宁夏地区牛源金黄色葡萄球菌β-内酰胺酶的分析及作用方式的研究

doi:10.11843/j.issn.0366-6964.2020.05.025

Abstract

Abstract: This study aimed to study the evolution and structural characteristics of β-lactamase produced by Staphylococcus aureus (SA) and its action mode in Ningxia province. Antimicrobial susceptibility test and PCR were used to detect the resistance and drug resistance genes of five β-lactams respectively; the evolution analysis and structure function prediction of β-lactamase BlaZ were made by bioinformatics; the mode of action of BlaZ was analyzed by molecular docking and dynamic simulation. The results showed that the resistance rate of Penicillins was significantly higher than Cephalosporin's; the detection rate of blaZ gene was 82.37% and the detection rate of blaZ_TEM-1 gene, which was rarely reported in SA, was 26.05%. According to evolutionary analysis, BlaZ belongs to A-type β-lactamase and contains 63 important trace residues under evolutionary pressure. Meanwhile, molecular docking showed that BlaZ is easy to combine with Penicillins, Avibatan, what's more, formed stable. The results of molecular dynamics simulation showed that the structural flexibility of BlaZ was less than that of TEM-1 and TEM-52, in which there was a difference in the flexibility of Ω-loop region (P<0.05) and it could affect the antibiotic binding. Thus, there was a positive correlation between the flexibility of the region and the binding activity; the structural stability of BlaZ was significantly different (P<0.05) when it was combined with Ampicillin and Ceftiofur, respectively. The BlaZ produced by SA in Ningxia is a A-type β-lactamase. In addition, blaZ_TEM-1 gene was detected from those strains. The bind-ing activity of BlaZ to Penicillins and Avibactam is higher than that of Cephalosporin, and the structural flexibility of Ω-loop region is an important factor affecting the binding activity.

Key words: Staphylococcus aureus, β-lactamase, action mode, bioinformatics, molecular simulation

CLC Number:

S852.611

MA Qiang, YANG Rui, WAN Jiahong, CHANG Jiawei, WEI Yanqin, WANG Guiqin. Study on β-lactamase and Its Action Mode of Staphylococcus aureus Isolated from Cows in Ningxia[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(5): 1138-1148.

References 31

[1]	CANTÓN R, LOZA E, AZNAR J, et al. Antimicrobial susceptibility trends and evolution of isolates with extended spectrum β-lactamases among Gram-negative organisms recovered during the SMART study in Spain (2011-2015)[J]. Rev Esp Quimioter, 2018, 31(2):136-145.
[2]	MALLOY A M, CAMPOS J M. Extended-spectrum beta-lactamases:a brief clinical update[J]. Pediatr Infect Dis J, 2011, 30(12):1092-1093.
[3]	RUGGIERO M, CURTO L, BRUNETTI F, et al. Impact of mutations at Arg220 and Thr237 in PER-2β-lactamase on conformation, activity, and susceptibility to inhibitors[J]. Antimicrob Agents Chemother, 2017, 61(6):e02193-16.
[4]	DAS C K, NAIR N N. Hydrolysis of cephalexin and meropenem by New Delhi metallo-β-lactamase:the substrate protonation mechanism is drug dependent[J]. Phys Chem Chem Phys, 2017, 19(20):13111-13121.
[5]	COLLIGNON P, BEGGS J J, WALSH T R, et al. Anthropological and socioeconomic factors contributing to global antimicrobial resistance:a univariate and multivariable analysis[J]. Lancet Planet Health, 2018, 2(9):e398-e405.
[6]	MIHALEK I, REŠ I, LICHTARGE O. Evolutionary trace report_maker:a new type of service for comparative analysis of proteins[J]. Bioinformatics, 2006, 22(13):1656-1657.
[7]	WANG Y B, LI X L, JIANG L, et al. Novel mutation sites in the development of vancomycin-intermediate resistance in Staphylococcus aureus[J]. Front Microbiol, 2016, 7:2163.
[8]	WATTS J L. Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals:approved standard[M]. Third Edition. Pennsylvania:Clinical and Laboratory Standards Institute (CLSI), 2008.
[9]	STVRENBURG E, KVHN A, MACK D, et al. A novel extended-spectrum β-lactamase CTX-M-23 with a P167T substitution in the active-site omega loop associated with ceftazidime resistance[J]. J Antimicrob Chemother, 2004, 54(2):406-409.
[10]	LIU H H, WANG Y L, WANG G, et al. The prevalence of Escherichia coli strains with extended spectrum beta-lactamases isolated in China[J]. Front Microbiol, 2015, 6:335.
[11]	DONG Y Y, SHENG H H, ZENG X T, et al. Investigation of genetic diversity of the blaSHV gene and development of an oligonucleotide microarray to detect mutations in the blaSHV gene[J]. Microb Drug Resist, 2012, 18(6):539-545.
[12]	POIREL L, GEROME P, DE CHAMPS C, et al. Integron-located oxa-32 gene cassette encoding an extended-spectrum variant of OXA-2β-lactamase from Pseudomonas aeruginosa[J]. Antimicrob Agents Chemother, 2002, 46(2):566-569.
[13]	PORTO A, AYALA J, GUTKIND G, et al. A novel OXA-10-like β-lactamase is present in different Enterobacteriaceae[J]. Diagn Microbiol Infect Dis, 2010, 66(2):228-229.
[14]	AUBERT D, NAAS T, NORDMANN P. Integrase-mediated recombination of the veb1 gene cassette encoding an extended-spectrum β-lactamase[J]. PLoS One, 2012, 7(12):e51602.
[15]	PASTERÁN F, RAPOPORT M, PETRONI A, et al. Emergence of PER-2 and VEB-1a in Acinetobacter baumannii strains in the Americas[J]. Antimicrob Agents Chemother, 2006, 50(9):3222-3224.
[16]	DA SILVA J V, FERREIRA L D, ALVES L R, et al. Detection of multidrug-resistant Pseudomonas aeruginosa harboring blaGES-1 and blaGES-11 in Recife, Brazil[J]. Rev Soc Bras Med Trop, 2017, 50(6):764-768.
[17]	POND S L K, FROST S D W, MUSE S V. HyPhy:hypothesis testing using phylogenies[J]. Bioinformatics, 2005, 21(5):676-679.
[18]	SPIELMAN S J, KOSAKOVSKY POND S L. Relative evolutionary rate inference in HyPhy with LEISR[J]. PeerJ, 2018, 6:e4339.
[19]	PARRA R G, SCHAFER N P, RADUSKY L G, et al. Protein Frustratometer 2:a tool to localize energetic frustration in protein molecules, now with electrostatics[J]. Nucleic Acids Res, 2016, 44(W1):W356-W360.
[20]	TOOKE C L, HINCHLIFFE P, BRAGGINTON E C, et al. β-lactamases and β-lactamase inhibitors in the 21st century[J]. J Mol Biol, 2019, 431(18):3472-3500.
[21]	YOUSEFIPOUR M, RASOULINEJAD M, HADADI A, et al. Bacteria producing extended spectrum β-lactamases (ESBLs) in hospitalized patients:prevalence, antimicrobial resistance pattern and its main determinants[J]. Iran J Pathol, 2019, 14(1):61-67.
[22]	NOLIVOS S, CAYRON J, DEDIEU A, et al. Role of AcrAB-TolC multidrug efflux pump in drug-resistance acquisition by plasmid transfer[J]. Science, 2019, 364(6442):778-782.
[23]	VAJRAVIJAYAN S, PLETNEV S, MANI N, et al. Structural insights on starch hydrolysis by plant β-amylase and its evolutionary relationship with bacterial enzymes[J]. Int J Biol Macromol, 2018, 113:329-337.
[24]	MORGAN D H, KRISTENSEN D M, MITTELMAN D, et al. ET viewer:an application for predicting and visualizing functional sites in protein structures[J]. Bioinformatics, 2006, 22(16):2049-2050.
[25]	GE Y Y, CHI Y, MIN X Y, et al. The evolution and characterization of influenza A(H7N9) virus under the selective pressure of peramivir[J]. Virology, 2019, 536:58-67.
[26]	PALZKILL T. Structural and mechanistic basis for extended-spectrum drug-resistance mutations in altering the specificity of TEM, CTX-M, and KPC β-lactamases[J]. Front Mol Biosci, 2018, 5:16.
[27]	YI H, CHOI J M, HWANG J, et al. High adaptability of the omega loop underlies the substrate-spectrum-extension evolution of a class A β-lactamase, PenL[J]. Sci Rep, 2016, 6(1):36527.
[28]	HART K M, HO C M W, DUTTA S, et al. Modelling proteins' hidden conformations to predict antibiotic resistance[J]. Nat Commun, 2016, 7(1):12965.
[29]	PAPP-WALLACE K M, BECKA S A, TARACILA M A, et al. Exploring the role of the Ω-loop in the evolution of Ceftazidime resistance in the PenA β-lactamase from Burkholderia multivorans, an important cystic fibrosis pathogen[J]. Antimicrob Agents Chemother, 2017, 61(2):e01941-16.
[30]	PEREIRA R, RABELO V W H, SIBAJEV A, et al. Class A β-lactamases and inhibitors:in silico analysis of the binding mode and the relationship with resistance[J]. J Biotechnol, 2018, 279:37-46.
[31]	DOCQUIER J D, MANGANI S. An update on β-lactamase inhibitor discovery and development[J]. Drug Resist Updat, 2018, 36:13-29.

Study on β-lactamase and Its Action Mode of Staphylococcus aureus Isolated from Cows in Ningxia

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 31

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	ZHOU Yang, WU Weizi, CAO Weisheng, WANG Fuguang, XU Xiuqiong, ZHONG Wenxia, WU Liyang, YE Jian, LU Shousheng. A Whole Genome Sequencing Method for African Swine Fever Virus based on Nanopore Sequencing Technology was Established [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2080-2089.
[2]	HE Xiaolan, ZHAO Yankun, MENG Lu, LIU Huimin, GAO Jiaojiao, ZHENG Nan. Research Progress in Heteroresistance of Staphylococcus aureus [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1432-1445.
[3]	CAO Yuzhu, XING Yuxin, MA Chenglin, GUAN Hongbo, JIA Qihui, KANG Xiangtao, TIAN Yadong, LI Zhuanjian, LIU Xiaojun, LI Hong. Biological Characterization of Chicken FGF6 Gene and Association of Its Polymorphisms with Economic Traits [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1536-1550.
[4]	YANG Yang, YU Qian, LIU Yucheng, YANG Hua, ZHAO Zhuo, WANG Limin, ZHOU Ping, YANG Qingyong, DAI Rong. Identification and Tissue Expression Analysis of the Sheep MYL Gene Family [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1551-1564.
[5]	TIAN Rui, XU Sixiang, XIE Feng, LIU Guangjin, WANG Gang, LI Qingxia, DAI Lei, XIE Guoxin, ZHANG Qiongwen, LU Yajing, WANG Guangwen, WANG Jinxiu, ZHANG Wei. Bioinformatics Analysis of the Genome of Clostridium perfringens Isolated from Cattle [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1707-1715.
[6]	WU Zihao, CAI Yilong, TUO Haixin, CHEN Wei. Pathogenicity Analysis of a PVL⁺ ST22 Staphylococcus aureus Isolated from Equine Raw Milk [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 718-726.
[7]	DU Xiaodi, HOU Wei, SU Zhonghua, MA Qingmei, HE Xue, HUA Ruiqi, YANG Aiguo, YANG Guangyou. Bioinformatics and Expression Analysis of Ubiquitin-conjugating Enzyme Gene Family of Echinococcus granulosus [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2605-2618.
[8]	MENG Qiuchi, LU Guangyu, CHEN Dingshuang, LIN Yaqiu, WANG Ruilong, ZHONG Chaosong, WANG Yong, LIU Wei, WANG Youli, LI Yanyan, LI Zhixiong. Goats GPR35 Gene Expression Characteristics Analysis and the Effect on Subcutaneous Fat Cell Differentiation [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 4993-5007.
[9]	JIANG Nansong, JI Xing, WANG Yaxin, SUN Chengtao, WANG Yang, CHEN Hongmei, CHENG Longfei, HUANG Yu, WU Congming. Prevalence and Transduction of Prophages in Methicillin-resistant Staphylococcus aureus ST9 of Swine Origin [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 338-350.
[10]	YANG Yang, ZHOU Ziwei, ZHANG Jingyi, YANG Shuo, WANG Boyu, GE Nan, LIN Ye, HOU Xiaoming. Analysis on SP1 Gene Structure and Its Function on Milk Fat Synthesis in Holstein Dairy Cows [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(9): 2970-2981.
[11]	LU Wanqing, ZHAO Shasha, JIANG Songhong, TONG Zhizi, HUANG Danni, GUO Jianhua, WU Junwei, ZHOU Yang. Effect of Staphylococcus aureus on IFN-α Production in BV2 Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2633-2641.
[12]	CEN Xin, YANG Tingting, ZHAO Zunfu, WEN Yongping, ZHANG Huanrong. Isolation, Identification, Biological Characteristics and Genome Analysis of a Virulent Salmonella Phage [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2677-2688.
[13]	MAO Yanni, CHANG Jiawei, LI Na, WANG Xin, KANG Xinyun, MA Qiang, MA Liang, WANG Guiqin. Transcriptome Differential Expression Analysis of Staphylococcus aureus in Biofilm State and Planktonic State [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2697-2707.
[14]	GAO Dengke, ZHAO Hongcong, DONG Hao, JIN Yaping, CHEN Huatao. The Cloning, Expression Vector Construction and Function Analysis of Goat RORα Gene [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(6): 1779-1794.
[15]	HUANG Min, XIE Xiaogang, HE Qifu, CAO Xuyang, DONG Xiangchen, KANG Jian, LIU Jun, QUAN Fusheng. Cloning Analysis and Construction of Knockout Vector of Zfy Gene in Goat [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(3): 711-721.