一种新型蛋白酶的酶学特性及脱毛应用  

Enzymatic Characteristics of a Novel Protease and Its Application for Dehairing

作　　者：郑翔 杨何宝 胡美荣[2] 刘春卯 吴芳彤 曹倩荣 

机构地区：[1]河北省微生物研究所酶工程室,保定071051 [2]中国科学院微生物研究所微生物生理与代谢工程重点实验室,北京100101

出　　处：《生物技术通报》2017年第5期183-189,共7页Biotechnology Bulletin

基　　金：河北省省级省校科技合作开发资金项目(2016049819)

摘　　要：蛋白酶可作为清洁化产品应用在制革脱毛工艺,旨在明确枯草工程菌株WB800N/p HT43-npr发酵生产重组中性蛋白酶(JW-3蛋白酶)的脱毛效果及脱毛工艺条件,为清洁化脱毛制革工艺提供技术参考。研究了JW-3蛋白酶的热稳定性、pH耐受性,金属离子及化学试剂对酶活力的影响,酶解胶原蛋白的能力,同时对酶法与传统灰碱法黄牛皮脱毛效果进行评价。结果显示,JW-3蛋白酶的耐高温能力差,40℃保存1 h酶活力便开始衰减;在pH6、pH7稳定性良好;Ca^(2+)、Mn^(2+)对酶活力有激活作用;在表面活性剂Triton X-100、Tween20、Tween80中稳定;JW-3蛋白酶不具有水解胶原蛋白的能力,与传统灰碱法脱毛的皮粒面效果相当,且可以回收完整的动物毛发再利用。JW-3酶法可替代灰碱法脱毛实现清洁化脱毛制革工艺。Protease can be applied in dehairing process of leather industry as an eco-friendly product. We aim to clear the dehairing performance and the process of enzymatic dehairing of JW-3 protease produced by recombinant strain WB800N/pHT43-npr, and to provide technical support for clean technology of leather manufacture. The thermostability and pH value tolerance of JW-3, as well as the effects of metal ions and chemicals on its enzyme activity were investigated. Meanwhile, the ability of enzymatic hydrolysis on collagen, and the comparison between enzymatic dehairing and lime-sulfide dehairing process on cowhide were evaluated. Results showed that JW-3 protease was in poor thermal resistance that enzyme activity began to decay after remaining 1 h at 40℃, and presented solid stability at pH 6 and 7, and was stable in the surfactant of Triton X-100, Tween20, Tween80 ; moreover, metal ions of Ca2＋, Mn2＋ activated its activity. Furthermore, JW-3 protease was incapable of hydrolyzing collagen, and showed the equivalent dehairing performance with lime-sulfide process, also by which nondestructive depilated hair could be recycled. Thus, JW-3 protease can complete cowhide hair removal process, but not hydrolyze the collagen of cowhide, and can be used as an environmentally friendly ahernative to the conventional chemical process.

关 键 词：制革业 清洁化技术 蛋白酶 酶学性质 

分 类 号：Q814.9[生物学—生物工程] TS529[轻工技术与工程—皮革化学与工程]