Studying of some metabolic activities, plasmid DNA and protein profiles of Bacillus bacteria isolated from the soil

In the study, a total of thirty two strains belonging to Bacillus brevis, Bacillus sphaericus, Bacillus cereus, Bacillus megaterium, Bacillus circulans, Bacillus subtilis, Bacillus licheniformis and Bacillus coagulans were analysed. Twenty nine of them were isolated from the soil. These strains isolated were physiologically and biochemically tested and described. Total cell protein profiles of bacteries were analysed by sodium dodecyl sulphate poh/acrylamide gel electrophoresis (SDS-PAGE). The moleculer weights of proteins were found ranging between 19 and 160 kD. SDS-PAGE analysis showed that those strains belong to the same category had similar protein profiles and that they were different from the others. Poh/~P-hydroxybutyrate (PHB) productions of Bacillus bacteria were determined by spectrophotometry method and it was found that PHB productions ranged from 1,06-41,67% depending on the of dry cell weight. Bacillus brevis M6 strain which has the highest PHB production (41,67%) was analysed in terms of its PHB production under certain conditions such as carbon, phosphate, nitrogen resources and oxygen limitations. It was found that the PHB production increased in the medium with citrat as carbon resource (51,85%) and in the medium with sodium nitrate as nitrogen resource (60,00%).However maximum PHB production was in the medium with phosphate (6,50%). PHB production in the medium without nitrogen and limited oxygen medium were 25,45% and 7,53% respectively. Economical production of PHB was investigated by using sugar beet molasses an the effects of different concentration molasses on PHB production were investigated. For this reason, Bacillus cereus M5 strain were selected and cultivated in l%-5% molasses concentration. It was found that strain was produced higher PHB (73,84%) in 1% molasses concentration. It was also observed that PHB productivity decreased with to the increaseing molasses concentration. The general inhibition of Bacillus isolates on the pathogen and contaminant test bacteria were investigated by using the agar diffusion method. As a result of this study, the following data were found: B. brevis M6 showed inhibition effect on Staphylococus aureus ATCC 25923 and on Bacillus thuringiensis RSKK 380; B. cereus M15 on Pseudomonas fluorescens RSKK 240, on Micrococcus flavus and on Bacillus thuringiensis RSKK 380; B. cereus M10 on Bacillus megaterium RSKK 578, on Bacillus thuringiensis RSKK 380 and on Bacillus cereus F2; B. brevis M4 and B. cereus M5 on Bacillus cereus F2. Also, the inhibition effect of isolates between each others were carried by using agar diffusion method and the inhibition effect some isolates were found. Proteolytic activities of Bacillus isolates were tested in Skim Milk Agar (SMA). It was concluded that all strains had proteolytic activities except for B. sphaericus M3 and B. sphaericus ATCC 14577. Proteolytic activities of the isolates was found between 1,4-8,1 mm zone diameter. The overage of proteolytic activity was found to be 4,17 mm. The highest zone diameter of proteolysis 8,1 mm was found in B. circulans M32 strain. Plasmid DNA profiles of Bacillus strains were determined and their moleculer weights were found. Results indicated that only six strains were carried plasmid DNA and their moleculer weights were ranged from 3,9 to 3,1 kb.VI Science code : 216 Key Words : Bacillus, isolation, protein profile, bioplastic (PHB) production, antimicrobial activity, proteolytic activity, plasmid DNA.