Isolation and Molecular Characterization of Bacillus spp. Isolated from Soil for Production of Industrial Enzymes
Cumhur Avşar1*, Hilal Koyuncu1, E. Sümer Aras2
1. Department of Biology, Faculty of Science and Arts, Sinop University, 57000, Sinop, Turkey.
2. Department of Biology, Faculty of Science, Ankara University, 06100 Tandoğan, Ankara, Turkey.
Abstract: The main objective of our study was to investigate the morphologic and phenotypic properties, antibiotic resistance, random amplified polymorphic DNA (RAPD), enterobacterial repetitive intragenic consensus (ERIC) and Single-stranded conformation polymorphism (SSCP) profiles of the Bacillus spp. isolated from soil of Sinop environs. The aim of the investigation was also to check their ability for protease, amylase and cellulose enzymes production. A total of thirty-nine isolates were identified as Bacillus spp. based on morphological and physiological properties. The isolates were resistant on the percentage of 100% to penicillin; 66.6% to rifampicine; 23.7% to novobiocin; 48.7% to cefepime; 87.1% to ceftazidime; 89.7% to oxacillin; 2.5% to streptomycin; 30.7% to clindamycin; 7.6% to tetracycline; 12.8% to ampicillin/sulbactam; 10.2% to gentamicin; 56.4% to ceftriaxone; 84.6% to polymyxin B and 12.8% to amikacin were determined. In addition, all isolates were susceptible to imipenem, ciprofloxacin, meropenem and ofloxacin. All of the isolates produced at least two of the three enzymes, and 53.8% of strains were able to produce all three enzymes. The dendrograms generated by using M13-10, OPL3 RAPD primers, ERIC primers and P11/P13 SSCP primers revealed that the strains were separated to two major groups at similarity levels of 5%, 2%, 38% and 15% or above, respectively. As a result, SSCP, ERIC and RAPD analysis revealed 100% polymorphism indicating the strength of these methods in the differentiation of Bacillus isolates. In addition, it appears that Bacillus strains associated with soil are the source of industrial enzymes and may play an important role in enzyme catalyzed organic matter cycling in soil environments.
Keywords: Bacillus spp., SSCP, RAPD, ERIC-PCR, Bacterial enzymes.
Pages: 72 – 86 | Full PDF Paper