摘要
旨在优化菌株Bacillus sp.50-3产角蛋白酶的发酵工艺及纯化。研究不同发酵条件对菌株Bacillus sp.50-3产角蛋白酶的影响,采用硫酸铵沉淀和阴离子交换柱层析对酶进行分离纯化。结果:角蛋白酶最佳发酵条件为60 mL/300 mL、pH=8.0、种龄12 h、接种量2%、温度37℃、140 r/min摇床培养36 h;40%~80%的硫酸铵沉淀可以去除发酵酶液中的大部分杂蛋白,通过DEAE-Sephadex-A50阴离子交换柱层析,经SDS-PAGE,考马斯亮蓝染色得到一条电泳条带,分子量约为30 kDa,经酶活性电泳检测具有角蛋白酶活力。综上,该研究结果对角蛋白酶在工业生产中的推广应用具有重要的参考价值。
This experiment aimed to optimize the fermentation process of keratinase production by the Bacillus sp.50-3 strain and to purify the enzyme.Different fermentation conditions were studied for effects of keratinase from the strain Bacillus sp.50-3.Ammonium sulfate pre-cipitation and anion-exchange column chromatography were used to separate and purify the keratinase.The results showed that the optimal fermentation conditions were 60 mL/300 mL,pH=8.0,seed age 12 h,inoculation amount 2%,temperature 37℃,and 140 r/min shaker culture for 36 h.The keratianse was purified through 40%-80%of(NH_(4))_(2)SO_(4)precipitation and DEAE-Sephadex-A50 ion exchange col-umn.It was found that the co-omassie brilliant blue R-250 stain obtained one band with keratinase activity determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.Its molecular weight was about 30 kDa.In summary,this study is of important reference signifi-cance for the application of keratinase in industrial production.
作者
岳晓禹
崔丽伟
常惟丹
李长滨
李欣
许文涛
YUE Xiaoyu;CUI Liwei;CHANG Weidan;LI Changbin;LI Xin;XU Wentao(College of Food and Biotechnology Engineering,Henan University of Animal Husbandry and Economy,Zhengzhou 450046,China;Department of Nutrition and Health,China Agricultural University,Beijing 100083,China)
出处
《畜牧与兽医》
北大核心
2025年第12期27-33,共7页
Animal Husbandry & Veterinary Medicine
基金
河南省科技发展计划项目(232102111068)
河南省高等学校重点科研项目计划项目(22A550012)。
关键词
芽胞杆菌
角蛋白酶
发酵条件
纯化
Bacillus
keratinase
fermentation conditions
purification
