地衣芽孢杆菌-角蛋白酶联合降解羽毛体系的优化及产物应用  

Optimization and product application of feather-degrading system in combination of Bacillus licheniformis and keratinase

作　　者：张宝玉 彭政 张娟 李江华 ZHANG Baoyu;PENG Zheng;ZHANG Juan;LI Jianghua(School of Biotechnology,Jiangnan University;Key Laboratory of Industrial Biotechnology,Ministry of Education/Science Center for Future Foods,Jiangnan University;Engineering Research Center of Food Synthetic Biotechnology,Ministry of Education,Jiangnan University,Wuxi,Jiangsu 214122,China)

机构地区：[1]江南大学生物工程学院 [2]江南大学工业生物技术教育部重点实验室/未来食品科学中心 [3]江南大学食品合成生物技术教育部工程研究中心,江苏无锡214122

出　　处：《福建农林大学学报（自然科学版）》2022年第6期827-834,共8页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：江苏省重点研发(社会发展)项目(BE2021624)。

摘　　要：为了提高羽毛降解率,本研究拟开发一种重组角蛋白酶KerZ1和地衣芽孢杆菌(Bacillus licheniformis)BBE11-1联合降解羽毛的体系.通过单因素和正交试验设计对B.licheniformis BBE11-1培养体系及其与角蛋白酶KerZ1的复配条件进行优化,并利用氨基酸分析仪测定水解液中的氨基酸含量.在温度40℃、pH 10、角蛋白酶活性50 kU·mL^(-1)、B.licheniformis BBE11-1接种量10%的条件下水解36 h,100 g·L^(-1)鸭毛的降解率达74.0%,分别较KerZ1和B.licheniformis BBE11-1单独降解提高了18.0%和15.0%,且水解液中总氨基酸含量为7472.33 mg·L^(-1);在温度40℃、pH 10、角蛋白酶活性30 kU·mL^(-1)、B.licheniformis BBE11-1接种量8%的条件下水解32 h,100 g·L^(-1)鸡毛的降解率为64.0%,分别比KerZ1和B.licheniformis BBE11-1单独降解提高了29.0%和28.5%,水解液中总氨基酸含量为5589.67 mg·L^(-1).另外,将水解液添加到培养基中可以显著提高菌株WB600-pP43NMK-ker的菌体密度和角蛋白酶活性.因此,B.licheniformis BBE11-1和角蛋白酶KerZ1联合可以高效降解羽毛,且羽毛水解液在微生物培养中极具应用潜力.A feather-degrading system which combined recombinant keratinase KerZ1 and Bacillus licheniformis BBE11-1 was developed to improve feather degradation efficiency.The B.licheniformis BBE11-1 culture system and complex system supplemented with keratinase KerZ1 were optimized via single factor experiment and orthogonal experimental design,and the amino acid content of the hydrolysate was determined using amino acid analyzer.The results showed that the degradation rate of 100 g·L^(-1) duck feather reached 74.0%when the system was inoculated with 10%B.licheniformis BBE11-1 and 50 kU·mL^(-1) keratinase for 36 h at 40℃and pH 10,resulting in a total amino acid content of 7472.33 mg·L^(-1) in the hydrolysate,whose degradation rate was 18.0%and 15.0%higher than that inoculated with KerZ1 or B.licheniformis BBE11-1 alone,respectively.Similarly,the degradation rate reached 64.0%when inoculated with 8%B.licheniformis BBE11-1 and 30 kU·mL^(-1) keratinase for 32 h at 40℃and pH 10,and the degradation rate was increased by 29.0%and 28.5%than that inoculated with KerZ1 or B.licheniformis BBE11-1 alone,respectively,yielding a total amino acid content of 5589.67 mg·L^(-1) in the hydrolysate.Moreover,the addition of the hydrolysate into culture medium increased the cell density of WB600-pP43NMK-ker and keratinase activity significantly.To summarize,the feather-degradation system in combination of B.licheniformis BBE11-1 and keratinase KerZ1 was efficient,and its hydrolysate was of great potential in microbial culture.

关 键 词：废弃羽毛 地衣芽孢杆菌 角蛋白酶 协同作用 降解率 氨基酸 

分 类 号：Q939.9[生物学—微生物学]