Amity Institute of Microbial Technology, Amity University Rajasthan, Jaipur, Rajasthan, 303002, India.
Department of Physics, Savitribai Phule Pune University, Pune, 411007, India.
Braz J Microbiol. 2022 Mar;53(1):327-340. doi: 10.1007/s42770-021-00654-5. Epub 2021 Nov 24.
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.
生物表面活性剂（BS）的显著特性引起了科学界的极大兴趣，使其成为食品、医药、化妆品和制药等领域的研究对象。新型药物产品必须经过实用评估方案。由于 BS 的细胞毒性较低，因此成为医疗保健领域广泛应用的理想候选物。本研究的目的是从植物乳杆菌 NCIM 2912 中分离、理化特性分析和安全性评估细胞相关生物表面活性剂（CABS）。该培养物在 3-L 发酵罐中生长，通过离心、过滤、透析、柱层析和冷冻干燥等步骤从细胞沉淀中产生 CABS。进一步评估了表面张力（ST）、临界胶束浓度（CMC）、接触角（CA）、乳化活性、乳液稳定性（乳液高度、聚结程度和外观）和 CABS 的离子特性等物理性质。通过 TLC、FTIR、NMR 和 GC-MS 进行分析表征。理化特性分析表明，CABS 是一种阴离子、多组分糖脂肽，其疏水链由丁酸（C4）、癸酸（C10）、十一酸（C11）、十三酸（C13）、十六烯酸（C16）和十八酸（C18）组成。CABS 与各种油（橄榄油、芝麻油、大豆油、椰子油）形成的油包水（O/W）乳液在储存的第 7 天仍保持稳定，与聚山梨酯 80（食品工业中使用的乳化剂/消泡剂）类似。在室温下，O/W 乳液相当稳定，大约 1 周内没有观察到液滴聚结的证据。通过 MTT（3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四唑溴盐）测定法评估 CABS 的细胞毒性。在人胚肾（HEK 293）、小鼠成纤维细胞 ATCC L929 和人上皮型（HEP-2）细胞系上进行的细胞毒性研究记录了 90.3±0.1%、99.2±0.43%和 94.3±0.2%的活力。BS 的毒性与商业使用的鼠李糖脂样品相当。因此，从植物乳杆菌 NCIM 2912 中提取的 CABS 在制药和食品工业中具有广阔的应用前景。乳酸细菌（LAB）产生的多功能生物表面活性剂等微生物产品的多功能性确实为各个行业开辟了更广泛的途径。
