可能产生表面活性剂的芽孢杆菌Sp MN 243657的多样性、分布、GC-MS分析及抗菌研究

C. Brinda, R. R. R. Ragunathan, Jesteena Johney
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引用次数: 0

摘要

生物表面活性剂是微生物产生的表面活性化合物。它们是两亲分子,具有亲水性和疏水性区域。由于生物表面活性剂无毒、可生物降解,对它们的需求呈指数级增长。它们在几个工业部门有不同的应用。本研究的目的是从印度喀拉拉邦和泰米尔纳德邦不同地区的石油污染土壤中分离和鉴定产生生物表面活性剂的原生细菌。土壤样本是从汽油泵和车间收集的,那里的土壤受到汽油、柴油和机油的污染。从污染土壤样品中分离得到该细菌,确定其为芽孢杆菌。采用不同的筛选技术,如血液溶血试验、油扩散试验、乳化能力试验、细菌对碳氢化合物的粘附活性等,对培养物进行了生物表面活性剂的筛选。利用分子技术筛选和鉴定了具有生产生物表面活性剂潜力的培养菌,并将其提交至NCBI基因库(MN 243657 - Bacillus sp)。将所选细菌用于生物表面活性剂的生产,并通过紫外、薄层色谱、红外光谱和质谱分析对其进行了表征。经薄层色谱测定,所得生物表面活性剂的Rf值为0.68。在紫外可见光谱研究中,分离的生物表面活性剂在415 nm处显示出峰值。FTIR分析表明,分离得到的生物表面活性剂在3340 cm -1处有一个强烈的峰。气相色谱-质谱分析表明,在不同的保留时间(12.75、10.22、4.98和3.87)下,生物表面活性剂的峰均为大峰。该生物表面活性剂对铜绿假单胞菌、大肠杆菌、肺炎克雷伯菌、金黄色葡萄球菌及真菌黑曲霉、地曲霉、黄曲霉等病原菌具有抑菌抑菌活性。
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Diversity and Distribution of Potential Biosurfactant Producing Bacillus Sp MN 243657, GC-MS Analysis and its Antimicrobial Study
Biosurfactants are microbially produced surface-active compounds. They are amphiphilic molecules with hydrophilic and hydrophobic regions. The demand for biosurfactants has been exponentially growing as they are nontoxic and biodegradable. They have different applications in several industrial sectors. The objective of this study was to isolate and characterize the native bacteria which produce biosurfactants from oil contaminated soil of different places in Kerala and Tamil Nadu, India. The soil samples were collected from petrol pumps and workshops where the soil is contaminated with petrol, diesel and oil. The bacteria were isolated from contaminated soil samples and confirmed as Bacillus sp.The cultures were screened for biosurfactant production by different screening techniques such as blood hemolysis, oil spreading assay, emulsification ability assay, bacterial adherence to hydrocarbons activity. The potential biosurfactant producing culture was selected and identified using molecular techniques and submitted to NCBI Gene Bank (MN 243657 – Bacillus sp). The selected bacterial culture was used for biosurfactant production and these were characterized by UV, TLC, FTIR and GC -MS analysis. The derived biosurfactant's Rf value was 0.68 as determined by a TLC chromatogram. In a UV-visible spectroscopy study, the isolated biosurfactant displayed a highest peak at 415 nm. According to FTIR analysis, the isolated biosurfactant displayed an intense peak at 3340 cm -1. The large peaks of the biosurfactant were observed at various retention times of 12.75, 10.22, 4.98, and 3.87, respectively, after GC-MS analysis. Antibacterial and antifungal activity of the biosurfactant was identified against pathogenic bacteria such as P.aeruginosa, E. coli, K. pneumoniae, S. aureus and fungi Aspergillus niger, Aspergillus terreus, Aspergillus flavus.
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