Characterization and cytotoxicity assessment of biosurfactant derived from Lactobacillus pentosus NCIM 2912

戊糖乳杆菌 NCIM 2912 衍生的生物表面活性剂的表征及细胞毒性评估

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作者:Vikrant Sharma #, Deepti Singh #, Mehak Manzoor, Arun G Banpurkar, Surekha K Satpute, Deepansh Sharma

Abstract

Noteworthy properties of biosurfactant (BS) are fascinating scientific fraternity to explore them for food, medicinal, cosmetic, or pharmaceutical etc. applications. Newer products intended for pharmaceutical purposes are mandatory to go through pragmatic evaluation protocols. BS, being less cytotoxic, offers an ideal candidature for widespread applications in the healthcare sector. The goal of the current study was the isolation, physico-chemical characterization, and safety assessment of cell-associated biosurfactant (CABS) from Lactobacillus pentosus NCIM 2912. The culture was grown in a 3-L fermentor to produce CABS from the cell pellets through procedures like centrifugation, filtration, dialysis, column chromatography, and freeze-drying. Further, physical properties like surface tension (ST), critical micelle concentration (CMC), contact angle (CA), emulsification activity, stability of emulsion (height of emulsion, the extent of coalescence, and appearance), and ionic character of CABS were evaluated. Analytical characterization through TLC, FTIR, NMR, and GC-MS was carried out. The physico-chemical characterization revealed CABS as an anionic, multicomponent glycolipopeptide having a hydrophobic chain comprising butanoic acid (C4), decanoic acid (C10), undecanoic acid (C11), tridecanoic acid (C13), hexadecenoic acid (C16), and octadecanoic acid (C18). The oil-in-water (O/W) emulsions formed by CABS with various oils (olive, sesame, soybean, coconut) were stabilized up to the 7th day of storage and were analogous with polysorbate 80 (emulsifier/defoamer used in food industries). The O/W emulsions are quite stable at room temperature with no evidence of coalescence of droplets around 1 week. The cytotoxicity of CABS was evaluated through MTT (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide) assay. Cytotoxicity study performed on the human embryonic kidney (HEK 293), mouse fibroblast ATCC L929 and human epithelial type (HEP-2) cell lines recorded viability of 90.3 ± 0.1%, 99.2 ± 0.43, and 94.3 ± 0.2% respectively. The toxicity of the BS was comparable to that of the commercially used rhamnolipid sample. Thus, CABS derived from L. pentosus NCIM 2912 pose promising applications in the pharmaceutical, food industries acquiescently. The multifunctional potential of the incredibly versatile microbial product like BS from lactic acid bacteria (LAB) certainly contributes to wider avenues for varied industries.

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