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Enhancement of nuclease P1 activity in low concentration of denaturants

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Relation http://ir.cftri.com/1505/
EMT-07-08
 
Title Enhancement of nuclease P1 activity in low concentration of denaturants
 
Creator Gangadhara, B.N.
Parigi Ramesh, Kumar
Prakash, V.
 
Subject 05 Enzymes
 
Description Nuclease P1 is a zinc-dependent endonuclease from the mold Penicillium citrinum. Nuclease P1 exhibits
an increase in activity in the presence of low concentrations of urea or GuHCI. At 0.05 M GuHCI or 1 M urea
the enzyme activity enhanced maximally by 3- and 3.9-fold. The kinetic parameters indicate a decrease in
Km with an increase in catalytic constant. The Km values for control and in presence of 0.05 M GuHCI or 1 M
urea are 1.11 mg, 0.86 mg and 0.60 mg, respectively. In presence of metal ions such as Cu2+ and Co2+, urea
or GuHCI treated enzyme still maintains the activity enhancement up to different extent. The far UV-CD
results point to a minor conformational change and fluorescence spectra reveal increase in the relative
fluorescence intensity at lower concentration. Thermal denaturation studies reveal increase in apparent
Tm from 75 C for control to 80 C and 77 C in presence of 0.1 M GuHCI or 0.5 M urea, respectively. The
above results show that the activation of nuclease PI in presence of low concentrations of denaturants is
mainly because of the increase in the catalytic constant suggesting that activation is due to a more open
and flexible conformation of the activated enzyme with increased catalytic efficiency.
 
Date 2008
 
Type Article
PeerReviewed
 
Format application/pdf
 
Language en
 
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Identifier http://ir.cftri.com/1505/2/Enzyme_and_Microbial_Technology_43_%282008%29_336-342.pdf
Gangadhara, B.N. and Parigi Ramesh, Kumar and Prakash, V. (2008) Enhancement of nuclease P1 activity in low concentration of denaturants. Enzyme and Microbial Technology, 43. pp. 336-342.