SCREENING AND MOLECULAR CHARACTERIZATION OF BACILLUS SP PR01 FOR THE PRODUCTION OF PROTEASE ENZYME
The current study was attempted to isolate and characterize the protease producing Bacillus sp. Among the 3 strains isolated from soil sample collected f rom kuppam region, Chennai, India. One strain (PR01) exhibited potent proteolytic activity. Based on the morphological, biochemical and molecular characterization, the isolate (PR01) was identified as Bacillus sp. The strain was designated as Bacillus sp P R01. The maximum growth of Bacillus sp PR01was observed at pH 7. The optimum temperature for the growth of Bacillus sp PR01 was found to be 27 ° C an d incubation time for 2 days. The maximum growth was noted when yeast extract was used as a nitrogen source a nd dextrose was used as the carbon source. The production of enzyme Bacillus sp PR01 was carried out in production medium. The t otal protein concentration was determined and the specific activity was also performed by casein hydrolysis assay.
Microbial proteases: Detection, production, and genetic improvement
Microbial proteases are one of the important groups of industrially and commercially produced enzymes contributing approximately 2/3 of all enzyme sales. Though proteases are produced by many microorganisms, emphasis is on the microorganisms producing proteases with desired characters. As demand for novel proteases is increasing day by day the initial screening methods and assays for protease detection are of utmost importance. This review focuses attention on present status of knowledge on the various methods and protocols available for protease screening, detection, and quantification starting from plate assays to spectrophotometric, fluorometric, and nanoparticles based assays. The review will help in making strategies for exploitation of protease resources and improvement of enzymes to obtain more robust proteases.
Detection of extracellular proteinase of Pseudomonas fragi by enzyme-linked immunosorbent assay
A double-antibody-sandwich, enzyme-linked immunosorbent assay was developed to detect an extracellular proteinase produced by Pseudomonas fragi. The method was capable of detecting 4 μg/ml of the proteinase in spiked samples of buffer and broth and 4.2 μg/ml in a broth culture of the organism. The assay detected the presence of proteinase at bacterial densities of approximately 104 cfu/ml, which develop after incubation for 15 h at 25°C in a broth medium. All assays could be completed within 7 h. This assay is of value in plotting proteolytic expression in relation to the growth cycle of Ps. fragi in broth culture and may be of value, with development, in other more complex milieux.