Genome sequence of lung pathogenic Escherichia coli O78, a chimeric strain isolated from pneumonia forest musk deer

Qing Tian,Xin Zhou,Jianguo Cheng,Yan Luo,Lei Dai,Wei Zhao,Wuyou Wang 한국유전학회 2017 Genes & Genomics Vol.39 No.7

Pneumonia is one of the major diseases of forest musk deer (FMD) that affects the survival of musk deer breeding farms. Lung pathogenic Escherichia coli (LPEC) strains were found to be one of the principal bacterial pathogens, and O78 was found to be the dominant serotype and the most poisonous, thus it was selected as further study. Here we have finished the LPEC O78 genome sequence, genomic comparative analysis and genome annotation. In the genome of LPEC O78, genes encoded major virulence factors (VFs) of urinary tract infection (UTI) and neonatal meningitis-causing E. coli (NMEC) were detected. The phylogenetic analysis indicated that LPEC O78 belongs to the D group of E. coli and clustered with human uropathogenic E. coli (UPEC) UMNO26. Orthologous analysis showed that LPEC O78 was close to UMNO26 in evolutionary relationships, the results were consistent with phylogenetic analysis. Additionally, analysis of the specific genes using COG, GO and Swiss-Prot databases revealed specific functions in the LPEC O78, some of these differences might reflect the pathogenicity of LPEC O78. It was the first time that LPEC from FMD was sequenced, through the virulence gene analysis, an example of a genome of chimeric pathogenic properties was found. The results were in favor of the possibility that strain of animal origin LPEC O78 was dangerous for public health and consequently constitute a zoonotic risk. Moreover, the functions study of specific genes will facilitate understand the pathogenicity of LPEC O78, and aid in the development of control measures.

Biotransformation and detoxification of aflatoxin B₁ by extracellular extract of Cladosporium uredinicola

Shao, Shuai,Cai, Jun,Du, Xin,Wang, ChangGao,Lin, JianGuo,Dai, Jun 한국식품과학회 2016 Food Science and Biotechnology Vol.25 No.6

Aflatoxin contamination of food and grain poses a serious economic and health problem globally. Aflatoxin $B_1$ ($AFB_1$) is extremely mutagenic and toxic as well as a potent carcinogen to both humans and livestock. In this study, the degradation of $AFB_1$ by extracellular extract of Cladosporium uredinicola was examined using high-performance liquid chromatography (HPLC), thin-layer chromatography (TLC), and liquid chromatography mass spectrometry. Within 24 h of incubation, $AFB_1$ was efficiently eliminated by the culture supernatant of C. uredinicola ($84.5{\pm}5.7%$) at $37^{\circ}C$; the elimination was proven to be enzymatic, and the enzyme was thermostable. The biotransformation products of $AFB_1$ detected by HPLC and TLC were proven to be the same compound. Analysis with LCMS showed that $AFB_1$ was bio-transformed to a structurally different compound ($m/z=365[M+Na]^+$), which is first reported. The cytotoxicity study to HeLa cells indicated that culture supernatant-treated $AFB_1$ is less toxic as compared with $AFB_1$.

Biotransformation and detoxification of aflatoxin B1 by extracellular extract of Cladosporium uredinicola

Shuai Shao,Jun Cai,Xin Du,Changgao Wang,Jianguo Lin,Jun Dai 한국식품과학회 2016 Food Science and Biotechnology Vol.25 No.6

Aflatoxin contamination of food and grain poses a serious economic and health problem globally. Aflatoxin B1 (AFB1) is extremely mutagenic and toxic as well as a potent carcinogen to both humans and livestock. In this study, the degradation of AFB1 by extracellular extract of Cladosporium uredinicola was examined using high-performance liquid chromatography (HPLC), thin-layer chromatography (TLC), and liquid chromatography mass spectrometry. Within 24 h of incubation, AFB1 was efficiently eliminated by the culture supernatant of C. uredinicola (84.5±5.7%) at 37oC; the elimination was proven to be enzymatic, and the enzyme was thermostable. The biotransformation products of AFB1 detected by HPLC and TLC were proven to be the same compound. Analysis with LCMS showed that AFB1 was bio-transformed to a structurally different compound (m/z=365 [M+Na]+), which is first reported. The cytotoxicity study to HeLa cells indicated that culture supernatant-treated AFB1 is less toxic as compared with AFB1.

Protein target identifi cation of ginsenosides in skeletal muscle tissues: discovery of natural smallmolecule activators of muscle-type creatine kinase

Feiyan Chen,Kexuan Zhu,Lin Chen,Liufeng Ouyang,Cuihua Chen,Ling Gu,Yucui Jiang,Zhongli Wang,Zixuan Lin,Qiang Zhang,Xiao Shao,Jianguo Dai,Yunan Zhao 고려인삼학회 2020 Journal of Ginseng Research Vol.44 No.3

Background: Ginseng effectively reduces fatigue in both animal models and clinical trials. However, themechanism of action is not completely understood, and its molecular targets remain largely unknown. Methods: By screening for proteins that interact with the primary components of ginseng (ginsenosides)in an affinity chromatography assay, we have identified muscle-type creatine kinase (CK-MM) as a potentialtarget in skeletal muscle tissues. Results: Biolayer interferometry analysis showed that ginsenoside metabolites, instead of parent ginsenosides,had direct interaction with recombinant human CK-MM. Subsequently, 20(S)-protopanaxadiol(PPD), which is a ginsenoside metabolite and displayed the strongest interaction with CK-MM in thestudy, was selected as a representative to confirm direct binding and its biological importance. Biolayerinterferometry kinetics analysis and isothermal titration calorimetry assay demonstrated that PPDspecifically bound to human CK-MM. Moreover, the mutation of key amino acids predicted by moleculardocking decreased the affinity between PPD and CK-MM. The direct binding activated CK-MM activityin vitro and in vivo, which increased the levels of tissue phosphocreatine and strengthened the functionof the creatine kinase/phosphocreatine system in skeletal muscle, thus buffering cellular ATP, delayingexercise-induced lactate accumulation, and improving exercise performance in mice. Conclusion: Our results suggest a cellular target and an initiating molecular event by which ginsengreduces fatigue. All these findings indicate PPD as a small molecular activator of CK-MM, which can helpin further developing better CK-MM activators based on the dammarane-type triterpenoid structure.

Protein target identification of ginsenosides in skeletal muscle tissues: discovery of natural small-molecule activators of muscle-type creatine kinase

Chen, Feiyan,Zhu, Kexuan,Chen, Lin,Ouyang, Liufeng,Chen, Cuihua,Gu, Ling,Jiang, Yucui,Wang, Zhongli,Lin, Zixuan,Zhang, Qiang,Shao, Xiao,Dai, Jianguo,Zhao, Yunan The Korean Society of Ginseng 2020 Journal of Ginseng Research Vol.44 No.3

Background: Ginseng effectively reduces fatigue in both animal models and clinical trials. However, the mechanism of action is not completely understood, and its molecular targets remain largely unknown. Methods: By screening for proteins that interact with the primary components of ginseng (ginsenosides) in an affinity chromatography assay, we have identified muscle-type creatine kinase (CK-MM) as a potential target in skeletal muscle tissues. Results: Biolayer interferometry analysis showed that ginsenoside metabolites, instead of parent ginsenosides, had direct interaction with recombinant human CK-MM. Subsequently, 20(S)-protopanaxadiol (PPD), which is a ginsenoside metabolite and displayed the strongest interaction with CK-MM in the study, was selected as a representative to confirm direct binding and its biological importance. Biolayer interferometry kinetics analysis and isothermal titration calorimetry assay demonstrated that PPD specifically bound to human CK-MM. Moreover, the mutation of key amino acids predicted by molecular docking decreased the affinity between PPD and CK-MM. The direct binding activated CK-MM activity in vitro and in vivo, which increased the levels of tissue phosphocreatine and strengthened the function of the creatine kinase/phosphocreatine system in skeletal muscle, thus buffering cellular ATP, delaying exercise-induced lactate accumulation, and improving exercise performance in mice. Conclusion: Our results suggest a cellular target and an initiating molecular event by which ginseng reduces fatigue. All these findings indicate PPD as a small molecular activator of CK-MM, which can help in further developing better CK-MM activators based on the dammarane-type triterpenoid structure.