Dataset Information

Purification and identification of a surfactin biosurfactant and engine oil degradation by Bacillus velezensis KLP2016.

ABSTRACT: Engine oil used in automobiles is a threat to soil and water due to the recalcitrant properties of its hydrocarbons. It pollutes surrounding environment which affects both flora and fauna. Microbes can degrade hydrocarbons containing engine oil and utilize it as a substrate for their growth. Our results demonstrated that cell-free broth of Bacillus velezensis KLP2016 (Gram?+?ve, endospore forming; Accession number KY214239) recorded an emulsification index (E₂₄%) from 52.3% to 65.7% against different organic solvents, such as benzene, pentane, cyclohexane, xylene, n-hexane, toluene and engine oil. The surface tension of the cell-free broth of B. velezensis grown in Luria-Bertani broth at 35 °C decreased from 55 to 40 mN m^-1at critical micelle concentration 17.2 µg/mL. The active biosurfactant molecule of cell-free broth of Bacillus velezensis KLP2016 was purified by Dietheylaminoethyl-cellulose and size exclusion chromatography, followed by HPLC (RT?=?1.130), UV-vis spectrophotometry (210 nm) and thin layer chromatography (R_f?=?0.90). The molecular weight of purified biosurfactant was found to be?~?1.0 kDa, based on Electron Spray Ionization-MS. A concentration of 1980?×?10^-2 parts per million of CO₂ was trapped in a KOH solution after 15 days of incubation in Luria-Bertani broth containing 1% engine oil. Our results suggest that bacterium Bacillus velezensis KLP2016 may promise a new dimension to solving the engine oil pollution problem in near future.

SUBMITTER: Meena KR

PROVIDER: S-EPMC7844888 | biostudies-literature | 2021 Jan

REPOSITORIES: biostudies-literature

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Purification and identification of a surfactin biosurfactant and engine oil degradation by Bacillus velezensis KLP2016.

Meena Khem Raj KR Dhiman Rajni R Singh Kailash K Kumar Sachin S Sharma Abhishek A Kanwar Shamsher S SS Mondal Rittick R Das Sandip S Franco Octavio L OL Mandal Amit Kumar AK

Microbial cell factories 20210128 1

Engine oil used in automobiles is a threat to soil and water due to the recalcitrant properties of its hydrocarbons. It pollutes surrounding environment which affects both flora and fauna. Microbes can degrade hydrocarbons containing engine oil and utilize it as a substrate for their growth. Our results demonstrated that cell-free broth of Bacillus velezensis KLP2016 (Gram + ve, endospore forming; Accession number KY214239) recorded an emulsification index (E<sub>24</sub>%) from 52.3% to 65.7% a ...[more]

PMID: 33509221

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Project description:Surfactants can reduce the interfacial surface tension between two immiscible liquids making them a desirable component for various industrial applications. However, the toxic nature of chemical surfactants brought immense attention towards biosurfactants. Being biodegradable, biosurfactants are eco-friendly and considered safer for different commercial uses. This study focused on the production of biosurfactant from an oil-degrading bacteria and its functional efficacy for prospective industrial applications. Here, a promising oil-tolerant strain, Bacillus velezensis S2 was isolated from oil contaminated sites which showed >50% degradation of convoluted crude oil within 28 days in comparison to a control. The isolate was then found to produce an excellent surface-active compound with an emulsification index of 67.30 ± 0.8% and could reduce the surface tension up to 36.86 ± 0.36 mN m-1. It also manifested a critical micelle concentration of 45 mg L-1 while reducing the surface tension from 72 to 30 mN m-1. When extracting biosurfactant from isolated bacteria, ethyl acetate extraction showed 1.5 times greater efficacy than chloroform : methanol extraction. The purified biosurfactant was characterized using TLC, 1H NMR, 13C NMR, FTIR, elemental analyses and spectrophotometric techniques leading to its identification as a rhamnolipid. The stability of produced biosurfactant at higher temperature (up to 180 °C) was determined by thermal analysis, endorsing its application in high temperature reservoir conditions. Additionally, the extracted biosurfactant showed excellent foaming efficacy with insignificant antibacterial and cytotoxic responses, which indicates their potential application in cleaning and cosmetics industries. Thus, the present study outlines a bi-functional novel isolate Bacillus velezensis S2 which could play a significant role in oil remediation from the environment as well as serve as a potential source of non-toxic and eco-friendly biosurfactants for various industrial applications.

Project description:BackgroundBiosurfactants, being highly biodegradable, ecofriendly and multifunctional compounds have wide applications in various industrial sectors including environmental bioremediation. Surfactin, a member of lipopeptide family, which is considered as one of the most powerful biosurfactants due to its excellent emulsifying activities as well as environmental and therapeutic applications. Therefore, the aim of this study was to investigate the newly isolated bacterial strain S2MT for production of surfactin-like biosurfactants and their potential applications for oil-contaminated soil remediation.ResultsIn this study, the strain S2MT was isolated from lake sediment and was identified as Bacillus nealsonii based on transmitted electron microscopy (TEM) and 16S rRNA ribo-typing. The strain S2MT produced biosurfactant that reduced the surface tension (34.15 ± 0.6 mN/m) and displayed excellent emulsifying potential for kerosene (55 ± 0.3%). Additionally, the maximum biosurfactant product yield of 1300 mg/L was achieved when the composition of the culture medium was optimized through response surface methodology (RSM). Results showed that 2% glycerol and 0.1% NH4NO3 were the best carbon/nitrogen substrates for biosurfactant production. The parameters such as temperature (30 °C), pH (8), agitation (100 rpm), NH4NO3 (0.1%) and NaCl (0.5%) displayed most significant contribution towards surface tension reduction that resulted in enhanced biosurfactant yield. Moreover, the extracted biosurfactants were found to be highly stable at environmental factors such as salinity, pH and temperature variations. The biosurfactants were characterized as cyclic lipopeptides relating to surfactin-like isoforms (C13-C15) using thin-layer chromatography (TLC), Ultra high performance liquid chromatography and mass spectrometry (UHPLC-MS). The crude biosurfactant product displayed up to 43.6 ± 0.08% and 46.7 ± 0.01% remediation of heavy engine-oil contaminated soil at 10 and 40 mg/L concentrations, respectively.ConclusionPresent study expands the paradigm of surfactin-like biosurfactants produced by novel isolate Bacillus nealsonii S2MT for achieving efficient and environmentally acceptable soil remediation as compared to synthetic surfactants.

Project description:BackgroundBacillus velezensis is an endospore-forming, free-living soil bacterium with potential as a biopesticide against a broad spectrum of microbial pathogens of plants. Its potential for commercial development is enhanced by rapid replication and resistance to adverse environmental conditions, typical of Bacillus species. However, the use of beneficial microbes against phytopathogens has not gained dominance due to limitations that may be overcome with new biopesticidal strains and/or new biological knowledge.ResultsHere, we isolated B. velezensis strain 9D-6 and showed that it inhibits the in vitro growth of prokaryotic and eukaryotic pathogens, including the bacteria Bacillus cereus , Clavibacter michiganensis, Pantoea agglomerans, Ralstonia solanacearum, Xanthomonas campestris, and Xanthomonas euvesicatoria; and the fungi Alternaria solani, Cochliobolus carbonum, Fusarium oxysporum, Fusarium solani, Gibberella pulicaris, Gibberella zeae, Monilinia fructicola, Pyrenochaeta terrestris and Rhizoctonia solani. Antimicrobial compounds with activity against Clavibacter michiganensis were isolated from B. velezensis 9D-6 and characterized by high resolution LC-MS/MS, yielding formulae of C52H91N7O13 and C53H93N7O13, which correspond to [Leu7] surfactins C14 and C15 (also called surfactin B and surfactin C), respectively. We further sequenced the B. velezensis 9D-6 genome which consists of a single circular chromosome and revealed 13 gene clusters expected to participate in antimicrobial metabolite production, including surfactin and two metabolites that have not typically been found in this species - ladderane and lantipeptide. Despite being unable to inhibit the growth of Pseudomonas syringae DC3000 in an in vitro plate assay, B. velezensis 9D-6 significantly reduced root colonization by DC3000, suggesting that 9D-6 uses methods other than antimicrobials to control phytopathogens in the environment. Finally, using in silico DNA-DNA hybridization (isDDH), we confirm previous findings that many strains currently classified as B. amyloliquefaciens are actually B. velezensis.ConclusionsThe data presented here suggest B. velezensis 9D-6 as a candidate plant growth promoting bacterium (PGPB) and biopesticide, which uses a unique complement of antimicrobials, as well as other mechanisms, to protect plants against phytopathogens. Our results may contribute to future utilization of this strain, and will contribute to a knowledge base that will help to advance the field of microbial biocontrol.

Dataset Information

Purification and identification of a surfactin biosurfactant and engine oil degradation by Bacillus velezensis KLP2016.

Publications

Purification and identification of a surfactin biosurfactant and engine oil degradation by Bacillus velezensis KLP2016.

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