摘要
克隆得到梅花鹿过氧化氢酶基因序列,已提交Genbank登录(HQ877674).基因编码区全长1 584 bp,编码527个氨基酸,理论计算分子量为60 027.4 Da,理论计算等电点为6.67,预测蛋白质结构中不含有二硫键,在蛋白质序列中,具有过氧化氢酶家族活性中心保守序列和过氧化氢酶家族亚铁血红素保守序列,其DNA序列和蛋白质序列均与Bos taurus来源过氧化氢酶的同源关系最为接近.使用pPICZαC质粒和毕赤酵母GS115菌株,成功异源表达该基因,对诱导表达的发酵上清液进行活性测定,酶活为463 U·mL-1.
We have obtained the full length gene sequence of Cervus nippon catalase successfully, which has been submitted to Genbank under the accession number HQ877674. The length of this gene is 1 584 bp, which encodes 527 amino acids, the theoretical MW is 60 027.4 Da and the theoretical pI is 6.67, there is no disulfide bond in this protein as the prediction, catalase proximal active site signture and catalase proximal heme -ligand signture were found, and, both the DNA sequence and protein sequence of this work are homologous to the catalase from Bos taurus. Heterologous protein expression was carried out with pPICZαC plasmid and Pichia pastoris GS115, the liquid supernatant of the induced recombinant contains 463 U ·mL- 1 catalase activity.
出处
《福州大学学报（自然科学版）》
CAS
CSCD
北大核心
2014年第1期154-160,共7页
Journal of Fuzhou University(Natural Science Edition)
基金
福建省自然科学基金资助项目(2011J01218)
福建省高校产学合作科技重大项目(2013N5005)
福州大学科技发展基金资助项目(2013-XY-17)
关键词
梅花鹿
过氧化氢酶
克隆
表达
Cervus nippon
catalase
cloning
expression
