Antifungal Activity of Rhamnolipid Biosurfactant Produced by Pseudomonas aeruginosa A4 against Plant Pathogenic Fungi

Authors

  • Theerawat Onlamool Center of Excellence in Innovative Biotechnology for Sustainable Utilization of Bioresources, Faculty of Agro-Industry, Prince of Songkla University, Songkhla 90110, Thailand
  • Atipan Saimmai Faculty of Agricultural Technology, Phuket Rajabhat University, Phuket 83000, Thailand
  • Suppasil Maneerat Center of Excellence in Innovative Biotechnology for Sustainable Utilization of Bioresources, Faculty of Agro-Industry, Prince of Songkla University, Songkhla 90110, Thailand

DOI:

https://doi.org/10.48048/tis.2023.6524

Keywords:

Rhamnolipid, Biosurfactant, Soapstock, Antifungal activity, Oil palm seed

Abstract

Biosurfactants have been shown to have a variety of other agricultural applications. Antimicrobial activity is a desirable property in a variety of biosurfactants. Several biosurfactants produced from bacteria have antibacterial activity against plant diseases, making them a promising biocontrol molecule for ensuring agricultural sustainability in the long run. The purpose of this research was to characterize biosurfactant produced at different carbon source by biosurfactant-producing bacterial strains isolated from contaminated soapstock sediment, as well as its antifungal properties. Biosurfactant-producing bacteria were isolated from soapstock-contaminated soil which bacteria were selected by using drop collapse and oil displacement tests. Twelve isolates reduced surface tension of culture broth from 50 to 31 - 43 mN/m. According to 16S rRNA sequence analysis, these isolates belong to 8 different genera (Acinetobacter, Citrobacter, Enterobacter, Klebsiella, Pantoea, Pseudomonas, Stenotrophomonas and Xanthomonas). The supernatant of Pseudomonas aeruginosa A4 grown in MSM supplemented with soapstock was the most effective biosurfactant against Aspergillus flavus F2, Aspergillus niger F14, Cunninghamella bertholletiae F1 and Rhizopus oryzae F5, inhibiting mycelium growth by 54, 61 59 and 50 %, respectively. The extract substance inhibited spore germination against A. flavus F2 and R. oryzae F5 with a minimum inhibitory concentration of 2.75 mg/mL. TLC, FT-IR, ESI-MS and GC-MS analysis demonstrated that the mono-rhamnolipids and di-rhamnolipids had the same 3-hydroxy fatty acid composition of C8, C10 and C12. Overall, the biosurfactant-producing strains identified in this investigation show promise for future development and application in cost-effective industrial-scale biotechnological processes.

HIGHLIGHTS

  • Biosurfactant producing bacteria were isolated from soapstock contaminated soil
  • The supernatant of Pseudomonas aeruginosa A4 grown in MSM supplemented with soapstock was the most effective biosurfactant against plant pathogenic fungi
  • Biosurfactant from Pseudomonas aeruginosa A4 was identified as rhamnolipid comprising of both mono and di-rhamnolipid congeners


GRAPHICAL ABSTRACT 

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References

M Hassanshahian. Isolation and characterization of biosurfactant producing bacteria from Persian Gulf (Bushehr provenance). Mar. Pollut. Bull. 2014; 86, 361-6.

P Das, S Mukherjee and R Sen. Improved bioavailability and biodegradation of a model polyaromatic hydrocarbon by a biosurfactant producing bacterium of marine origin. Chemosphere 2008; 72, 1229-34.

MJ Haas. Improving the economics of biodiesel production through the use of low value lipids as feedstocks: vegetable oil soapstock. Fuel. Process. Technol. 2005; 86, 1087-96.

R Piloto-Rodríguez, EA Melo, L Goyos-Pérez and S Verhelst. Conversion of by-products from the vegetable oil industry into biodiesel and its use in internal combustion engines: a review. Braz. J. Chem. Eng. 2014; 31, 287-301.

D Kitamoto, H Isoda and T Nakahara. Functions and potential applications of glycolipid biosurfactants-from energy-saving materials to gene delivery carriers. J. Biosci. Bioeng. 2002; 94, 187-201.

B Yin, J-D Gu and N Wan. Degradation of indole by enrichment culture and Pseudomonas aeruginosa Gs isolated from mangrove sediment. Int. Biodeterior. Biodegrad. 2005; 56, 243-8.

JD Thompson, DG Higgins and TJ Gibson. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic. Acids. Res. 1994; 22, 4673-80.

S-C Lee, S-J Lee, S-H Kim, I-H Park, Y-S Lee, S-Y Chung and Y-L Choi. Characterization of new biosurfactant produced by Klebsiella sp. Y6-1 isolated from waste soybean oil. Bioresour. Technol. 2008; 99, 2288-92.

A Saimmai, T Onlamool, V Sobhon and S Maneerat. An efficient biosurfactant-producing bacterium Selenomonas ruminantium CT2, isolated from mangrove sediment in south of Thailand. World J. Microbiol. Biotechnol. 2013; 29, 87-102.

D Goswami, SN Borah, J Lahkar, PJ Handique and S Deka. Antifungal properties of rhamnolipid produced by Pseudomonas aeruginosa DS9 against Colletotrichum. falcatum. J. Basic. Microbiol. 2015; 55, 1265-74.

G Ebadzadsahrai, EA Higgins Keppler, SD Soby and HD Bean. Inhibition of Fungal Growth and Induction of a Novel Volatilome in Response to Chromobacterium vaccinii Volatile Organic Compounds. Front. Microbiol. 2020; 11, 1035.

RA Slawecki, EP Ryan and DH Young. Novel fungitoxicity assays for inhibition of germination-associated adhesion of Botrytis cinerea and Puccinia recondita spores. Appl. Environ. Microbiol. 2002; 68, 597-601.

R Bhat, S Hathwar, R Hegde and A Kush. Exploration on production of rhamnolipid biosurfactants using native Pseudomonas aeruginosa strains. J. Biosci. Biotechnol. 2015; 4, 157-66.

MA Churchward, DM Brandman, T Rogasevskaia and JR Coorssen. Copper (II) sulfate charring for high sensitivity on-plate fluorescent detection of lipids and sterols: quantitative analyses of the composition of functional secretory vesicles. J. Chem. Biol. 2008; 1, 79-87.

B Thanomsub, T Watcharachaipong, K Chotelersak, P Arunrattiyakorn, T Nitoda and H Kanzaki. Monoacylglycerols: glycolipid biosurfactants produced by a thermotolerant yeast, Candida ishiwadae. J. Appl. Microbiol. 2004; 96, 588-92.

K Ichihara and Y Fukubayashi. Preparation of fatty acid methyl esters for gas-liquid chromatography. J. Lipid. Res. 2010; 51, 635-40.

AA Bodour and RM Miller-Maier. Application of a modified drop-collapse technique for surfactant quantitation and screening of biosurfactant-producing microorganisms. J. Microbiol. Methods. 1998; 32, 273-80.

NH Youssef, KE Duncan, DP Nagle, KN Savage, RM Knapp and MJ McInerney. Comparison of methods to detect biosurfactant production by diverse microorganisms. J. Microbiol. Meth. 2004; 56, 339-47.

YH Wei, CL Chou and JS Chang. Rhamnolipid production by indigenous Pseudomonas aeruginosa J4 originating from petrochemical wastewater. Biochem. Eng. J. 2005; 27, 146-54.

GS Kiran, TA Thomas and J Selvin. Production of a new glycolipid biosurfactant from marine Nocardiopsis lucentensis MSA04 in solid-state cultivation. Colloids. Surf. B. Biointerfaces 2010; 78, 8-16.

PG Carrillo, C Mardaraz, SI Pitta-Alvarez and AM Giulietti. Isolation and selection of biosurfactant-producing bacteria. World J. Microbiol. Biotechnol. 1996; 12, 82-4.

SB Batista, AH Mounteer, FR Amorim and MR Tótola. Isolation and characterization of biosurfactant/bioemulsifier-producing bacteria from petroleum contaminated sites. Bioresour Technol. 2006; 97, 868-75.

A Dadrasnia and S Ismail. Biosurfactant production by Bacillus salmalaya for lubricating oil solubilization and biodegradation. Int. J. Environ. Res. Public. Health. 2015; 12, 9848-63.

M Morikawa, Y Hirata and T Imanaka. A study on the structure - function relationship of lipopeptide biosurfactants. Biochim. Biophys. Acta. BBA - Mol. Cell. Biol. Lipids 2000; 1488, 211-8.

DG Cooper. Biosurfactants. Microbiol. Sci. 1986; 3, 145-9.

M Abouseoud, R Maachi, A Amrane, S Boudergua and A Nabi. Evaluation of different carbon and nitrogen sources in production of biosurfactant by Pseudomonas fluorescens. Desalination 2008; 223, 143-51.

CN Mulligan and BF Gibbs. Correlation of nitrogen metabolism with biosurfactant production by Pseudomonas aeruginosa. Appl. Environ. Microbiol. 1989; 55, 3016-9.

A Saimmai, O Rukadee, T Onlamool, V Sobhon and S Maneerat. Isolation and functional characterization of a biosurfactant produced by a new and promising strain of Oleomonas sagaranensis AT18. World J. Microbiol. Biotechnol. 2012; 28, 2973-86.

PA Willumsen and U Karlson. Screening of bacteria, isolated from PAH-contaminated soils, for production of biosurfactants and bioemulsifiers. Biodegradation 1996; 7, 415-23.

IV Nwaguma, CB Chikere and GC Okpokwasili. Isolation, characterization, and application of biosurfactant by Klebsiella pneumoniae strain IVN51 isolated from hydrocarbon-polluted soil in Ogoniland, Nigeria. Bioresour. Bioprocess. 2016; 3, 40.

F Yan, S Xu, J Guo, Q Chen, Q Meng and X Zheng. Biocontrol of post-harvest Alternaria alternata decay of cherry tomatoes with rhamnolipids and possible mechanisms of action. J. Sci. Food. Agric. 2015; 95, 1469-74.

AI Rodrigues, EJ Gudiña, L Abrunhosa, AR Malheiro, R Fernandes, JA Teixeira and LR Rodrigues. Rhamnolipids inhibit aflatoxins production in Aspergillus flavus by causing structural damages in the fungal hyphae and down-regulating the expression of their biosynthetic genes. Int. J. Food. Microbiol. 2021; 348, 109207.

P Thakur, NK Saini, VK Thakur, VK Gupta, RV Saini and AK Saini. Rhamnolipid the glycolipid biosurfactant: emerging trends and promising strategies in the field of biotechnology and biomedicine. Microb. Cell Factories 2021; 20, 1.

TAA Moussa, MS Mohamed and N Samak. Production and characterization of di-rhamnolipid produced by Pseudomonas aeruginosa TMN. Braz. J. Chem. Eng. 2014; 31, 867-80.

J Kiefer, MN Radzuan and J Winterburn. Infrared spectroscopy for studying structure and aging effects in rhamnolipid biosurfactants. Appl. Sci. 2017; 7, 533.

JF Liu, G Wu, SZ Yang and BZ Mu. Structural characterization of rhamnolipid produced by Pseudomonas aeruginosa strain FIN2 isolated from oil reservoir water. World J. Microbiol. Biotechnol. 2014; 30, 1473-84.

AM Abdel-Mawgoud, F Lépine and E Déziel. Rhamnolipids: diversity of structures, microbial origins and roles. Appl. Microbiol. Biotechnol. 2010; 86, 1323-36.

N Lourith and M Kanlayavattanakul. Natural surfactants used in cosmetics: Glycolipids. Int. J. Cosmet. Sci. 2009; 31, 255-61.

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Published

2022-12-28