title: Gene cloning, expression, and characterization of a cyclic nucleotide phosphodiesterase from Arthrobacter sp. CGMCC 3584.
abstract: Based on thermal asymmetric interlaced polymerase chain reaction, the arpde gene encoding a cyclic nucleotide-specific phosphodiesterase was cloned from Arthrobacter sp. CGMCC 3584 for the first time. The 930-bp region encoded a 309-amino-acid protein with a molecular weight of 33.6 kDa. The recombinant ArPDE was able to hydrolyze 3',5'-cAMP, 3',5'-cGMP, and 2',3'-cAMP. The K m values of ArPDE for 3',5'-cAMP and 3',5'-cGMP were 6.82 and 12.82 mM, respectively. ArPDE was thermostable and displayed optimal activity at 45 °C and pH 7.5. The enzyme did not require any metal cofactors, although its activity was stimulated by 2 mM Co(2+) and inhibited by Zn(2+). Nucleotides, reducing agents, and sulfhydryl reagents had different inhibitory effects on the activity of ArPDE. NaF, the actual compound used to improve the industrial yield of cAMP, exhibited 62 % inhibitions at concentrations of 10 mM.
authors: College of Life Science and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing, 210009, People's Republic of China.
authors: Zhifang
authors: Z
authors: Zheng
authors: Mengzhu
authors: M
authors: Zhu
authors: Ying
authors: Y
authors: He
authors: Nan
authors: N
authors: Li
authors: Ting
authors: T
authors: Guo
authors: Yong
authors: Y
authors: Chen
authors: Jinglan
authors: J
authors: Wu
authors: Hanjie
authors: H
authors: Ying
authors: Jingjing
authors: J
authors: Xie
mesh_terms: D001173:Arthrobacter / Q000201:enzymology*; D003001:Cloning, Molecular; D006152:Cyclic GMP / Q000378:metabolism; D009712:Nucleotides, Cyclic / Q000378:metabolism*; D010727:Phosphoric Diester Hydrolases / Q000235:genetics / Q000378:metabolism*; D019163:Reducing Agents / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D009712:Nucleotides, Cyclic; D019163:Reducing Agents; D010727:Phosphoric Diester Hydrolases; D006152:Cyclic GMP