Enhancing the enzyme production from alkaline and thermostable keratinase producer native Bacillus sp. and Streptomyces sp. strains by genetic manipulation, and enzyme characterization

Alkaline keratinase are widely used in several fields including leather processing, detergents, agricultural-animal industry, and production of protein hydrolysates, etc. Although, there are many researches on microbial alkalin keratinase, the studying about enhanced enzyme production with low cost substrat is still continues intensely. In this study, natural Bacillus sp. and Streptomyces sp. strains which are the producer of alkali and thermo-stabile keratinase were isolated, identified and developed. Also, medium conditions, biochemical characterization of enzymes and their industrial usability was analyzed. In mutation studies was detected that while the reproduction time of UV mutant C11-UV8 strain decreased, the enzyme yield increased 2.7 times (1405 U/ml), and keratinase production of the EMS mutant C1-EMS80 strain increased 8 times and is very stable compared to the main strain at wide temperature range. Bacillus licheniformis C11 keratinaze gen was cloned into pET22b(+) vector system and transferred to the E.coli (Rosetta-gami (DE3)pLysS) cell. Recombinant protein (keratinase) production showed an increase of 3.3 times compared to the main strain at the end of 18 hours. According to the obtained data, native and recombinant keratinase enzyme can be used in several industry area including leather, detergents, bioremiation, feed and agriculture etc.