RESUMO
The aim of this study was to investigate the antibacterial interactions of pulegone and 1,8-cineole with monolaurin ornisin against Staphylococcus aureus. The individual and combined antibacterial activities of the compounds were evaluated using minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), fractional inhibitory concentration index (FICi), and time-kill methods. Furthermore, the mechanism of the antibacterial action of the compounds was tested by measuring the release of cell constituents. The MIC values of pulegone, 1,8-cineole, nisin, and monolaurin were 5.85 µl/ml, 23.43 µl/ml, 6.25 µg/ml, and 0.031 mg/ml, respectively. A synergistic antibacterial activity (FICi = 0.5) was found between 1,8-cineole and nisin. The time-kill assay showed that the populations of S. aureus exposed to 1,8-cineole, nisin, and their combination were decreased by 5.9, 5.3, and 7.1 log CFU (colony-forming units)/mL, respectively. The combination of 1,8-cineole and nisin also induced the highest release of cell constituents. It was concluded that the combination of 1,8-cineole and nisin could be considered as a novel and promising combination which may reduce the required dose of each antibacterial compound.
RESUMO
BACKGROUND AND OBJECTIVES: Proteases are a group of enzymes that catalyze the degradation of proteins resulting in the production of their amino acid constituents. They are the most important group of industrial enzymes which account for about 60% of total enzymes in the market and produced mainly by microorganisms. The attempts were made to study the kinetic parameters of protease produced by Streptomyces griseoflavus PTCC1130. MATERIALS AND METHODS: Streptomyces griseoflavus PTCC1130 was grown on casein agar. Different media such as BM1, BM2, BM3 and BM4 were prepared. Data obtained from growth and protease production were subjected to kinetics evaluation. Casein was used as substrate for protease activity and the released soluble peptide bearing aromatic amino acid were quantified by Folin Cioclateaue reagent. Protein content of the enzyme and the sugar utilized by the organism were estimated by Bradford and Miller's methods respectively. RESULTS: Basal Medium named as BM1, BM2, BM3 and BM4(50 mL in 250 mL Erlen Meyer flasks) were screened out to evaluate protease production by Streptomyces griseoflavus PTCC1130. They were inoculated with known amount of seed culture and kept on rotary shaker. To obtain the specific growth rate, wet weight of biomass was plotted against the time. The clarified supernatant was used for the analysis of protease by measuring the soluble peptide containing aromatic amino acid residues employing Folin Cioclateaue reagent. Our results showed that maximum level of enzyme production (14035 U/L) was occurred at late exponential phase using Basal Medium supplemented with zinc sulfate (0.5g/L), casein (10g/L) at pH 6.5. CONCLUSIONS: A kinetic study of protease production by Streptomyces griseoflavus PTCC1130 provided highly quantitative information regarding the behavior of a system, which is essential to study the fermentation process. Exploitation of such kinetics analysis would be useful in commercialization of microbial enzyme production.
