Polyphosphates and exopolyphosphatase activities in the yeast Saccharomyces cerevisiae under overexpression of homologous and heterologous PPN1 genes

被引:20
作者
Eldarov, M. A. [1 ]
Baranov, M. V. [1 ]
Dumina, M. V. [1 ]
Shgun, A. A. [1 ]
Andreeva, N. A. [2 ]
Trilisenko, L. V. [2 ]
Kulakovskaya, T. V. [2 ]
Ryasanova, L. P. [2 ]
Kulaev, I. S. [2 ]
机构
[1] Russian Acad Sci, Ctr Bioengn, Moscow 117312, Russia
[2] Russian Acad Sci, Skryabin Inst Biochem & Physiol Microorganisms, Pushchino 142290, Moscow Region, Russia
基金
俄罗斯基础研究基金会;
关键词
polyphosphates; exopolyphosphatase; PPN1; gene; Saccharomyces cerevisiae; Acremonium chrysogenum; transformation; MOLECULAR-MASS EXOPOLYPHOSPHATASE; INORGANIC POLYPHOSPHATE; ENDOPOLYPHOSPHATASE GENE; INACTIVATION; PURIFICATION; CYTOSOL; PPX1;
D O I
10.1134/S0006297913080129
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The role of exopolyphosphatase PPN1 in polyphosphate metabolism in fungi has been studied in strains of Saccharomyces cerevisiae transformed by the yeast PPN1 gene and its ortholog of the fungus Acremonium chrysogenum producing cephalosporin C. The PPN1 genes were expressed under a strong constitutive promoter of the gene of glycerol aldehyde-triphosphate dehydrogenase of S. cerevisiae in the vector pMB1. The yeast strain with inactivated PPN1 gene was transformed by the above vectors containing the PPN1 genes of S. cerevisiae and A. chrysogenum. Exopolyphosphatase activity in the transformant with the yeast PPN1 increased 28- and 11-fold compared to the mutant and parent PPN1 strains. The amount of polyphosphate in this transformant decreased threefold. Neither the increase in exopolyphosphatase activity nor the decrease in polyphosphate content was observed in the transformant with the orthologous PPN1 gene of A. chrysogenum, suggesting the absence of the active form of PPN1 in this transformant.
引用
收藏
页码:946 / 953
页数:8
相关论文
共 26 条
[1]   High molecular mass exopolyphosphatase from the cytosol of the yeast Saccharomyces cerevisiae is encoded by the PPN1 gene [J].
Andreeva, N. A. ;
Kulakovskaya, T. V. ;
Kulaev, I. S. .
BIOCHEMISTRY-MOSCOW, 2006, 71 (09) :975-977
[2]   Purification and properties of exopolyphosphatase from the cytosol of Saccharomyces cerevisiae not encoded by the PPX1 gene [J].
Andreeva, NA ;
Kulakovskaya, TV ;
Kulaev, IS .
BIOCHEMISTRY-MOSCOW, 2004, 69 (04) :387-393
[3]   PURIFICATION AND CHARACTERIZATION OF HIGHLY-ACTIVE AND STABLE POLYPHOSPHATASE FROM SACCHAROMYCES-CEREVISIAE CELL-ENVELOPE [J].
ANDREEVA, NA ;
OKOROKOV, LA .
YEAST, 1993, 9 (02) :127-139
[4]  
[Anonymous], 2004, BIOCH INORGANIC POLY
[5]   ASSAY OF PROTEINS IN PRESENCE OF INTERFERING MATERIALS [J].
BENSADOUN, A ;
WEINSTEIN, D .
ANALYTICAL BIOCHEMISTRY, 1976, 70 (01) :241-250
[6]  
CAEN J, 2010, THROMBOSIS HEMOSTASI, V8, P1
[7]   Evolution of acidocalcisomes and their role in polyphosphate storage and osmoregulation in eukaryotic microbes [J].
Docampo, Roberto ;
Ulrich, Paul ;
Moreno, Silvia N. J. .
PHILOSOPHICAL TRANSACTIONS OF THE ROYAL SOCIETY B-BIOLOGICAL SCIENCES, 2010, 365 (1541) :775-784
[8]   Alkaline sphingomyelinase: An old enzyme with novel implications [J].
Duan, Rui-Dong .
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR AND CELL BIOLOGY OF LIPIDS, 2006, 1761 (03) :281-291
[9]   Optimization of ordered plasmid assembly by gap repair in Saccharomyces cerevisiae [J].
Eckert-Boulet, Nadine ;
Pedersen, Mette Louise ;
Krogh, Berit Olsen ;
Lisby, Michael .
YEAST, 2012, 29 (08) :323-334
[10]  
Eldarov M. A., 2000, Patent of the Russian Federation, Patent No. 2150501