生物技术通报 ›› 2017, Vol. 33 ›› Issue (5): 183-189.doi: 10.13560/j.cnki.biotech.bull.1985.2016-1098

• 研究报告 • 上一篇    下一篇

一种新型蛋白酶的酶学特性及脱毛应用

郑翔1, 杨何宝1, 胡美荣2, 刘春卯1, 吴芳彤1, 曹倩荣1   

  1. 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
  • 收稿日期:2016-12-05 出版日期:2017-05-25 发布日期:2017-05-19
  • 作者简介:郑翔,男,硕士,研究方向:酶制剂的开发与应用;E-mail:569186912@163.com
  • 基金资助:
    河北省省级省校科技合作开发资金项目(2016049819)

Enzymatic Characteristics of a Novel Protease and Its Application for Dehairing

ZHENG Xiang1, YANG He-bao1, HU Mei-rong2, LIU Chun-mao1, WU Fang-tong1, CAO Qian-rong1   

  1. 1. Enzyme Preparation Office,Institute of Microbiology,Hebei Academy of Sciences,Baoding 071051; 2. CAS Key Laboratory of Microbial Physiological and Metabolic Engineering,Institute of Microbiology,Chinese Academy of Sciences,Beijing 100010
  • Received:2016-12-05 Published:2017-05-25 Online:2017-05-19

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/pHT43-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;Ca2+、Mn2+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。

关键词: 制革业, 清洁化技术, 蛋白酶, 酶学性质

Abstract: 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 alternative to the conventional chemical process.

Key words: leather manufacture, clean-technology, protease, enzymatic properties