ABSTRACT

High collagenolytic activity has been detected in pathogenic bacteria. Collagenase plays an essential role in the invasion step in animals and humans. In this study, we characterized collagenase found in the nonpathogenic bacterium Lysinibacillus sphaericus VN3, which was isolated from soil in Vietnam. The collagenase activity of the purified enzyme was strongly inhibited by Cu²⁺, but it was significantly increased by Zn²⁺. The purified enzyme with a molecular mass of approximately 110 kDa exhibited collagenolytic, gelatinolytic, and caseinolytic activity. The kinetic studies showed that this enzyme had greater hydrolyzing activity toward collagen and gelatin compared with casein. Based on the ratio V _max/K _m, collagen is likely to be the best substrate among three proteins. We found that this collagenase could digest small pieces of bovine skin and tendon into a collagen solution. Interestingly, at pH 6.0–8.0, the soluble collagen could form a collagen membrane, which is useful as a wound-healing biomaterial.

摘要

在致病细菌中已检测到高的胶原分解活性。胶原酶在动物和人类的入侵步骤中起着至关重要的作用。在这项研究中，我们表征了从越南土壤中分离出的非致病性球形芽孢杆菌VN3中发现的胶原酶。纯化的酶的胶原酶活性受到Cu ^2+的强烈抑制，但被Zn ²⁺显着提高。分子量约为110 kDa的纯化酶具有胶原分解，明胶分解和酪蛋白分解活性。动力学研究表明，与酪蛋白相比，该酶对胶原和明胶具有更大的水解活性。基于比率V _max / K _m，胶原蛋白可能是三种蛋白质中最好的底物。我们发现这种胶原酶可以将小块的牛皮肤和肌腱消化成胶原溶液。有趣的是，在pH 6.0-8.0时，可溶性胶原蛋白可以形成胶原蛋白膜，可用作伤口愈合的生物材料。