角蛋白酶kerC基因的克隆及序列分析

发布时间：2018-04-12 07:37

本文选题：角蛋白酶 + 枯草芽孢杆菌　；参考：《基因组学与应用生物学》2017年05期

【摘要】：为克隆分析角蛋白酶ker C基因,本研究以羽毛为底物从11株实验室保存的芽孢杆菌中筛选角蛋白酶产生菌。得到了一株具有高效降解羽毛能力的枯草芽孢杆菌BS10,该菌株3 d即可降解一根完整的羽毛,以羽毛粉、天青角蛋白为底物测定其酶活力,分别达(1.88±0.10)U/mL、(1.79±0.49)U/mL。以同源克隆的方法克隆ker C基因,获得了一条全长1 149 bp的kerC基因(Gen Bank登录号:KX108888),编码383个氨基酸。利用BLAST、ProtParm、SOPMA和MEGA等生物信息学工具对其理化性质、二级结构及系统进化树等进行分析,发现其与NCBI中的角蛋白酶基因相似性达到85%;相对分子质量为39.095 kD,等电点为9.23;该基因蛋白为亲水性蛋白;系统进化树分析结果与菌株信息相吻合。羽毛降解菌的获得可促进废弃羽毛资源化利用;角蛋白酶基因的获得及其分子特征的分析,为进一步通过基因工程手段提高角蛋白酶活性提供了一定的理论依据。
[Abstract]:In order to clone and analyze the ker C gene of keratinase, keratinase producing bacteria were screened from 11 strains of Bacillus in laboratory with feather as substrate.A strain of Bacillus subtilis BS10 was obtained, which could degrade a whole feather in 3 days. The enzyme activity of BS10 was determined with feather powder and azure keratin as substrate. The enzyme activity was 1.88 卤0.10 U / m L ~ + 1.79 卤0.49 U / m ~ (-1) 路L ~ (-1), respectively.Ker C gene was cloned by homologous cloning, and a 1 149 BP kerC gene named Gen Bank was obtained, which encodes 383 amino acids.The physicochemical properties, secondary structure and phylogenetic tree were analyzed by using bioinformatics tools such as BLASTT-ProtParm-SOPMA and MEGA.It was found that the similarity between the protein and the keratinase gene in NCBI was 85%; the relative molecular weight was 39.095 KD and the isoelectric point was 9.23; the protein was hydrophilic protein; the phylogenetic tree analysis results were consistent with the information of the strain.The acquisition of feather degrading bacteria can promote the utilization of waste feathers, and the acquisition of keratinase gene and the analysis of its molecular characteristics provide a theoretical basis for further enhancing the activity of keratinase through genetic engineering.
【作者单位】：四川农业大学生命科学学院;
【分类号】：Q55;Q78

【相似文献】