O-۴۶ Screening and mutagenesis of the potential microbes to enhance the conversion of Poultry Slaughterhouse

Background and Aim: Due to rapid urbanization and changes in consumer behavior, the abundance of solid waste produced became a new challenge to most of the developing countries. The common biological treatments available are composting and fermentation, which turned organic wastes into valuable products. Detection microbes by good protease activity is an economic way for degradation Poultry Slaughterhouse.Methods: In this study ۲۸ bacterial strains isolated from Poultry Slaughterhouse and soil were screened for their ability to produce proteases. Isolation of bacteria was done using serial dilution method on skim milk agar. All isolated bacteria were screened for their ability to produce protease enzyme. The bacterial isolate showing maximum alkaline protease activity was identified using ۱۶ S rRNA genetic identification. To induce mutations, ultraviolet (UV) irradiation was used. The most active mutant strains were selected for production, purification, and characterization of alkaline protease followed by evaluation of alkaline protease activity Results: According to the biochemical and morphological characters, the isolated strain was grouped under the genus of Staphylococcus, lactobacillus and enterobacteriaceae. Based on ۱۶S rRNA typing and phylogenetic analysis four isolated were identified as and according to diameter clear zone on keratin agar medium, exhibited higher level of keratinolytic activity among other isolated strains. Taken together, these results suggest new keratinase producing bacteria with high keratinolytic activities which have the potential use in industrial biotechnology. The cell extracts of four most potent producers were further examined and it was found that their proteases exhibited maximum activity at range of pH ۶.۸–۸.۱. Gelatin zymography revealed that two of the selected gram positive strains produced multiple active SDS-resistant proteases. On the other hand, PCR amplification of protease gene sequences of total DNA from all isolates yielded four distinct amplification fragments of ۶۵۰, ۴۵۰ and ۳۰۰ bp, which might have been derived from different serine protease genes. Besides it was done by four bacteria by combined mutagenesis using UV, these strains were further improved with regard to their alkaline protease production. The alkaline protease activity of the mutant strain BP۲۳ was greatly improved up to ۸۰۰۰ U/ml, in comparison with its parent strain of ۱۰۰۰ U/ml.Conclusion: Detection microbes by good protease activity is an economic way for degradation Poultry Slaughterhouse.

کلیدواژه ها:

proteases ، Poultry Slaughterhouse ، PCR ، fermentation