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Purification and properties of polyphosphatase from Saccharomyces cerevisiae cytosol
Author(s) -
Andreeva Nadezhda,
Kulakovskaya Tatjana,
Sidorov Igor,
Karpov Alexander,
Kulaev Igor
Publication year - 1998
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/(sici)1097-0061(19980315)14:4<383::aid-yea232>3.0.co;2-s
Subject(s) - divalent , cytosol , polyphosphate , saccharomyces cerevisiae , enzyme , biology , biochemistry , hydrolysis , molecular mass , yield (engineering) , yeast , phosphate , chemistry , organic chemistry , materials science , metallurgy
A homogenous polyphosphatase preparation was obtained from Saccharomyces cerevisiae cytosol with a 3·8% yield and 3540‐fold purification. The enzyme hydrolysed polyphosphate (polyP) with various chain lengths, including polyP 3 , and split P i off the end of the chain. It was inactive with respect to ATP, PP i , and p ‐nitrophenylphosphate. Its specific activity with polyP 15 was 283 U/mg protein. The polyphosphatase was a monomeric protein with a molecular mass of 40 kDa. This enzyme was inactive without divalent cations and with Cu 2+ and Ca 2+ . The ability of other divalent cations to activate the enzyme decreased in the following order: Co 2+ >Mn 2+ >Mg 2+ >Zn 2+ . A kinetic model of the hydrolysis of polyP 3 and action of Mg 2+ is proposed. © 1998 John Wiley & Sons, Ltd.