机构：[1]Department of Central Laboratory, Shenzhen Hospital, Beijing University of Chinese Medicine, Shenzhen, 518100, Guangdong, China北京大学深圳医院深圳市康宁医院深圳医学信息中心[2]Institute of Translational Medicine, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, 518035, China深圳市第二人民医院深圳医学信息中心
5'- nucleotidase (5'-NT) is a key enzyme in nucleoside/nucleotide metabolic pathway, it plays an important role in the biosynthesis of cordycepin in caterpillar fungus. In this study, a 5'-NT gene was identified and mined from genomic DNA of caterpillar fungus, which was 1968 bp in length and encoded 656 amino acid residues. The recombinant 5'-NT was first time heterologously expressed in Pichia pastoris GS115, subsequently purified and functionally characterized. The optimal reaction temperature for 5'-NT was 35 degrees C, and it retained 52.8% of its residual activity after incubation at 50 degrees C for 1 h. The optimal reaction pH was 6.0 and it exhibited high activity over a neutral pH range. Furthermore, 5'-NT exhibited excellent K-m (1.107 mM), V-max (0.113 mu mol/mg(.)min) and k(cat) (4.521 S-1) values compared with other typical 5'-nucleotidase. Moreover, substrate specificity analyses indicated that 5'-NT exhibited different phosphatase activity towards the substrates containing different basic groups. The work presented here could be useful to 5'-NT applications and provide more scientific basis and new ideas for the biosynthesis of artificial control cordycepin.
基金：
National Natural Science Foundation of ChinaNational Natural Science Foundation of China [81803652]; Natural Science Foundation of Guangdong ProvinceNational Natural Science Foundation of Guangdong Province [2018A0303100007]; Shenzhen Foundation of Health and Family Planning Commission [SZBC2018016]; China Postdoctoral Science FoundationChina Postdoctoral Science Foundation [2018M633261]
语种：
外文
被引次数：
WOS：
PubmedID：
中科院(CAS)分区：
出版当年[2019]版：
大类|4 区生物
小类|4 区生化研究方法4 区生化与分子生物学4 区生物工程与应用微生物
最新[2025]版：
大类|4 区生物学
小类|4 区生化研究方法4 区生化与分子生物学4 区生物工程与应用微生物
JCR分区：
出版当年[2018]版：
Q4BIOCHEMICAL RESEARCH METHODSQ4BIOCHEMISTRY & MOLECULAR BIOLOGYQ4BIOTECHNOLOGY & APPLIED MICROBIOLOGY
最新[2023]版：
Q4BIOCHEMICAL RESEARCH METHODSQ4BIOCHEMISTRY & MOLECULAR BIOLOGYQ4BIOTECHNOLOGY & APPLIED MICROBIOLOGY
第一作者机构：[1]Department of Central Laboratory, Shenzhen Hospital, Beijing University of Chinese Medicine, Shenzhen, 518100, Guangdong, China[2]Institute of Translational Medicine, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, 518035, China[*1]Department of Central Laboratory, Shenzhen Hospital, Beijing University of Chinese Medicine, Shenzhen, 518100, Guangdong, China.
通讯作者：
通讯机构：[1]Department of Central Laboratory, Shenzhen Hospital, Beijing University of Chinese Medicine, Shenzhen, 518100, Guangdong, China[2]Institute of Translational Medicine, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, 518035, China[*1]Department of Central Laboratory, Shenzhen Hospital, Beijing University of Chinese Medicine, Shenzhen, 518100, Guangdong, China.
推荐引用方式(GB/T 7714)：
Lin Shan,Zhou Cuibing,Zhang Hancheng,et al.Expression, purification and characterization of 5 '-nucleotidase from caterpillar fungus by efficient genome-mining[J].PROTEIN EXPRESSION AND PURIFICATION.2020,168:doi:10.1016/j.pep.2019.105566.
APA：
Lin, Shan,Zhou, Cuibing,Zhang, Hancheng&Cai, Zhiming.(2020).Expression, purification and characterization of 5 '-nucleotidase from caterpillar fungus by efficient genome-mining.PROTEIN EXPRESSION AND PURIFICATION,168,
MLA：
Lin, Shan,et al."Expression, purification and characterization of 5 '-nucleotidase from caterpillar fungus by efficient genome-mining".PROTEIN EXPRESSION AND PURIFICATION 168.(2020)
