干旱区生态环境知识资源中心(Arid): 戈壁异常球菌角蛋白酶基因的克隆及功能研究

Arid

	戈壁异常球菌角蛋白酶基因的克隆及功能研究
其他题名	Cloning and Identification of Keratinase Gene from Deinococcus gobiensis I-0
	耿秀秀 1; 周正富 2; 刘盈盈 2; 平淑珍 2; 王劲 1
来源期刊	生物技术通报
ISSN	1002-5464
出版年	2019
卷号	35 期号:3 页码:65-70
中文摘要	旨为深入发掘角蛋白酶重要的基因资源,进一步提高其水解酶活,为工业生产提供理论基础。从分离于新疆戈壁沙漠的戈壁异常球菌(Deinococcus gobiensis)I-0中克隆出一个编码角蛋白酶的基因并命名Dgker。构建原核表达载体pET -22b -Dgker并进行体外诱导表达纯化,通过对羽毛水解活性的测定探究粗酶液的最适温度和pH。结果显示,Dgker蛋白N端前50个氨基酸对蛋白的表达纯化有很大影响。Dgker的重组菌株3d完全分解羽毛,在奶粉平板上重组菌株粗酶液比空载菌株粗酶液降解圈更显著。Dgker适应的温度和pH较为广泛其中最适温度是60℃、最适pH是5.0。Dgker能降解羽毛在以后的工业生产及处理废弃羽毛有极大的应用空间。
英文摘要	In order to further tap the important gene resources of keratinase,improve its hydrolytic activity,and provide theoretical basis for industrial production,this experiment cloned a gene encoding keratinase from Deinococcus gobiensis I-0 isolated from Gobi desert of Xinjiang and named it as Dgker. Prokaryotic expression vector pET-22B-Dgker was constructed and then induced,expressed and purified in vitro,the optimal temperature and pH of the crude enzyme solution were determined through the hydrolysis activity to feathers. Results showed that the first 50 amino acids of N terminal had a great influence on the expression and purification of protein Dgker. The crude enzyme solution of recombinant strain completely decomposed feathers in three days. The transparent circle on milk powder plate appeared more notable in crude enzyme solution of recombinant strain than that of empty strain. Dgker adapted to a wide range of temperatures and pH,among which the optimal temperature was 60℃ and the optimal pH was 5.0. Dgker can degrade feathers and thus will have great application space in the future industrial production and treatment of waste feathers.
中文关键词	角蛋白 ; 戈壁异常球菌 ; 角蛋白酶 ; 酶活
英文关键词	keratin Deinococcus gobiensis I-0 keratinase enzyme activity
语种	中文
国家	中国
收录类别	CSCD
WOS类目	MICROBIOLOGY
WOS研究方向	Microbiology
CSCD记录号	CSCD:6469317
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/239496
作者单位	1.西南科技大学生命科学与工程学院;;中国农业科学院生物技术研究所, ;;, 绵阳;;, ;;北京 621000;;100081; 2.中国农业科学院生物技术研究所, 北京 100081, 中国
推荐引用方式 GB/T 7714	耿秀秀,周正富,刘盈盈,等. 戈壁异常球菌角蛋白酶基因的克隆及功能研究[J],2019,35(3):65-70.
APA	耿秀秀,周正富,刘盈盈,平淑珍,&王劲.(2019).戈壁异常球菌角蛋白酶基因的克隆及功能研究.生物技术通报,35(3),65-70.
MLA	耿秀秀,et al."戈壁异常球菌角蛋白酶基因的克隆及功能研究".生物技术通报 35.3(2019):65-70.