Marine Microorganisms Source Collagenase Enzyme System Research

As one of the most important proteins in vivo, collagen mainly exists in the bones and skin. Only after being decomposed into peptides and amino acids can it be assimilated by the body. Therefore, enzymatically decomposing collagen into collagen peptides will enhance the assimilation of such protein. The collagenase, which contributes to such assimilation, has been obtained from tissues and microorganismsThe marine mixed bacteria are used as the main subject of the study. Firstly, a strain of highly active collagen enzyme-producing bacteria was sifted out. Secondly, analysis was made of properties of the enzyme system in such crude enzyme. Thirdly, using collagenase as the principal product, comprehensive optimization was made of the bacteria’s nutrient media components and fermentation conditions to get the best enzyme-producing conditions, thus providing a measure of technical reference to the industrialization production of collagen peptides.After the preliminary and secondary screening of marine mixed bacteria, a strain of bacteria with a high collagen enzyme activity was obtained. Then, through fishskin digestion trial, it showed that collagen is highly decomposable in the presence of such bacteria. Single factor way was used to analyze the enzyme system in such crude enzyme. Finally, discussion was made about the influence of pH, thermal stability of the enzyme, temperature and metal ions on the activity of collagenase, lipase, amylase and cellulose, respectively.The results showed that:the enzyme system of the bacteria contains a relatively high level of collagenase and lipase but an extremely low level of amylase and cellulose. Collagenase activity peaks at pH 8.0 as 57.6 U/ml; lipase activity peaks at pH 8.3 as 29.4 U/ml. Taking the thermal stability and temperature into the final analysis, when the temperature is 40℃, Mg2+, Cu2+, Fe2+ and Mn2+ show little positive influence on collagenase activity, while Mg2+ and Mn2+ are conducive to lipase activity, by the increase rate of 24.5% and 14.1%, respectively. Through the experiment, the optimum condition for the bacteria to produce collagenase is obtained:glucose 3 g/L, yeast extract powder 9 g/L, NaCl 5 g/L, MgCl2·6H2O 4 mg/L, KCl 5 mg/L and pH 8.0; the culture time is 40 h; the temperature is 30℃; well-oxygenated.Under such optimum condition, collagenase activity was increased by 54%. After the re-measurement of lipase, amylase and cellulose in the fermentation liquor, it turned out that lipase was 44 U/ml, equivalent to that from other common lipase-producing bacteria. While amylase and cellulose were little found. Therefore, the bacteria can also be made use of in the development and production of lipase-producing bacteria.