Isolation And Identification And Culture Optimization Of Microorganism For The Collagenase Production

Collagen has a unique super-helical structure which refers to three α helix peptide chain of the right hand. General processing temperature and short heating do not make collagen breakdown. Stable nature of collagen which making it difficult to digest is not easily absorbed by the body. Collagen can be hydrolyzed collagen peptides. Collagen peptides have easily absorbed, anti-ulcer, anti-allergy, blood pressure, anti-bacterial, anti-oxidation,cholesterol-lowering, anti-aging, promote wound healing, prevention of osteoporosis and arthritis, promote corneal epithelial wound healing and other physiological activity. These physiologically active peptides of collagen makes collagen become efficient utilization of research focus. This article aims to explore the hydrolyzed collagen can get by way of microorganisms high activity of collagenase. Finally, in order to achieve the purpose of the factory production, collagenase production process has been optimizated.First of all,the microorganism for collagenase production was isolated from the offshore in Yantai, and obtained the strain SM12. It was belonged to Staphylococcus succinus with the method of morphologic observation and molecular biology. Optimal culture conditions and components are 20 g / L glucose, 15 g / L beef extract, 20 g / L NaCl, pH7.5, the culture temperature 37 ℃, inoculum 7.5%(v / v) and liquid volume 100 mL / 500 mL bottle. The highest activity of collagenase from Staphylococcus succinus SM12 was 185.32U/mL at 24 h,with optimization of culture components and conditions. Visiblely, strain SM12 under optimal conditions producing the enzyme activity is still not high.This study focuses on breeding innovation strain.Due within Japan hippocampus intestinal screened antagonistic bacteria strain(No. HS17) can compete in the same space or nutritional niche, the bacteria secrete antibiotics or other factors such as inhibiting the growth of microorganisms and demonstrated strong vitality, we can find that HS17 has the potential to yield high activity of collagenase. We have tried to study collagenase production, in order to obtain a greater yield than strains SM12 collagenase. It was belonged to Bacilus sp. with the method of morphologic observation and molecular biology. Optimal culture conditions and components are 3 g / L of gelatin, 50 g / L of fructose, 4 g / L yeast extract, 4 g / L ZnSO4, 3 g/ L MnSO4, 3 g / L FePO4.2H2 O, 6 g / L(NH4) 2SO4, 4 g / L Na2HPO4, 5 g / L K2HPO4,inoculation 4.46%(v / v), culture temperature 30 ℃ and medium initial pH 7. Dissolved oxygen(DO) is maintained at least 5% in the fermentation process controls by stirring speed and aeration rate. The highest activity of collagenase from Staphylococcus succinus SM12 was 415.39U/mL at 35 h, with optimization of culture components and conditions. In the late fermentation, with the gradual decrease of cell concentration, collagenase activity also showed a gradual decrease. Thus, Bacillus sp. HS17 which is easy to cultivate,nonpathogenic and efficient degradation, having a broader application prospects and practical significance.