Isolation and Characterization of Quorum-quenching Genes from Bacterium Pseudomonas sp. WX14

doi:10.16409/j.cnki.2095-039x.2018.01.013

Abstract

Abstract: In this study, ten strains with strong AHLs-degrading activity were obtained through "reporter plate" and the assay of β-galactosidase activity. Strain WX14 displayed the highest AHLs-degrading activity and the AHLs-degrading capacity was observed in bacterial cells and extracellular secretions. Quorum quenching genes hacA and hacB were identified and cloned from strain WX14, and full lengths of these two genes are 2286 bp and 2370 bp, respectively. Analysis of amino sequence similarities showed that HacA belongs to aculeacin A acylase family, and HacB belongs to penicillin G acylase family. Both HacA and HacB are members of the N-terminal nucleophile hydrolase superfamily. It was observed that Pectobacterium carotovorum Z3-3 after introduced hacA or hacB attenuated its pathogenicity in potato tubers and Chinese cabbages, indicating these two AHLs degrading enzymes might have potential to control of the QS-dependent soft rot disease. BLASTn analysis of bacterial 16S rDNA sequences showed that ten isolates were identified as Pseudarthrobacter spp. (3 isolates), Paenarthrobacter sp. (1 isolate), Pseudomonas spp. (4 isolates) and Bacillus spp. (2 isolates), respectively.

Key words: quorum quenching, acyl-homoserine lactones, acylase, N-terminal nucleophile hydrolytic enzymes

CLC Number:

S476

ZHANG Qingxia, ZHANG Ying, JI Yanyan, TONG Yunhui, JI Zhaolin. Isolation and Characterization of Quorum-quenching Genes from Bacterium Pseudomonas sp. WX14[J]. journal1, 2018, 34(1): 109-116.

References

[1] Tang K, Zhang Y, Yu M, et al. Evaluation of a new high-throughputmethod for identifying quorum quenching bacteria[J]. Scientific Report, 2013, 3(10):2935.
[2] Crépin A, Beury-Cirou A, Barbey C, et al. N-acyl homoserine lactones indiverse Pectobacterium and Dickeya plant pathogens:diversity, abundance, and involvement in virulence[J]. Sensors, 2012, 12(3):3484-3497.
[3] Cui X, Harling R. N-acyl-homoserine lactone-mediated quorum sensing blockage, a novel strategy for attenuating pathogenicity of gram-negative bacterial plant pathogens[J]. European Journal of Plant Pathology, 2005, 111(4):327-339.
[4] Schell M A. To be or not to be:how Pseudomonas solanacearum decides whether or not to express virulence genes[J]. European Journal of Plant Pathology, 1996, 102:459-469.
[5] 任争光, 林敏, 姜文君, 等. 群体感应系统对甜瓜果斑病菌MH21致病力的影响[J]. 植物病理学报, 2012, 42(6):608-619.
[6] LaSarre B, Federle M J. Exploiting quorum sensing to confuse bacterial pathogens[J]. Microbiology and Molecular Biology Reviews, 2013, 77(1):73-111.
[7] 张力群, 田涛, 梅桂英. 群体感应淬灭——防治植物细菌病害的新策略[J]. 中国生物防治, 2010, 26(3):241-247.
[8] Kalia V C, Raju S C, Purohit H J. Genomic analysis reveals versatile organisms for quorum quenching enzymes:acyl-homoserine lactone-acylase and -lactonase[J]. Open Microbiology Journal, 2011, 5(1):1-13.
[9] Dong Y H, Xu J L, Li X Z, et al. AiiA, an enzyme that inactivates the acylhomoserine lactonequorum-sensing signal and attenuates the virulence of Erwinia carotovora[J]. Proceedings of the National Academy of Sciences of the United States of America, 2000, 97(7):3526-3531.
[10] Dong Y H, Wang L H, Xu J L, et al. Quenching quorum-sensing-dependent bacterial infection by an N-acyl homoserine lactonase[J]. Nature, 2001, 411:813-817.
[11] Fan J, Qian G L, Chen T, et al. Biocontrol of bacterial soft rot of callalily by elicitor Harpin_xoo and N-acyl homoserine lactonase (AttM)[J]. World Journal of Microbiology and Biotechnology, 2010, 27(2):401-410.
[12] Christiaen S E A, Brackman G, Nelis H J, et al. Isolation and identification of quorum quenching bacteria from environmental samples[J]. Journal of Microbiological Methods, 2011, 87:213-219.
[13] Wei H L, Zhang L Q. Quorum-sensing system influences root colonization and biological control ability in Pseudomonas fluorescens 2P24[J]. Antonie van Leeuwenhoek, 2006, 89(2):267-280.
[14] Mei G Y, Yan X X, Turak A, et al. AidH, an alpha/beta-hydrolase fold family member from an Ochrobactrum sp. strain, is a novel N-acylhomoserine lactonase[J]. Applied and Environmental Microbiology, 2010, 76(15):4933-4942.
[15] Heeb S, Blumer C, Haas D. Regulatory RNAas mediator in GacA/RsmA-dependent global control of exoproduct formation in Pseudomonas fluorescens CHA0[J]. Journal of Bacteriology, 2002, 184(4):1046-1056.
[16] Marchesi J R, Sato T, Weightman A J, et al. Design and evaluation of useful bacterium-specific PCR primers that amplify genes coding for bacterial 16S rRNA[J]. Applied and Environmental Microbiology, 1998, 64(2):795-799.
[17] Tamura K, Peterson D, Peterson N, et al. MEGA5:molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods[J]. Molecular Biology Evolution, 2011, 28(10):2731-2739.
[18] Ochiai S, Yasumoto S, Morohoshi T, et al. AmiE, a novel N-acylhomoserinelactone acylase belonging to the amidase family, from the activated-sludge isolate Acinetobacter sp. strain Ooi24[J]. Applied and Environmental Microbiology, 2014, 80(22):6919-6925.
[19] Fetzner S. Quorum quenching enzymes[J]. Journal of Biotechnology, 2015, 201:2-14.
[20] Lin Y H, Xu J L, Hu J Y, et al. Acyl-homoserine lactone acylase from Ralstonia strain XJ12B represents a noveland potent class of quorum-quenching enzymes[J]. Molecular Microbiology, 2003, 47(3):849-860.
[21] Huang J J, Petersen A, Whiteley M, et al. Identification of QuiP, the product of gene PA1032, asthe second acyl-homoserine lactone acylase of Pseudomonas aeruginosa PAO1[J]. Applied and Environmental Microbiology, 2006, 72(2):1190-1197.
[22] Wahjudi M, Papaioannou E, Hendrawati O, et al. PA0305 of Pseudomonas aeruginosaisa quorum quenching acylhomoserine lactone acylase belonging to the Ntn hydrolase superfamily[J]. Microbiology, 2011, 157(7):2042-2055.
[23] Park S Y, Kang H O, Jang H S, et al. Identifcation of extracellular N-acylhomoserine lactoneacylase from a Streptomyces sp. and its application to quorum quenching[J]. Applied and Environmental Microbiology, 2005, 71(5):2632-2641.
[24] Romero M, Diggle S P, Heeb S, et al. Quorum quenching activity in Anabaena sp. PCC7120:identification of AiiC, a novel AHL-acylase[J]. FEMS Microbiology Letters, 2008, 280(1):73-80.
[25] Shepherd R W, Lindow S E. Two dissimilar N-acyl-homoserine lactone acylases ofPseudomonas syringae influence colony and biofilm morphology[J]. Applied and Environmental Microbiology, 2009, 75(1):45-53.
[26] Dong Y H, Wang L Y, Zhang L H. Quorum-quenching microbial infections:mechanisms and implications[J]. Philosophical Transactions of the Royal Society of London[J]. 2007, 362(1483):1201-1211.
[27] Molina L, Constantinescu F, Michel L, et al. Degradation of pathogen quorum-sensing molecules by soil bacteria:a preventive and curative biological control mechanism[J]. FEMS Microbiology Ecology, 2003, 45(1):71-81.
[28] Uroz S, Heinonsalo J. Degradation of N-acyl homoserine lactone quorum sensing signal molecules by forest root-associated fungi[J]. FEMS Microbiology Ecology, 2008, 65(2):271-278.
[29] Cha C, Gao P, Chen Y C, et al. Production of acyl-homoserine lactone quorum-sensing signals by gram-negative plant-associated bacteria[J]. Molecular Plant-Microbe Interactions, 1998, 11(11):1119-1129.