Bacterial pathogenesis-associated characteristics such as biofilm formation, synthesis of hydrolyzing enzymes, and toxins are regulated by Acyl Homoserine Lactones (AHLs), small peptides and diffusing signal factors (DSF). Lelliottia amnigena is gram negative bacteria and its pathogenicity is regulated by the luxR and luxI class of quorum sensing. The signaling molecules and their concentrations are essential for the virulence of the pathogenic bacterium. To suppresses the pathogenicity; the concentration of signalling molecules must be controlled or degraded. The lactonase have the ability to hydrolyze lactones of different chain length. The present study deals with a newer approach to control the pathogenesis of Lelliottia amnigena through isolation and characterization of Aiia lactonase from Bacillus cereus RC1. Aiia lactonase specific primers were used to amplify the gene, and the sequence thus obtained was submitted to the Genbank database under accession # OK643884.1. The gene was cloned in pBE-S shuttle vector and transformed in the recombinant host. The expressed and purified protein had a molecular weight of 28.00 KDa and exhibited its optimum activity at 37℃ by inhibiting the violacein pigment of the monitor strain Chromobacterium violaceum MTCC 2656. The proteinaceous nature of the purified molecule was confirmed by incubating it in the presence of proteinase K for 1 h. The activity of the pathogenesis-related protein, polygalacturonase was drastically reduced in the presence of the purified Aiia protein. The purified protein also showed a zone of inhibition when plated together with Lelliottia amnigena RCE (MZ712952.1). Searches of the Conserved Domain Database suggested that this protein belonged to the Metallo-beta-lactamase superfamily and is closely related to Aiia from B. thuringiensis serovar kurstaki. Modeling of the protein structure was done using I-TASSER; a C-score of 0.55 suggested that the model was of good quality. To be used commercially, this recombinant protein needs to be purified at an industrial scale; it can then be used to repress the growth of soft rot causing bacteria in horticultural crops during their storage period.

ASPEE Shakilam Biotech Institute Navsari Agricultural University Surat Gujarat India

Department of Biology University of Waterloo Waterloo ON N2L 3G1 Canada

Department of Geology and Soil Science Faculty of Forestry and Wood Technology Mendel University in Brno Brno Czech Republic

Department of Plant Molecular Biology and Biotechnology ASPEE College of Horticulture and Forestry Navsari Agricultural University Navsari 396450 Gujarat India

Hainan Key Laboratory for Sustainable Utilization of Tropical Bioresource College of Tropical Crops Hainan University Haikou 570228 China

Zobrazit více v PubMed

Abd-Elhafeez ESE (2018) Isolation and characterization of two bacteriophages infecting Kosakonia sacchari bacterium causing potato soft rot disease. J Bacteriol Parasitol. https://doi.org/10.4172/2155-9597.1000344 DOI

Al-Kharousi ZS, Guizani N, Al-Sadi AM et al (2016) Hiding in fresh fruits and vegetables: opportunistic pathogens may cross geographical barriers. Int J Microbiol. https://doi.org/10.1155/2016/4292417 PubMed DOI PMC

Anandan K, Vittal RR (2019) Quorum quenching activity of AiiA lactonase (KMMI17) from endophytic Bacillus thuringiensis KMCL07 on AHL- mediated pathogenic phenotype in Pseudomonas aeruginosa. Microb Pathog 132:230–242. https://doi.org/10.1016/j.micpath.2019.05.015 PubMed DOI

Bhat KA, Masood SD, Bhat NA et al (2010) Current status of post harvest soft rot in vegetables: a review. Asian J Plant Sci 9:200–208. https://doi.org/10.3923/ajps.2010.200.208 DOI

Bottomley MJ, Muraglia E, Bazzo R, Carfì A (2007) Molecular insights into quorum sensing in the human pathogen Pseudomonas aeruginosa from the structure of the virulence regulator LasR bound to its autoinducer. J Biol Chem 282:13592–13600. https://doi.org/10.1074/jbc.M700556200 PubMed DOI

Cao Y, He S, Zhou Z et al (2012) Orally administered thermostable N-acyl homoserine lactonase from Bacillus sp. strain AI96 attenuates Aeromonas hydrophila infection in zebrafish. Appl Environ Microbiol 78:1899–1908. https://doi.org/10.1128/AEM.06139-11 PubMed DOI PMC

Carlier A, Uroz S, Smadja B et al (2003) The Ti Plasmid of Agrobacterium tumefaciens harbors an attM -paralogous gene, aiiB, also encoding N -acyl homoserine lactonase activity. Appl Environ Microbiol 69:4989–4993. https://doi.org/10.1128/AEM.69.8.4989-4993.2003 PubMed DOI PMC

Chang AY, Chau VWY, Landas JA, Pang Y (2017) Preparation of calcium competent Escherichia coli and heat-shock transformation. J Exp Microbiol Immunol 1:22–25

Chen R, Zhou Z, Cao Y et al (2010) High yield expression of an AHL-lactonase from Bacillus sp. B546 in Pichia pastoris and its application to reduce Aeromonas hydrophila mortality in aquaculture. Microb Cell Fact 9:39. https://doi.org/10.1186/1475-2859-9-39 PubMed DOI PMC

Chung YS, Goeser NJ, Cai X, Jansky S (2013) The effect of long term storage on bacterial soft rot resistance in potato. Am J Potato Res 90:351–356. https://doi.org/10.1007/s12230-013-9311-6 DOI

Collmer A, Keen NT (1986) The role of pectic enzymes in plant pathogenesis. Annu Rev Phytopathol 24:383–409. https://doi.org/10.1146/annurev.py.24.090186.002123 DOI

Czajkowski R, Pérombelon MCM, van Veen JA, van der Wolf JM (2011) Control of blackleg and tuber soft rot of potato caused by Pectobacterium and Dickeya species: a review. Plant Pathol 60:999–1013. https://doi.org/10.1111/j.1365-3059.2011.02470.x DOI

Deep A, Chaudhary U, Gupta V (2011) Quorum sensing and bacterial pathogenicity: from molecules to disease. J Lab Physicians 3:004–011. https://doi.org/10.4103/0974-2727.78553 DOI

Degefu Y, Potrykus M, Golanowska M et al (2013) A new clade of Dickeya spp. plays a major role in potato blackleg outbreaks in North Finland. Ann Appl Biol 162:231–241. https://doi.org/10.1111/aab.12020 DOI

Diallo S, Crépin A, Barbey C et al (2011) Mechanisms and recent advances in biological control mediated through the potato rhizosphere. FEMS Microbiol Ecol 75:351–364. https://doi.org/10.1111/j.1574-6941.2010.01023.x PubMed DOI

Dong Y-H (2000) AiiA, an enzyme that inactivates the acylhomoserine lactone quorum-sensing signal and attenuates the virulence of Erwinia carotovora. Proc Natl Acad Sci 97:3526–3531. https://doi.org/10.1073/pnas.060023897 PubMed DOI PMC

Dong YH, Xu JL, Li XZ, Zhang LH (2000) AiiA, an enzyme that inactivates the acylhomoserine lactone quorum-sensing signal and attenuates the virulence of Erwinia carotovora. Proc Natl Acad Sci USA 97:3526–3531. https://doi.org/10.1073/pnas.97.7.3526 PubMed DOI PMC

Dong Y-H, Zhang L-H (2005) Quorum sensing and quorum-quenching enzymes. J Microbiol 43(spc1):101–109 PubMed

Easwaran N, Karthikeyan S, Sridharan B, Gothandam KM (2015) Identification and analysis of the salt tolerant property of AHL lactonase (AiiATSAWB) of Bacillus species. J Basic Microbiol 55:579–590. https://doi.org/10.1002/jobm.201400013 PubMed DOI

Fetzner S (2015) Quorum quenching enzymes. J Biotechnol 201:2–14. https://doi.org/10.1016/j.jbiotec.2014.09.001 PubMed DOI

Florez Escobar AM, Gonzalez A, Pedroza CJ et al (2014) Identification, cloning and lactonase activity of recombinant protein of N-acyl homoserine lactonase (AiiA) from Bacillus thuringiensis 147–115-16 strain. Rev Colomb Biotecnol 16:153. https://doi.org/10.15446/rev.colomb.biote.v16n1.40495 DOI

Garnier J, Osguthorpe DJRB (1978) Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J Mol Biol 120:97–120 PubMed DOI

Gasteiger E, Gattiker A, Hoogland C et al (2003) ExPASy: the proteomics server for in-depth protein knowledge and analysis. Nucleic Acids Res 31:3784–3788. https://doi.org/10.1093/nar/gkg563 PubMed DOI PMC

Grandclément C, Tannières M, Moréra S et al (2015) Quorum quenching: role in nature and applied developments. FEMS Microbiol Rev 40:86–116. https://doi.org/10.1093/femsre/fuv038 PubMed DOI

Guruprasad K, Reddy BV, Pandit MW (1990) Correlation between stability of a protein and its dipeptide composition: a novel approach for predicting in vivo stability of a protein from its primary sequence. Protein Eng 4:155–161. https://doi.org/10.1093/protein/4.2.155 PubMed DOI

Hadizadeh I, Peivastegan B, Hannukkala A et al (2019) Biological control of potato soft rot caused by Dickeya solani and the survival of bacterial antagonists under cold storage conditions. Plant Pathol 68:297–311. https://doi.org/10.1111/ppa.12956 DOI

Hanano A, Harba M, Al-Ali M, Ammouneh H (2014) Silencing of Erwinia amylovora sy69 AHL-quorum sensing by a Bacillus simplex AHL-inducible aiiA gene encoding a zinc-dependent N-acyl-homoserine lactonase. Plant Pathol 63:773–783. https://doi.org/10.1111/ppa.12142 DOI

Ikai A (1980) Thermostability and aliphatic index of globular proteins. J Biochem 88:1895–1898 PubMed

Kabeil SS, Lashin SM, El-Masry MH et al (2008) Potato brown rot disease in Egypt: current status and prospects. J Agric Environ Sci 4:44–54

Kachhadia R, Kapadia C, Singh S et al (2022) Quorum sensing inhibitory and quenching activity of Bacillus cereus RC1 extracts on soft rot-causing Bacteria Lelliottia amnigena. ACS Omega. https://doi.org/10.1021/acsomega.2c02202 PubMed DOI PMC

Kapadia C, Alhazmi A, Patel N et al (2021) Nanoparticles combined with cefixime as an effective synergistic strategy against Salmonella enterica typhi. Saudi J Biol Sci 28:4164–4172. https://doi.org/10.1016/j.sjbs.2021.05.032 PubMed DOI PMC

Kapadia C, Kachhdia R, Singh S et al (2022) Pseudomonas aeruginosa inhibits quorum-sensing mechanisms of soft rot pathogen Lelliottia amnigena RCE to regulate its virulence factors and biofilm formation. Front Microbiol. https://doi.org/10.3389/fmicb.2022.977669 PubMed DOI PMC

Kaur H, Garg H (2014) Pesticides: environmental impacts and management strategies. In: Pesticides-toxic aspects. InTech, Rijeka

Khoiri S, Damayanti TA (2021) Characterization of acyl-homoserine lactonase gene from Brevibacillus brevis. NIScPR-CSIR, New Delhi

Kumar S, Stecher G, Tamura K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33:1870–1874. https://doi.org/10.1093/molbev/msw054 PubMed DOI PMC

Kyeremeh AG, Kikumoto T, Chuang DY et al (2000) Biological control of soft rot of Chinese cabbage using single and mixed treatments of bacteriocin-producing avirulent mutants of Erwinia carotovora subsp. carotovora. J Gen Plant Pathol 66:264–268. https://doi.org/10.1007/PL00012957 DOI

Laskowski RA, MacArthur MW, Moss DS, Thornton JM (1993) PROCHECK: a program to check the stereochemical quality of protein structures. J Appl Crystallogr 26:283–291. https://doi.org/10.1107/s0021889892009944 DOI

Lin Y-H, Xu J-L, Hu J et al (2003) Acyl-homoserine lactone acylase from Ralstonia strain XJ12B represents a novel and potent class of quorum-quenching enzymes. Mol Microbiol 47:849–860. https://doi.org/10.1046/j.1365-2958.2003.03351.x PubMed DOI

Liu S, Tang Y (2016) Identification and characterization of a new enterobacter onion bulb decay caused by Lelliottia amnigena in China. Appl Microbiol Open Access. https://doi.org/10.4172/2471-9315.1000114 DOI

Lowry OH, RosebroughFarrRandall NJALRJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275. https://doi.org/10.1016/s0021-9258(19)52451-6 PubMed DOI

Lu S, Wang J, Chitsaz F et al (2020) CDD/SPARCLE: the conserved domain database in 2020. Nucleic Acids Res 48:D265–D268. https://doi.org/10.1093/nar/gkz991 PubMed DOI

Masyahit M, Sijam K, Awang Y, Satar MGM (2009) First report on bacterial soft rot disease on dragon fruit (Hylocereus spp.) caused by Enterobacter cloacae in Peninsular Malaysia. Int J Agric Biol 11:659–666

Moreira SI, da DutraRodrigues DCAC et al (2013) Fungi and bacteria associated with post-harvest rot of ginger rhizomes in Espírito Santo, Brazil. Trop Plant Pathol 38:218–226. https://doi.org/10.1590/S1982-56762013000300006 DOI

Nain Z, Adhikari UK, Abdulla F et al (2020) Computational prediction of active sites and ligands in different AHL quorum quenching lactonases and acylases. J Biosci 45:1–19. https://doi.org/10.1007/s12038-020-0005-1 DOI

Ngadze E, Coutinho TA, Van Der Waals JE (2010) Disease notes: first report of soft rot of potatoes caused by Dickeya dadantii in Zimbabwe. Plant Dis 94:1263. https://doi.org/10.1094/PDIS-05-10-0361 PubMed DOI

Osei R, Yang C, Cui L et al (2021) Salicylic acid effect on the mechanism of Lelliottia amnigena causing potato soft rot. Folia Hortic 33:376–389. https://doi.org/10.2478/fhort-2021-0029 DOI

Pan J, Huang T, Yao F et al (2008) Expression and characterization of aiiA gene from Bacillus subtilis BS-1. Microbiol Res 163:711–716. https://doi.org/10.1016/j.micres.2007.12.002 PubMed DOI

Park S-Y, Lee SJ, Oh T-K et al (2003) AhlD, an N-acylhomoserine lactonase in Arthrobacter sp., and predicted homologues in other bacteria. Microbiology 149:1541–1550. https://doi.org/10.1099/mic.0.26269-0 PubMed DOI

Pedroza CJ, Flórez AM, Ruiz OS, Orduz S (2014) Enzymatic hydrolysis of molecules associated with bacterial quorum sensing using an acyl homoserine lactonase from a novel Bacillus thuringiensis strain. Antonie Van Leeuwenhoek 105:253–264. https://doi.org/10.1007/s10482-013-0072-5 PubMed DOI

Pérombelon MCM (2002) Potato diseases caused by soft rot erwinias: an overview of pathogenesis. Plant Pathol 51:1–12 DOI

Popović T, Jelušić A, Milovanović P et al (2017) First report of Pectobacterium atrosepticum, causing bacterial soft rot on calla lily in Serbia. Plant Dis 101:2145. https://doi.org/10.1094/PDIS-05-17-0708-PDN DOI

Prazdnova EV, Gorovtsov AV, Vasilchenko NG et al (2022) Quorum-sensing inhibition by gram-positive bacteria. Microorganisms 10(2):350 PubMed DOI PMC

Pritchard L, Glover RH, Humphris S et al (2016) Genomics and taxonomy in diagnostics for food security: soft-rotting enterobacterial plant pathogens. Anal Methods 8:12–24. https://doi.org/10.1039/C5AY02550H DOI

Rashid M, Chowdhury M, Sultana N (2013) In-vitro screening of some chemicals and biocontrol agents against Erwinia carotovora subsp. carotovora, the causal agent of soft rot of potato (Solanum tuberosum). Agric 11:1–9. https://doi.org/10.3329/agric.v11i2.17480 DOI

Rehman ZU, Leiknes T (2018) Quorum-quenching bacteria isolated from red sea sediments reduce biofilm formation by Pseudomonas aeruginosa. Front Microbiol. https://doi.org/10.3389/fmicb.2018.01354 PubMed DOI PMC

Roy A, Kucukural A, Zhang Y (2010) I-TASSER: a unified platform for automated protein structure and function prediction. Nat Protoc 5:725–738. https://doi.org/10.1038/nprot.2010.5 PubMed DOI PMC

Rutherford ST, Bassler BL (2012) Bacterial quorum sensing: its role in virulence and possibilities for its control. Cold Spring Harb Perspect Med 2:a012427–a012427. https://doi.org/10.1101/cshperspect.a012427 PubMed DOI PMC

Sakr MM, AboshanabAboulwafaHassouna KMAMMNA-H (2013) Characterization and complete sequence of lactonase enzyme from Bacillus weihenstephanensis isolate P65 with potential activity against acyl homoserine lactone signal molecules. Biomed Res Int 2013:192589. https://doi.org/10.1155/2013/192589 PubMed DOI

Salem EA, Abd El-Shafea YM (2018) Biological control of potato soft rot caused by Erwinia carotovora subsp. carotovora. Egypt J Biol Pest Control 28:94. https://doi.org/10.1186/s41938-018-0100-x DOI

Sarfraz S, Sahi ST, Oulghazi S et al (2020) Species diversity of Dickeya and Pectobacterium causing potato blackleg disease in Pakistan. Plant Dis 104:1492–1499. https://doi.org/10.1094/PDIS-08-19-1743-RE PubMed DOI

Schroeder BK, Waters TD, Du Toit LJ (2010) Evaluation of onion cultivars for resistance to Enterobacter cloacae in storage. Plant Dis 94:236–243. https://doi.org/10.1094/PDIS-94-2-0236 PubMed DOI

Schuster M, Joseph Sexton D, Diggle SP, Peter Greenberg E (2013) Acyl-homoserine lactone quorum sensing: from evolution to application. Annu Rev Microbiol 67:43–63. https://doi.org/10.1146/annurev-micro-092412-155635 PubMed DOI

Sheladiya P, Kapadia C, Prajapati V et al (2022) Production, statistical optimization, and functional characterization of alkali stable pectate lyase of Paenibacillus lactis PKC5 for use in juice clarification. Sci Rep. https://doi.org/10.1038/s41598-022-11022-0 PubMed DOI PMC

Sohail M, Latif Z (2015) Phylogenetic analysis of polygalacturonase-producing Bacillus and Pseudomonas isolated from plant waste material. Jundishapur J Microbiol 9:1–6. https://doi.org/10.5812/jjm.28594 DOI

Suryanti E, Rusmana I, Wahyudi AT, Akhdiya A (2020) Characterization and expression of acyl homoserine lactone (AHL) lactonase in E. coli BL21 (DE3) pLysS. EurAsian J Biosci Eurasia J Biosci 14:6519–6526

Thomas PW, Stone EM, Costello AL et al (2005) The quorum-quenching lactonase from Bacillus thuringiensis is a metalloprotein. Biochemistry 44:7559–7569. https://doi.org/10.1021/bi050050m PubMed DOI

Toth IK, Bell KS, Holeva MC, Birch PRJ (2003) Soft rot erwiniae: from genes to genomes. Mol Plant Pathol 4:17–30. https://doi.org/10.1046/j.1364-3703.2003.00149.x PubMed DOI

Toth IK, van der Wolf JM, Saddler G et al (2011) Dickeya species: an emerging problem for potato production in Europe. Plant Pathol 60:385–399. https://doi.org/10.1111/j.1365-3059.2011.02427.x DOI

Tsai C-S, Winans SC (2010) LuxR-type quorum-sensing regulators that are detached from common scents. Mol Microbiol 77:1072–1082. https://doi.org/10.1111/j.1365-2958.2010.07279.x PubMed DOI PMC

Tsuda K, Tsuji G, Higashiyama M et al (2016) Biological control of bacterial soft rot in Chinese cabbage by Lactobacillus plantarum strain BY under field conditions. Biol Control 100:63–69. https://doi.org/10.1016/j.biocontrol.2016.05.010 DOI

Vinoj G, Vaseeharan B, Thomas S et al (2014) Quorum-quenching activity of the AHL-lactonase from Bacillus licheniformis DAHB1 inhibits vibrio biofilm formation in vitro and reduces shrimp intestinal colonisation and mortality. Mar Biotechnol (NY) 16:707–715. https://doi.org/10.1007/s10126-014-9585-9 DOI

Wang LH, Weng LX, Dong YH, Zhang LH (2004) Specificity and enzyme kinetics of the quorum-quenching N-acyl homoserine lactone lactonase (AHL-lactonase). J Biol Chem 279:13645–13651. https://doi.org/10.1074/jbc.M311194200 PubMed DOI

Wang N, Reidsma P, Pronk AA et al (2018) Can potato add to China’s food self-sufficiency? The scope for increasing potato production in China. Eur J Agron 101:20–29. https://doi.org/10.1016/j.eja.2018.07.002 DOI

Wang W-Z, Morohoshi T, Someya N, Ikeda T (2012) AidC, a novel N -acylhomoserine lactonase from the potato root-associated cytophaga-flavobacteria-bacteroides (CFB) group Bacterium Chryseobacterium sp. strain StRB126. Appl Environ Microbiol 78:7985–7992. https://doi.org/10.1128/AEM.02188-12 PubMed DOI PMC

Waters CM, Bassler BL (2005) Quorum sensing: cell-to-cell communication in bacteria. Annu Rev Cell Dev Biol 21:319–346. https://doi.org/10.1146/annurev.cellbio.21.012704.131001 PubMed DOI

Whiteley M, Diggle SP, Greenberg EP (2017) Progress in and promise of bacterial quorum sensing research. Nature 551:313–320. https://doi.org/10.1038/nature24624 PubMed DOI PMC

Wu J, Jiao Z, Guo F et al (2016) Constitutive and secretory expression of the AiiA in Pichia pastoris inhibits Amorphophallus konjac soft rot disease. Am J Mol Biol 06:79–87. https://doi.org/10.4236/ajmb.2016.62009 DOI

Yang J, Zhang Y (2015) I-TASSER server: new development for protein structure and function predictions. Nucleic Acids Res 43:174–181 DOI

Zhang C, Freddolino PL, Zhang Y (2017) COFACTOR: improved protein function prediction by combining structure, sequence and protein-protein interaction information. Nucleic Acids Res 45:W291–W299. https://doi.org/10.1093/nar/gkx366 PubMed DOI PMC

Zhang Y, Skolnick J (2004) Scoring function for automated assessment of protein structure template quality. Proteins 57:702–710. https://doi.org/10.1002/prot.20264 PubMed DOI