立足于目前制革行业的清洁化发展需求，为了有效解决目前脱毛过程中存在的固体废弃物污染问题，本项目以脱毛酶作为研究对象，针对脱毛过程中“提高脱毛酶专一性作用机制”的基础科学问题，使用基因工程手段从全新的角度提出了解决方案。在研究过程中，首先从具有高产角蛋白酶特性的解淀粉芽孢杆菌中提取角蛋白酶基因，对其进行定点突变，提高其催化活性与稳定性，然后将其在缺陷8种主要胞外蛋白酶的枯草芽孢杆菌WB600中进行异源表达，从而去除传统脱毛酶中存在的对胶原蛋白结构有破坏作用的酶组分；在此基础上优化发酵条件，得到对胶原无降解作用的新型脱毛酶，并对新型脱毛酶的作用机制进行总结；最后，阐述脱毛过程必须存在的金属离子Ca2+/Na+对脱毛酶作用的影响机制及脱毛体系皮-脱毛酶-Ca2+/Na2+之间的构效关系，为脱毛酶的研究和开发以及制革行业的革新奠定了理论基础。

Unhairing process brought serious pollution, and enzyme application for replacing polluting chemicals in unhairing process attracted much attentions in recent years. However, the unhairing enzymes haven’t been accepted widely in actual production due to low purity,complex composition and poor stability. To solve these problems, unhairing enzyme is suggested to be improved by genetic modification in this project. High-keratinase-producing gene will be extracted from Bacillus Amyloliquefaciens strain, and transferred into Bacillus subtilis WB800 to construct recombinant strain, which produces unhairing enzyme without collagenase. Then the properties of modified unhairing enzyme will be analyzed and optimized. Furthermore, how modified unhairing enzyme act on unhairing process will be further studied. In addition, the relationship among the hide, modified unhairing enzyme and Ca2+/Na+ will be elucidated, while the activating effect, affecting factors and regulation method will be investigated. This research will promote the application of unhairing enzyme in leather production.