Bacillus subtilis 5′-nucleotidases with various functions and substrate specificities

BMC Microbiology

Research Article

Ayane Natsume¹  Shogo Nishihata¹  Junko Kuse¹  Ayako Terakawa¹  Atsushi Okada¹  Shinji Takenaka²  Shu Ishikawa³  Kosei Tanaka⁴  Ken-ichi Yoshida⁵ 

[1] Department of Agrobioscience, Kobe University, 1-1 Rokkodai, Nada, 657-8501, Kobe, Hyogo, Japan;Department of Agrobioscience, Kobe University, 1-1 Rokkodai, Nada, 657-8501, Kobe, Hyogo, Japan;Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, 657-8501, Kobe, Hyogo, Japan;Department of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, 657-8501, Kobe, Hyogo, Japan;Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, 657-8501, Kobe, Hyogo, Japan;Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, 657-8501, Kobe, Hyogo, Japan;Department of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, 657-8501, Kobe, Hyogo, Japan;

关键词: Bacillus subtilis;    Haloacid dehalogenase superfamily;    Inositol monophosphatase;    Inositol phosphate;    Nucleoside/nucleotide metabolism;    5′-nucleotidase;    Oxidative stress;    Phosphatase;    Protein motif;   

DOI  :  10.1186/s12866-016-0866-5

 received in 2016-09-08, accepted in 2016-10-15,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundIn Escherichia coli, nagD, yrfG, yjjG, yieH, yigL, surE, and yfbR encode 5′-nucleotidases that hydrolyze the phosphate group of 5′-nucleotides. In Bacillus subtilis, genes encoding 5′-nucleotidase have remained to be identified.ResultsWe found that B. subtilis ycsE, araL, yutF, ysaA, and yqeG show suggestive similarities to nagD. Here, we expressed them in E. coli to purify the respective His6-tagged proteins. YcsE exhibited significant 5′-nucleotidase activity with a broader specificity, whereas the other four enzymes had rather weak but suggestive activities with various capacities and substrate specificities. In contrast, B. subtilis yktC shares high similarity with E. coli suhB encoding an inositol monophosphatase. YktC exhibited inositol monophosphatase activity as well as 5′-nucleotidase activity preferential for GMP and IMP. The ycsE, yktC, and yqeG genes are induced by oxidative stress and were dispensable, although yqeG was required to maintain normal growth on solid medium. In the presence of diamide, only mutants lacking yktC exhibited enhanced growth defects, whereas the other mutants without ycsE or yqeG did not.ConclusionsAccordingly, in B. subtilis, at least YcsE and YktC acted as major 5′-nucleotidases and the four minor enzymes might function when the intracellular concentrations of substrates are sufficiently high. In addition, YktC is involved in resistance to oxidative stress caused by diamide, while YqeG is necessary for normal colony formation on solid medium.

【 授权许可】

CC BY   
© The Author(s). 2016