and the environment. Acinetobacter sp. YT-02 can degrade CHAM via cyclohexanone as an intermediate. In this study, the cyclohexylamine oxidase (CHAO) gene from Acinetobacter sp. YT-02 was cloned. Amino acid sequence alignment indicated that the cyclohexylamine oxidase (CHAOYT-02) was 48% identical to its homolog from Brevibacterium oxydans IH-35A (CHAOIH-35). The enzyme was expressed in Escherichia coli BL21 (DE3), and purified to apparent homogeneity by Ni-affinity chromatography. The purified enzyme was proposed to be a dimer of molecular mass of approximately 91 kDa. The enzyme exhibited its maximum activity at 50°C and at pH 7.0. The enzyme was thermolabile as demonstrated by loss of important percentage of its maximal activity after 30 min incubation at 50 deg;C. Metal ions Mg2+, Co2+, and K+ had certain inhibitory effect on the enzyme activity. The kinetic parameters Km and Vmax were 0.25 ± 0.02 mM and 4.3 ± 0.083 μM min-1, respectively. The biochemical properties, substrate specificities, and three-dimensional structures of CHAOYT-02 and CHAOIH-35 were compared. Our results are helpful to elucidate the mechanism of microbial degradation of CHAM in the strain YT-02. In addition, CHAOYT-02, as a potential biocatalyst, is promising in controlling CHAM pollution and deracemization of chiral amines. © 2007 - 2018 Frontiers Media S.A.

语种： 英文

作者： Yao, Xiangyang;Lu, Binyu;Lu, Chaotian;Bai, Qin;Yan, Dazhong;...

期刊： CELL JOURNAL,2017年19(3):512-519 ISSN：2228-5806

通讯作者： Xu, Hui

作者机构： [Yao, Xiangyang; Bai, Qin; Xu, Hui; Lu, Chaotian] Bengbu Univ, Dept Biol & Food Engn, 1866 Caoshan Rd, Bengbu 233030, Peoples R China.;[Lu, Binyu] Fudan Univ, Sch Pharm, Shanghai, Peoples R China.;[Yan, Dazhong] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan, Hubei, Peoples R China.

通讯机构： [Xu, Hui] B;Bengbu Univ, Dept Biol & Food Engn, 1866 Caoshan Rd, Bengbu 233030, Peoples R China.

关键词： Apoptosis;HeLa Cells;Mitochondria;Taraxerol

摘要： Objective: Taraxerol acetate has potent anti-cancer effects via the induction of apoptosis, autophagy, cell cycle arrest, and inhibition of cell migration. However, whether taraxerol induced apoptosis and its underlying mechanisms of action is not clear. In the present study, we assess the effects of taraxerol on the mitochondrial apoptotic pathway and determine the release of cytochrome c to the cytosol and activation of caspases. Materials and Methods: In this experimental study, we mainly investigated the effect of taraxerol on HeLa cells. We tested cell viability by the MTT assay and morphologic changes, analyzed apoptosis by DAPI staining and flow cytometry. We also determined reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) using a Microplate Reader. In addition, the apoptotic proteins were tested by Western blot. Results: Taraxerol enhanced ROS levels and attenuated the MMP (Δψm) in HeLa cells. Taraxerol induced apoptosis mainly via the mitochondrial pathway including the release of cytochrome c to the cytosol and activation of caspases 9 and 3, and anti-poly (ADP-ribose) polymerase (PARP). Taraxerol could induce the down-regulation of the anti-apoptotic protein Bcl-2 and up-regulation of pro-apoptotic protein Bax. It suppressed the PI3K/ Akt signaling pathway. Conclusion: These results demonstrated that taraxerol induced cell apoptosis through a mitochondria-mediated pathway in HeLa cells. Thus, taraxerol might be a potential anti-cervical cancer candidate. Copyright © 2017 Powered By Niyasan Co.

语种： 英文

作者： Yao, Xiangyang*;Lu, Binyu;Lu, Chaotian;Bai, Qin;Yan, Dazhong;...

期刊： FOOD & FUNCTION,2017年8(1):132-141 ISSN：2042-6496

通讯作者： Yao, Xiangyang

作者机构： [Yao, Xiangyang; Bai, Qin; Xu, Hui; Lu, Chaotian; Wu, Yanli; Hong, Zibing] Bengbu Univ, Dept Biol & Food Engn, Bengbu, Peoples R China.;[Lu, Binyu] Fudan Univ, Sch Pharm, Shanghai, Peoples R China.;[Yan, Dazhong] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan, Peoples R China.

通讯机构： [Yao, Xiangyang] B;Bengbu Univ, Dept Biol & Food Engn, Bengbu, Peoples R China.

摘要： The aim of the present study was to examine the anti-inflammatory effect of solanesol and to elucidate the underlying mechanisms. Heme oxygenase-1 (HO-1) plays an important role in cytoprotection against oxidative stress and inflammation. Solanesol induced HO-1 expression both at the level of mRNA and proteins, resulting in increased HO-1 activity. Solanesol treatment enhanced the level of the phosphorylated form, nuclear translocation, ARE-binding, and transcriptional activity of Nrf2. p38 and Akt contributed to ARE-driven HO-1 expression. Solanesol activated both p38 and Akt, and treatments with SB203580 (a p38 kinase inhibitor), LY294002 (an Akt inhibitor), specific p38 siRNA and Akt siRNA suppressed the solanesol-induced activation of Nrf2, resulting in a decrease in HO-1 expression. Solanesol also elevated the autophagic protein LC3B-II level. SnPP (a HO-1 inhibitor) and HO-1 siRNA markedly abolished the anti-inflammatory effect of solanesol against LPS-induced cell damage. Likewise, SB203580, LY294002, 3-MA and Baf-A1 inhibited the solanesol-induced anti-inflammatory effect. These studies demonstrate that solanesol attenuates inflammation by HO-1 induction via p38 and Akt signaling. Thus, it is quite plausible that HO-1 induction by solanesol could trigger anti-inflammatory pathways including limiting LPS-stimulated cytokine production through autophagic signaling via p38 and Akt. © 2017 The Royal Society of Chemistry.

语种： 英文

作者： Yan, Da-Zhong*;Li, Xin;Li, Cun-Zhi;Mao, Ling-Qi;Chi, Xiang-Qun;...

期刊： Journal of Biotechnology,2017年251:166-173 ISSN：0168-1656

通讯作者： Yan, Da-Zhong

作者机构： [Li, Cun-Zhi; Mao, Ling-Qi; Yan, Da-Zhong; Li, Xin] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan 430023, Peoples R China.;[Chi, Xiang-Qun; Zhou, Ning-Yi] Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China.;[Zhou, Ning-Yi] Shanghai Jiao Tong Univ, State Key Lab Microbial Metab, Shanghai 200240, Peoples R China.;[Zhou, Ning-Yi] Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai 200240, Peoples R China.;[Liu, Dong-Yan] Shanghai Normal Univ, Coll Life & Environm Sci, Shanghai 200234, Peoples R China.

通讯机构： [Yan, Da-Zhong] W;Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan 430023, Peoples R China.

关键词： Bacteria;Carbon;Encoding (symbols);Genes;Metabolism;Polymerase chain reaction;RNA;Signal encoding;Transcription;Bioinformatics analysis;Catabolism;Cyclohexanone monooxygenase;Cyclohexanones;Cyclohexylamine;Gram-negative bacteria;Pseudomonas plecoglossicida;Reverse transcription PCR;Biodegradation;adipic acid;amine oxidase (flavin containing);cyclohexane;cyclohexanol;cyclohexanone;cyclohexylamine;cyclohexylamine oxidase;oxidoreductase;plasmid DNA;unclassified drug;cyclohexylamine derivative;Article;bacterial gene;bacterial genome;bacterium culture;carbon source;chaA gene;controlled study;gene cluster;gene identification;genome analysis;microbial degradation;nonhuman;nucleotide sequence;operon;priority journal;Pseudomonas;Pseudomonas plecoglossicida NyZ12;reverse transcription polymerase chain reaction;bacterial gene;bioremediation;genetics;metabolism;Pseudomonas;Biodegradation, Environmental;Cyclohexylamines;Genes, Bacterial;Genome, Bacterial;Oxidoreductases;Pseudomonas

摘要： The Gram-negative strain of Pseudomonas plecoglossicida NyZ12 isolated from soil has the ability to degrade cyclohexylamine (CHAM). The genes encoding CHAM degradation by gram-negative bacteria, however, have not been reported previously. In this study, ORFs predicted to encode CHAM degradation by NyZ12 were identified by bioinformatics analysis. Differential expression of the proposed ORFs was analyzed via RNA-seq and quantitative reverse transcription-PCR (qRT-PCR), using RNA extracted from NyZ12 cultured with or without CHAM addition. One CHAM-inducible ORF, RK21_02867 predicted to encode a cyclohexanone monooxygenase (ChnB) was disrupted, as were five ORFs, RK21_00425, RK21_02631, RK21_04207, RK21_04637 and RK21_05539, that had weak homology to the only known cyclohexylamine oxidase (CHAO encoded by chaA) found in Brevibacterium oxydans IH-35A. We also found that a tandem array of five ORFs (RK21_02866-02870) shared homology with those in an operon responsible for oxidation of cyclohexanone to adipic acid, although the ORFs in strain NyZ12 were arranged in a different order with previously found in cyclohexane, cyclohexanol or cyclohexanone degradation strains. The ORFs in this cluster were all up-regulated when CHAM was supplied as the sole carbon source. When one of these five genes, RK21_02867 encoding cyclohexanone (CHnone) monooxygenase, was knocked out, NyZ12 could not grow on CHAM, but it accumulated equimolar amounts of CHnone. Our results show that strain NyZ12 metabolized CHAM directly to CHnone which was then further metabolized to adipate. Despite clearly identifying genes encoding the steps for metabolism of CHAM metabolites, not every one of the putative chaAs was differentially expressed in the presence of CHAM and deletion of each one individually did not completely eliminate the capacity of NyZ12 to degrade CHAM, though it did reduce its growth in several instances. Our results suggest that there is genetic redundancy encoding the initial step in the oxidation of CHAM to CHnone in NyZ12 and that its CHAOs differ considerably from the ChaA, originally described in Brevibacterium oxydans IH-35A. © 2017 Elsevier B.V.

语种： 英文

作者： Li, Xin;Li, Cun-Zhi;Mao, Ling-Qi;Yan, Da-Zhong*;Zhou, Ning-Yi

期刊： Journal of Biotechnology,2015年199:29-30 ISSN：0168-1656

通讯作者： Yan, Da-Zhong

作者机构： [Li, Cun-Zhi; Mao, Ling-Qi; Li, Xin; Yan, Da-Zhong] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan 430023, Peoples R China.;[Zhou, Ning-Yi] Shanghai Jiao Tong Univ, State Key Lab Microbial Metab, Shanghai 200240, Peoples R China.;[Zhou, Ning-Yi] Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai 200240, Peoples R China.;[Zhou, Ning-Yi] Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China.

通讯机构： [Yan, Da-Zhong] W;Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan 430023, Peoples R China.

关键词： Bacteria;Genetic engineering;Oxidation;Complete genomes;Cyclohexanones;Cyclohexylamine;Gene clusters;Genome sequences;Gram-negative bacteria;Metabolic pathways;Pseudomonas plecoglossicida;Genes;adipic acid;cyclohexylamine;ribosome RNA;transfer RNA;cyclohexylamine derivative;Article;bacterial chromosome;bacterial genome;bacterial strain;biodegradation;gene cluster;gene sequence;Gram negative bacterium;nonhuman;nucleotide sequence;oxidation;priority journal;Pseudomonas;Pseudomonas plecoglossicida NyZ12;soil microflora;bacterial genome;genetics;metabolism;molecular genetics;multigene family;Pseudomonas;Negibacteria;Pseudomonas plecoglossicida;Cyclohexylamines;Genome, Bacterial;Molecular Sequence Data;Multigene Family;Pseudomonas

摘要： Pseudomonas plecoglossicida NyZ12 (CCTCC AB 2015057), a Gram-negative bacterium isolated from soil, has the ability to degrade cyclohexylamine. The complete genome sequence of this strain (6,233,254. bp of chromosome length) is presented, with information about the genes of characteristic enzymes responsible for cyclohexylamine oxidation to cyclohexanone and the integrated gene cluster for the metabolic pathway of cyclohexanone oxidation to adipate. © 2015 Elsevier B.V.

语种： 英文

作者： Liu, Jun;Yan, Da-zhong*;Zhao, Sheng-jun

期刊： Journal of the Science of Food and Agriculture,2015年95(13):2646-2651 ISSN：0022-5142

通讯作者： Yan, Da-zhong

作者机构： [Yan, Da-zhong; Liu, Jun] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan 430023, Peoples R China.;[Zhao, Sheng-jun] Wuhan Polytech Univ, Sch Anim Sci & Nutr Engn, Wuhan 430023, Peoples R China.

通讯机构： [Yan, Da-zhong] W;Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan 430023, Peoples R China.

关键词： fungal DNA;ribosome DNA;sweetening agent;vegetable protein;Africa;chemistry;food handling;fruit;gene expression;genetic transformation;genetics;human;Menispermaceae;metabolism;plant gene;Saccharomyces cerevisiae;Africa, Western;DNA, Fungal;DNA, Ribosomal;Food Technology;Fruit;Gene Expression;Genes, Plant;Humans;Menispermaceae;Plant Proteins;Saccharomyces cerevisiae;Sweetening Agents;Transformation, Genetic

摘要： BACKGROUND: Genetically modified (GM) foods have caused much controversy. Construction of a food-grade delivery system is a desirable technique with presumptive impact on industrial applications from the perspective of bio-safety. The aim of this study was to construct a food-grade delivery system for Saccharomyces cerevisiae and to study the expression of monellin from the berries of the West African forest plant Dioscoreophyllum cumminsii in this system. RESULTS: A food-grade system for S. cerevisiae was constructed based on ribosomal DNA (rDNA)-mediated homologous recombination to enable high-copy-number integration of the expression cassette inserted into the rDNA locus. A copper resistance gene (CUP1) was used as the selection marker for yeast transformation. Because variants of transformants containing different copy numbers at the CUP1 locus can be readily selected after growth in the presence of elevated copper levels, we suggest that this system would prove useful in the generation of tandemly iterated gene clusters. Using this food-grade system, a single-chain monellin gene was heterologously expressed. The yield of monellin reached a maximum of 675 mg L-1. CONCLUSION: This system harbors exclusively S. cerevisiae DNA with no antibiotic resistance genes, and it should therefore be appropriate for safe use in the food industry. Monellin was shown to be expressed in this food-grade delivery system. To our knowledge, this is the first report so far on expression of monellin in a food-grade expression system in S. cerevisiae. © 2015 Society of Chemical Industry.

语种： 英文

作者： Yao, Xiangyang*;Bai, Qin;Yan, Dazhong;Li, Guilan;Lu, Chaotian;...

期刊： Toxicology in Vitro,2015年29(3):600-608 ISSN：0887-2333

通讯作者： Yao, Xiangyang

作者机构： [Yao, Xiangyang; Bai, Qin; Xu, Hui; Lu, Chaotian] Bengbu Coll, Dept Biol & Food Engn, Bengbu 233030, Peoples R China.;[Yan, Dazhong] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan, Peoples R China.;[Li, Guilan] Shanxi Agr Univ, Coll Anim Sci & Vet Med, Taigu, Peoples R China.;[Yao, Xiangyang] Bengbu Coll, Dept Biol & Food Engn, 1866 Caoshan Rd, Bengbu 233030, Peoples R China.

通讯机构： [Yao, Xiangyang] B;Bengbu Coll, Dept Biol & Food Engn, 1866 Caoshan Rd, Bengbu 233030, Peoples R China.

关键词： Solanesol;HO-1;Hsp70;Nrf2;HSF1

摘要： In present study, we showed that the mRNA and protein levels of HO-1 and Hsp70 in solanesol-treated L02 cells were significantly increased. The induction of the HO-1 by solanesol is majorly achieved via enhancing the nuclear translocation and transactivity of Nrf2 through enhancement of Hsp90-Keap1 interaction, while solanesol-elevated Hsp70 is related with promoting the nuclear translocation of HSF1 through the involvement of chaperones interaction. Furthermore, the induction of HO-1 and Hsp70 by solanesol could protect against ethanol-induced liver injury, including significantly suppressing the elevation of the activities of LDH and AST, attenuating ethanol-induced increase of the MDA, ROS level and decrease of the GSH level. Moreover, solanesol also suppressed ethanol-induced apoptosis of L02 cells by inhibition of nuclear morphological damage, procaspase 3 and cleavage of caspase 3 and PARP, suggesting solanesol may be beneficial against ALD. Solanesol also promoted tBHQ-mediated protective effects. However, treatment cells with SnPP or PES markedly abrogated the protective effects of solanesol on ethanol-induced cell injury. These results strongly suggested that solanesol could protect ethanol-induced L02 cell damage, which might be attributed to the activation of HO-1 and Hsp70. (c) 2015 Elsevier Ltd. All rights reserved.

语种： 英文

作者： Yan, Da-Zhong*;Mao, Ling-Qi;Li, Cun-Zhi;Liu, Jun

期刊： World Journal of Microbiology and Biotechnology,2015年31(2):371-377 ISSN：0959-3993

通讯作者： Yan, Da-Zhong

作者机构： [Li, Cun-Zhi; Mao, Ling-Qi; Liu, Jun; Yan, Da-Zhong] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, 68 Xuefu South Rd, Wuhan 430023, Hubei, Peoples R China.

通讯机构： [Yan, Da-Zhong] W;Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, 68 Xuefu South Rd, Wuhan 430023, Hubei, Peoples R China.

关键词： Biotransformation;Cytochrome P-450cam;Hexachlorobenzene;Pentachlorophenol;Sphingobium chlorophenolicum ATCC 39723

摘要： A consortium comprised of an engineered Escherichia coli DH5α and a natural pentachlorophenol (PCP) degrader, Sphingobium chlorophenolicum ATCC 39723, was assembled for degradation of hexachlorobenzene (HCB), a persistent organic pollutant. The engineered E. coli strain, harbouring a gene cassette (camA + camB + camC) that encodes the F87W/Y96F/L244A/V247L mutant of cytochrome P-450cam (CYP101), oxidised HCB to PCP. The resulting PCP was then further completely degraded by ATCC 39723. The results showed that almost 40 % of 4 μM HCB was degraded by the consortium at a rate of 0.033 nmol/mg (dry weight)/h over 24 h, accompanied by transient accumulation and immediate consumption of the intermediate PCP, detected by gas chromatography. In contrast, in the consortium comprised of Pseudomonas putida PaW340 harbouring camA + camB + camC and ATCC 39723, PCP accumulated in PaW340 cells but could not be further degraded, which may be due to a permeability barrier of Pseudomonas PaW340 for PCP transportation. The strategy of bacterial co-culture may provide an alternative approach for the bioremediation of HCB contamination.

语种： 英文

作者： Hongmei Chen;Liping Zhang;Xianzhong Cheng;Siqing Cheng;Dazhong Yan

期刊： Asian Journal of Chemistry,2014年26(19):6579-6582 ISSN：0970-7077

通讯作者： Chen, H.

作者机构： [Cheng X.; Cheng S.; Chen H.; Zhang L.] School of Chemical and Environmental Engineering, Wuhan Polytechnic University, Wuhan, Hubei Province 430023, China;[Yan D.] School of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan, Hubei Province 430023, China

通讯机构： [Chen, H.] S;School of Chemical and Environmental Engineering, China

关键词： Adsorption;Bamboo leaves biomass;Isotherm;Kinetics;Malachite green

摘要： The native bamboo leaves biomass was investigated systematically as biosorbent for adsorption of malachite green from aqueous solution. The results show that the rapid malachite green absorption in aqueous solution by bamboo leaves biomass at pH 6 at ambient temperature observes the pseudo-second order reaction kinetics. The absorption isothermal behavior could be dissected well by Langmuir absorption isothermal, indicative of malachite green adsorbed in homogeneous surface of bamboo leaves biomass. The monolayer adsorption process for malachite green adsorption by bamboo leaves biomass and adsorption of each malachite green molecule with equal activation energy. This might be attributed to the unique morphology and composition of bamboo leaves biomass. © 2014, Chemical Publishing Co. All rights reserved.

语种： 英文

作者： Yan, Dazhong;Kang, Jianxiong;Liu, Dong-Qi*

期刊： ANNALS OF MICROBIOLOGY,2009年59(4):789-800 ISSN：1590-4261

通讯作者： Liu, Dong-Qi

作者机构： [Yan, Dazhong] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan 430023, Peoples R China.;[Liu, Dong-Qi; Kang, Jianxiong] Huazhong Univ Sci & Technol, Sch Environm Sci & Engn, Ctr Microbiol Engn, Wuhan 430074, Peoples R China.;[Liu, Dong-Qi] Huazhong Univ Sci & Technol, Sch Environm Sci & Engn, Ctr Microbiol Engn, 1037 Luoyue Rd, Wuhan 430074, Peoples R China.

通讯机构： [Liu, Dong-Qi] H;Huazhong Univ Sci & Technol, Sch Environm Sci & Engn, Ctr Microbiol Engn, 1037 Luoyue Rd, Wuhan 430074, Peoples R China.

关键词： genomic analysis;aromatic catabolic pathways;Silicibacter pomeroyi DSS-3

摘要： Genomic analysis of the catabolic potentialities of Silicibacter pomeroyi DSS-3 against a wide range of natural aromatic compounds and sequence comparisons with the entire genome of this microorganism predicted the existence of at least seven main pathways for the conversion of the aromatic compounds to the intermediates which enter into TCA cycle, that is, the catechol (cat I and cat II genes) and protocatechuate (pca genes) branches of the β-ketoadipate pathway, the phenylacetate pathway (paa genes), the gentisate pathway (gtd genes), the homogentisate pathway (hmg/hppD genes), as well as the homoprotocatechuate pathway (hpc genes). Furthermore, the genes encoding those enzymes involved in the peripheral pathways leading to the β-ketoadipate central pathway were also mapped, i.e., 4-hydroxybenzoate (pob), benzoate (ben), quinate (qui), phenylpropenoid compounds (fcs, ech, vdh, cal, van, acd and act), tyrosine (hpp) and n-phenylalkanoic acids (fad). Evidences showed that S. pomeroyi DSS-3 have versatile abilities to the catabolism of aromatic compounds either in anaerobic or in aerobic pathway, suggesting such a strain might be a model of heuristic value for the study of the genomic organization, the evolution of genes, as well as the catalytic or transcriptional mechanisms of enzymes for aromatic degradation in marine bacteria. Further, it would provide new insights into the biodegradation of aromatic compounds in marine bacteria and marine environments.

语种： 英文

作者： 杨书慧;赵胜军;刘军;闫达中;易丹

期刊： 中国生物工程杂志,2008年28(3):53-58 ISSN：1671-8135

作者机构： 武汉工业学院动物营养与饲料科学湖北省重点实验室,武汉,430023;武汉工业学院生物与制药工程系,武汉,430023;[刘军; 杨书慧; 闫达中; 赵胜军; 易丹] 武汉工业学院

关键词： 甜蛋白Monellin;乳糖;诱导表达;优化

摘要： 根据已报道的单链Monellin甜蛋白的氨基酸序列,按大肠杆菌基因偏爱密码子,设计和人工合成了单链monellin基因。将单链monellin基因克隆到大肠杆菌表达载体pET-28a中,构建了重组表达载体pET28a-mon,转化大肠杆菌BL21（DE3）,得到表达Monellin的大肠杆菌工程菌株。借助SDS-PAGE分析方法,研究了乳糖代替IPTG诱导大肠杆菌表达甜蛋白Monellin。通过对乳糖作为诱导剂表达条件进行优化,Monellin的表达量可占细胞总蛋白的33.09%,与IPTG诱导表达量接近。为乳糖作为诱导剂应用于重组大肠杆菌生产甜蛋白Monellin提供了参考依据。

语种： 中文