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 (Rf = 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.

Gene Expression and Characterization of Iturin A Lipopeptide Biosurfactant from Bacillus aryabhattai for Enhanced Oil Recovery

“…In previous studies, a lipopeptide showed 68 ± 0.5% emulsification activity (E24) [ 7 ]. A cell-free broth of B. velezensis showed 56.2% E24 with engine oil [ 31 ]. A biosurfactant from mutant A. niger M2 showed 62.30% of E24 with 59.81 cm 2 oil displacement area [ 32 ], and a 66.4 cm 2 oil displacement area was shown by a biosurfactant from R. arrhizus [ 33 ].…”

Tapping the Role of Microbial Biosurfactants in Pesticide Remediation: An Eco-Friendly Approach for Environmental Sustainability

“…Lipopeptides are the type of natural product produced by non-ribosomal peptide synthases (NRPS) ( De Giani et al, 2021 ), which are huge multifunctional enzyme clusters ( Williams and Trindade, 2017 ; Lamilla et al, 2021 ). Surfactin is considered one of the most potent biosurfactants ( Meena et al, 2021 ) composed of cyclic lipopeptides with seven amino acid ringed structures linked to a fatty acid chain through lactone linkage ( Figure 3 ; Onaizi, 2018 ; Sarwar et al, 2018 ). Table 1 summarizes the type of biosurfactant as well as the microbes that produce them and their roles in pesticide remediation.…”

Extraction, purification and applications of biosurfactants based on microbial-derived glycolipids and lipopeptides: a review

“…In size exclusion chromatography, several column volumes of sodium phosphate buffer (20 mM) were used to wash out the Sephadex G-25 packed matrix (pH 7.5). The DEAE active fraction with biosurfactant was put onto the surface of a Sephadex column(G-25) and eluted with 20 mM sodium phosphate buffer (pH 7.5), with fractions collected based on the molecular weight (Meena et al 2021). HPLC is developed to perform the qualitative and quantitative analysis of the biosurfactant like rhamnolipids and lipopeptides.…”

“…HPLC is developed to perform the qualitative and quantitative analysis of the biosurfactant like rhamnolipids and lipopeptides. Qualification and characterization of the biosurfactant can be executed by this method (Meena et al 2021).…”

“…After activation with 0.5 M NaOH, the diethyl aminoethyl (DEAE) cellulose-packed glass column was equilibrated with sodium phosphate buffer (20 mM, pH 7.5) and filled with five mL of biosurfactant(dialyzed) preparation (Meena et al 2020). With the help of a low ionic strength buffer, the unbound protein was washed out, and bound biosurfactant was eluted by stepwise gradient with different molarity of NaCl in sodium phosphate buffer (Meena et al 2021). Chromatographic way of purification gives highly purified product compared to other downstream processing methods, allowing the purification based on solubility, molecular weight, and ionic properties.…”