马库尔迪市污染土壤中产生物表面活性剂芽孢杆菌的筛选与鉴定

W. John, I. O. Ogbonna, G. Gberikon, C. Iheukwumere
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引用次数: 1

摘要

微生物合成的生物表面活性剂具有多种化学性质,由于其表面和界面张力降低活性而在工业上引起了人们的兴趣。本研究对污染土壤中的芽孢杆菌进行了筛选和鉴定,用于生产生物表面活性剂。这项研究是在尼日利亚马库尔迪联邦农业大学微生物实验室进行的。从马库尔迪市煤油店、棕榈油店、附近餐馆、机械车间和屠宰场污水污染的土壤样本中分离出芽孢杆菌。采用不同的筛选方法(油扩散法、β溶血法、滴塌法和乳化指数法)对芽孢杆菌进行了生物表面活性剂生产潜力的筛选。利用特异性引物在芽孢杆菌分离株中扩增srfAA (surfactin gene)基因,并在南非Inqaba Biotec公司测定其核苷酸序列。筛选结果采用方差分析(ANOVA)进行统计分析,置信度为95%。分离物RT7(4)B具有生产生物表面活性剂的能力,其乳化指数最高(E24)为73.25%,分离物PO7(3)C的排油率最高,为6.77 mm。从分离物RT7(4)B中获得的上清显示表面张力降低高达30.26 mN/m。该菌株对生物表面活性剂的生产有积极的影响,并对其进行了滴缩和溶血试验。聚合酶链反应(PCR)结果显示,从7株分离物中扩增出srfAA基因。基于这些发现,本研究中使用的分离物可以用于生物表面活性剂的生产,也可以用于生物修复和工业生物技术应用。关键词:生物表面活性剂;乳化指数;芽孢杆菌;表面张力;下降崩溃
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Screening and Characterization of Biosurfactant-Producing Bacillus Species Isolated from Contaminated Soils in Makurdi Metropolis
Biosurfactants synthesized by microorganisms are chemically diverse and have gained interest industrially due to their surface and interfacial tensions-reducing activities. In this study Bacillus species from contaminated soils were screened and characterized for biosurfactant production. The study was carried out at the Microbiology Laboratory, Federal University of Agriculture Makurdi, Nigeria. The Bacillus species were isolated from kerosene shops, palm oil shops, nearby restaurants, mechanic workshops and abattoir effluents- contaminated soil samples collected from Makurdi metropolis. The Bacillus spp. were screened for biosurfactants production potentials using various screening methods (oil spreading, beta haemolysis, drop collapse and emulsification index). Specific primers were used to amplify the srfAA (surfactin gene) gene in the Bacillus isolates and the nucleotide sequences were determined at Inqaba Biotec, South Africa. The screening results were statistically analysed using analysis of variance (ANOVA) at 95 % confidence level. Isolate RT7(4)B exhibited the ability to produce biosurfactant, as well as the highest emulsification index (E24) of 73.25 % while isolate PO7(3)C gave the highest oil displacement of 6.77 mm. The supernatant obtained from isolate RT7(4)B showed reduction in surface tension of up to 30.26 mN/m. The isolates gave positive results for biosurfactant production when subjected to drop collapse and Beta haemolytic tests. The Polymerase chain reaction (PCR) results revealed amplifications of srfAA gene from 7 isolates. Based on these findings, the isolates used in this study can be utilized for biosurfactant production, and can also be useful for bioremediation and industrial biotechnology applications. Keywords: Biosurfactants; emulsification index; Bacillus; surface tension; Drop collapse
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