Abstract
Coral-associated microbes from Marine National Park (MNP), Gulf of Kutch (GoK), Gujarat, India, were screened for siderophore production. Maximum siderophore-producing isolate NP-C49 and its compound were identified and characterized. The isolate was identified as Klebsiella sp. through 16S rRNA genes sequencing (GenBank accession nos. KY412519 and MTCC 25160). Antibiotic susceptibility profile against 20 commercial antibiotics showed its more sensitivity compared to human pathogenic strain, i.e., Klebsiella pneumonia. The compound was identified as phenazine-1-carboxylic acid (PCA) using the multinuclear ID (1H and 13C) and 2D (1H–1H COSY and 1H–13C HETCOR) NMR along with high-resolution mass spectrometry. No significant difference in the bacterial growth in the presence of PCA, FeCl3 and Fe(OH)3 indicated involvement of factors other than PCA in bacterial growth. The study first reports the identification and characterization of PCA from Klebsiella sp. both from terrestrial and marine sources.
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References
Abraham A, Philip S, Jacob MK et al (2015) Phenazine-1-carboxylic acid mediated anti-oomycete activity of the endophytic Alcaligenes sp. EIL-2 against Phytophthora meadii. Microbiol Res 170:229–234. https://doi.org/10.1016/j.micres.2014.06.002
Ahmed E, Holmström SJM (2014) Siderophores in environmental research: roles and applications. Microb Biotechnol 7:196–208. https://doi.org/10.1111/1751-7915.12117
Bachman MA, Oyler JE, Burns SH et al (2011) Klebsiella pneumoniae yersiniabactin promotes respiratory tract infection through evasion of lipocalin 2. Infect Immun 79:3309–3316. https://doi.org/10.1128/iai.05114-11
Barman S, Hens DK, Koley H et al (2008) Chromosomal and plasmid encoded drug resistances of a Klebsiella pneumoniae UTI 2 strain isolated from urine of a post-operative patient. World J Microbiol Biotechnol 24:2693–2697. https://doi.org/10.1007/s11274-008-9798-z
Butler A (2005) Marine siderophores and microbial iron mobilization. Biometals 18:369–374. https://doi.org/10.1007/s10534-005-3711-0
Dutta S, Morang P, Nishanth Kumar S, Dileep Kumar BS (2014) Fusarial wilt control and growth promotion of pigeon pea through bioactive metabolites produced by two plant growth promoting rhizobacteria. World J Microbiol Biotechnol 30:1111–1121. https://doi.org/10.1007/s11274-013-1532-9
Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791. https://doi.org/10.1111/j.1558-5646.1985.tb00420.x
Gholia N, Toky V, Chhibber S (2004) Effect of antibacterials on surface properties and adhesion to uroepithelial cells in Klebsiella pneumoniae. World J Microbiol Biotechnol 20:775–779. https://doi.org/10.1007/s11274-004-5839-4
Govindarajan M, Kwon S-W, Weon H-Y (2007) Isolation, molecular characterization and growth-promoting activities of endophytic sugarcane diazotroph Klebsiella sp. GR9. World J Microbiol Biotechnol 23:997–1006. https://doi.org/10.1007/s11274-006-9326-y
Haldar S, Mody KH, Jha B (2011) Abundance, diversity and antibiotics resistance pattern of Vibrio spp. in coral ecosystem of Kurusadai island. J Basic Microbiol 51:153–162. https://doi.org/10.1002/jobm.201000153
Hoffmann LJ, Breitbarth E, Boyd PW, Hunter KA (2012) Influence of ocean warming and acidification on trace metal biogeochemistry. Mar Ecol Prog Ser 470:191–205. https://doi.org/10.3354/meps10082
Holden VI, Breen P, Houle S et al (2016) Klebsiella pneumoniae siderophores induce inflammation, bacterial dissemination, and HIF-1ɑ stabilization during pneumonia. mBio 7:e01397-16. https://doi.org/10.1128/mbio.01397-16
Höller C (2015) Incidence of Klebsiella species in surface waters and their expression of virulence factors incidence of Klebsiella species in surface waters and their expression of virulence factors. Am Soc Microbiol. https://doi.org/10.1128/AEM.67.7.3325
Jayatilake GS, Thornton MP, Leonard AC et al (1996) Metabolites from an antarctic sponge-associated bacterium, Pseudomonas aeruginosa. J Nat Prod 59:293–296. https://doi.org/10.1021/np960095b
Johnstone TC, Nolan EM (2015) Beyond iron: non-classical biological functions of bacterial siderophores. Dalton Trans 44:6320–6339. https://doi.org/10.1039/c4dt03559c
Kabir MM, Fakhruddin ANM, Chowdhury MAZ et al (2018) Isolation and characterization of chromium(VI)-reducing bacteria from tannery effluents and solid wastes. World J Microbiol Biotechnol 34:126. https://doi.org/10.1007/s11274-018-2510-z
Kennedy RK, Naik PR, Veena V et al (2015) 5-Methyl phenazine-1-carboxylic acid: a novel bioactive metabolite by a rhizosphere soil bacterium that exhibits potent antimicrobial and anticancer activities. Chem Biol Interact 231:71–82. https://doi.org/10.1016/j.cbi.2015.03.002
Li J, Zhang S, Shi S, Huo P (2011) Mutational approach for N2-fixing and P-solubilizing mutant strains of Klebsiella pneumoniae RSN19 by microwave mutagenesis. World J Microbiol Biotechnol 27:1481–1489. https://doi.org/10.1007/s11274-010-0600-7
Maatallah M, Vading M, Kabir MH et al (2014) Klebsiella variicola is a frequent cause of bloodstream infection in the Stockholm area, and associated with higher mortality compared to K. pneumoniae. PLoS ONE 9:e113539. https://doi.org/10.1371/journal.pone.0113539
Maddula VSRK, Pierson EA, Pierson LS (2008) Altering the ratio of phenazines in Pseudomonas chlororaphis (aureofaciens) strain 30–84: effects on biofilm formation and pathogen inhibition. J Bacteriol 190:2759–2766. https://doi.org/10.1128/JB.01587-07
Mansson M, Gram L, Larsen TO (2011) Production of bioactive secondary metabolites by marine Vibrionaceae. Mar Drugs 9:1440–1468. https://doi.org/10.3390/md9091440
Mudhulkar R, Nair RR, Raval IH et al (2017) Visualizing Zn2+ in living whole organism Artemia by a natural fluorimetric intermediate siderophore. ChemistrySelect 2:6407–6412. https://doi.org/10.1002/slct.201701071
Mudhulkar R, Rajapitamahuni S, Srivastava S et al (2018) Identification of a new siderophore acinetoamonabactin produced by a salt-tolerant bacterium acinetobacter soli. ChemistrySelect. https://doi.org/10.1002/slct.201801527
Nair RR, Raju M, Patel NP et al (2015) Naked eye instant reversible sensing of Cu2+ and its in situ imaging in live brine shrimp Artemia. Analyst 140:5464–5468
Nei M, Kumar S (2000) Molecular evolution and phylogenetics. Oxford University Press, Oxford
Patel NP, Haldar S (2019) Evaluation of traditional fish preservation method of Masmin from skipjack tuna (Katsuwonus pelamis) in Lakshadweep, India, with respect to nutritional and environmental perspectives. J Food Process Preserv. https://doi.org/10.1111/jfpp.14124
Patel NP, Kumar SB, Haldar S (2017) Role of bacteria in coral ecosystem. In: Kumar M, Ralph P (eds) Systems biology of marine ecosystems. Springer, Cham, pp 317–341
Penesyan A, Kjelleberg S, Egan S (2010) Development of novel drugs from marine surface associated microorganisms. Mar Drugs 8:438–459. https://doi.org/10.3390/md8030438
Pérez-Miranda S, Cabirol N, George-Téllez R, Zamudio-Rivera LS, Fernández FJ (2007) O-CAS, a fast and universal method for siderophore detection. J Microbiol Methods 70:127–131. https://doi.org/10.1016/j.mimet.2007.03.023
Pierson LS, Pierson EA (2010) Metabolism and function of phenazines in bacteria: impacts on the behavior of bacteria in the environment and biotechnological processes. Appl Microbiol Biotechnol 86:1659–1670. https://doi.org/10.1007/s00253-010-2509-3
Price-Whelan A, Dietrich LEP, Newman DK (2006) Rethinking “secondary” metabolism: physiological roles for phenazine antibiotics. Nat Chem Biol 2:71–78. https://doi.org/10.1038/nchembio764
Raina J, Tapiolas D, Motti C et al (2016) Isolation of an antimicrobial compound produced by bacteria associated with reef-building corals. PeerJ Prepr. https://doi.org/10.7717/peerj.2275
Raju M, Nair RR, Raval IH et al (2015) Reporting a new siderophore based Ca2+ selective chemosensor that works as a staining agent in the live organism Artemia. Analyst 140:7799–7809
Raju M, Patel TJ, Nair RR, Chatterjee PB (2016) Xanthurenic acid: a natural ionophore with high selectivity and sensitivity for potassium ions in an aqueous solution. New J Chem 40:1930–1934
Raju M, Srivastava S, Nair RR et al (2017) Siderophore coated magnetic iron nanoparticles: rational designing of water soluble nanobiosensor for visualizing Al3+ in live organism. Biosens Bioelectron 97:338–344. https://doi.org/10.1016/j.bios.2017.06.013
Rattanachuay P, Kantachote D, Tantirungkij M et al (2011) Antivibrio compounds produced by Pseudomonas sp. W3: characterisation and assessment of their safety to shrimps. World J Microbiol Biotechnol 27:869–880. https://doi.org/10.1007/s11274-010-0529-x
Rocha J, Peixe L, Gomes NCM, Calado R (2011) Cnidarians as a source of new marine bioactive compounds—an overview of the last decade and future steps for bioprospecting. Mar Drugs 9:1860–1886. https://doi.org/10.3390/md9101860
Schwyn B, Neilands JB (1987) Universal chemical assay for the detection and determination of siderophores. Anal Biochem 160:47–56. https://doi.org/10.1016/0003-2697(87)90612-9
Shanmugaiah V, Mathivanan N, Varghese B (2010) Purification, crystal structure and antimicrobial activity of phenazine-1-carboxamide produced by a growth-promoting biocontrol bacterium, Pseudomonas aeruginosa MML2212. J Appl Microbiol 108:703–711. https://doi.org/10.1111/j.1365-2672.2009.04466.x
Tamura K, Stecher G, Peterson D et al (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729. https://doi.org/10.1093/molbev/mst197
Wang Y, Newman DK (2008) Redox reactions of phenazine antibiotics with ferric (hydr)oxides and molecular oxygen. Environ Sci Technol 42:2380–2386. https://doi.org/10.1021/es702290a
Wang Y, Wilks JC, Danhorn T et al (2011) Phenazine-1-carboxylic acid promotes bacterial biofilm development via ferrous iron acquisition. J Bacteriol 193:3606–3617. https://doi.org/10.1128/JB.00396-11
Zhang L, Tian X, Kuang S et al (2017) Antagonistic activity and mode of action of phenazine-1-carboxylic acid, produced by marine bacterium Pseudomonas aeruginosa PA31x, against Vibrio anguillarum in vitro and in a zebrafish in vivo model. Front Microbiol. https://doi.org/10.3389/fmicb.2017.00289
Zhou L, Jiang H-X, Sun S et al (2016) Biotechnological potential of a rhizosphere Pseudomonas aeruginosa strain producing phenazine-1-carboxylic acid and phenazine-1-carboxamide. World J Microbiol Biotechnol 32:50. https://doi.org/10.1007/s11274-015-1987-y
Acknowledgements
NPP and MR has equally contributed to the manuscript. This manuscript has been assigned registration number CSIR‐CSMCRI—055/2019. We thank CSIR, Govt. of India for financial assistance as CSIR-SRF to MR (31/028(0218)/2017-EMR-I) and NPP (31/028(0202)/2015-EMR-I). We are also thankful to AESD&CIF of CSIR-CSMCRI for continued analytical and instrumentation facility.
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This article does not contain any studies with human participants or animals performed by any of the authors. Prior permission letter was obtained to collect water samples from coral surface from the MNP authorities with letter Reference No. B/WLP/T-5/472/473/2015-16.
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Patel, N.P., Raju, M., Haldar, S. et al. Characterization of phenazine-1-carboxylic acid by Klebsiella sp. NP-C49 from the coral environment in Gulf of Kutch, India. Arch Microbiol 202, 351–359 (2020). https://doi.org/10.1007/s00203-019-01742-9
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DOI: https://doi.org/10.1007/s00203-019-01742-9