Amylases are one of the most important enzymes in present-day biotechnology. The present study was concerned with the production and partial characterization of extracellular amylase from Bacillus amyloliquefaciens P-001. The effect of various fermentation conditions on amylase production through shake-flask culture was investigated. Enzyme production was induced by a variety of starchy substrate but corn flour was found to be a suitable natural source for maximum production. Tryptone and ammonium nitrate (0.2%) as nitrogen sources gave higher yield compared to other nitrogen sources. Maximum enzyme production was obtained after 48 hrs of incubation in a fermentation medium with initial pH 9.0 at 42°C under continuous agitation at 150 rpm. The size of inoculum was also optimized which was found to be 1% (v/v). Enzyme production was 2.43 times higher after optimizing the production conditions as compared to the basal media. Studies on crude amylase revealed that optimum pH, temperature and reaction time of enzyme activity was 6.5, 60°C and 40 minutes respectively. About 73% of the activity retained after heating the crude enzyme solution at 50°C for 30 min. The enzyme was activated by Ca2+ (relative activity 146.25%). It was strongly inhibited by Mn2+, Zn2+ and Cu2+, but less affected by Mg2+ and Fe2+.

Background and Purpose: Klebsiella pneumoniae and Klebsiella oxytoca are the two most common pathogens causing nosocomial infections in humans and are of great concern for developing multidrug resistance. In the present study, K. pneumoniae and K. oxytoca from clinical samples were evaluated for their antibiotic sensitivity patterns against commonly used antibiotics and production of extended-spectrum beta-lactamase (ESBL). Ampicillin, Amoxicillin, Ceftriaxone, Ciprofloxacin, Gentamicin, Nalidixic acid, Tetracycline was 100%, 90%, 45%, 40%, 45%, 25%, 50%, 35% respectively. Multidrug resistance was found more common in K. pneumoniae (56%) than in K. oxytoca (50%). Prevalence rate of ESBL producing Klebsiella was found 45% among which K. pneumoniae (50%) were found more prominent than K. oxytoca (25%). All the ESBL producing Klebsiella isolates were found to be multidrug resistant, showing 100% resistance to Ampicillin, Amoxicillin, Ceftriaxone and Ciprofloxacin. Materials and Methods:

Proteolytic enzymes have occupied a pivotal position for their practical applications. The present study was carried out under shake flask conditions for the production of alkaline protease from Bacillus licheniformis P003 in basal medium containing glucose, peptone, K2HPO4, MgSO4 and Na2CO3 at pH 10. The effect of culture conditions and medium components for maximum production of alkaline protease was investigated using one factor constant at a time method along with its characterization. Maximum level of enzyme production was obtained after 48h of incubation with 2% inoculum size at 42°C, under continuous agitation at 150 rpm, in growth medium of pH 9. Highest enzyme production was obtained using 1% rice flour as carbon source and 0.8% beef extract as organic nitrogen source. Results indicated that single organic nitrogen source alone was more suitable than using in combinations and there was no significant positive effect of adding inorganic nitrogen sources in basal medium. After optimization of the parameters, enzyme production was increased about 20 fold than that of in basal medium. The crude enzyme was highly active at pH 10 and stable from pH 7–11. The enzyme showed highest activity (100%) at 50°C, and retained 78% relative activity at 70°C. Stability studies showed that the enzyme retained 75% of its initial activity after heating at 60°C for 1h. The enzyme retained about 66% and 46% of its initial activity after 28 days of storage at 4°C and room temperature (25°C) respectively. Mn2+ and Mg2+ increased the residual activity of the enzyme, whereas Fe2+ moderately inhibited its residual activity. When pre-incubated with Tween-20, Tween-80, SDS and H2O2, each at 0.5% concentration, the enzyme showed increased residual activity. These characteristics may make the enzyme suitable for several industrial applications, especially in leather industries.

Background and Purpose: Klebsiella pneumoniae and Klebsiella oxytoca are the two most common pathogens causing nosocomial infections in humans and are of great concern for developing multidrug resistance. In the present study, K. pneumoniae and K. oxytoca from clinical samples were evaluated for their antibiotic sensitivity patterns against commonly used antibiotics and production of extended-spectrum beta-lactamase (ESBL). Materials and Methods: The isolates were obtained from tracheal swabs, sputum, wound swabs, pus, blood and urine samples of hospitalized patients. Klebsiella pneumoniae and Klebsiella oxytoca were identified by cultural and biochemical methods. Antibiotic sensitivity test was performed by modified Kirby-Bauer disc diffusion technique. ESBL production in Klebsiella spp. was confirmed by double disc synergy test.Results and Conclusion: Out of 500 clinical isolates, 120 were found positive for Klebsiella among which 108 were K. pneumoniae and 12 were K. oxytoca based on indole test. Prevalence rate of Klebsiella was found more prominent in males aged over 50 years, mostly in urine samples. Overall resistance pattern of Klebsiella isolates to Ampicillin, Amoxicillin, Ceftriaxone, Ciprofloxacin, Co-trimoxazole, Gentamicin, Nalidixic acid, Tetracycline was 100%, 90%, 45%, 40%, 45%, 25%, 50%, 35% respectively. Multidrug resistance was found more common in K. pneumoniae (56%) than in K. oxytoca (50%). Prevalence rate of ESBL producing Klebsiella was found 45% among which K. pneumoniae (50%) were found more prominent than K. oxytoca (25%). All the ESBL producing Klebsiella isolates were found to be multidrug resistant, showing 100% resistance to Ampicillin, Amoxicillin, Ceftriaxone and Ciprofloxacin. Keywords: Multi-drug resistant, Klebsiella, extended-spectrum beta-lactamase.

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