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<span style="font-size:11.0pt;mso-bidi-font-size: 10.0pt;font-family:"Times New Roman";mso-fareast-font-family:"Times New Roman"; mso-ansi-language:EN-GB;mso-fareast-language:EN-US;mso-bidi-language:AR-SA" lang="EN-GB">Kinetic behaviour of calf intestinal alkaline phosphatase with <i style="mso-bidi-font-style: normal">p</i>NPP</span>

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Title Kinetic behaviour of calf intestinal alkaline phosphatase with pNPP
 
Creator Chaudhuri, Gouri
Chatterjee, Saswata
Venu-Babu, P
Ramasamy, K
Thilagaraj, W Richard
 
Subject Calf intestinal alkaline phosphatase
Glycine-NaOH buffer
Hydrolysis
p-Nitrophenyl phosphate
Temperature
Tris-HCl buffer
Bioremediation
 
Description 64-71
The hydrolysis of p-nitrophenyl
phosphate (pNPP) by calf intestinal
alkaline phosphatase (CIAP) was investigated
with respect to kinetic parameters such as Vmax,
Km and Kcat under varying pH,
buffers, substrate concentration, temperature and period of incubation. Highest
activity was obtained with Tris-HCl at pH 11, while in the case of glycine-NaOH
buffer the peak activity was recorded at pH 9.5. The enzyme showed the
following kinetic characteristics with pNPP in 50 mM Tris-HCl at pH 11
and 100 mM glycine-NaOH at pH 9.5 at an incubation temperature of 37°C: Vmax, 3.12
and 1.6 µmoles
min-1 unit-1; Km,
7.6 × 10-4 M and 4 × 10-4 M; and Kcat, 82.98 s-1 and 42.55
s-1, respectively. CIAP displayed a high
temperature optimum of 45°C at pH 11. The kinetic behaviour of the enzyme under
different parameters suggested that
the enzyme might undergo subtle conformational changes in response to the
buffers displaying unique characteristics. Bioprecipitation of Cu2+
from 50 ppm of CuCl2 solution was studied where 64.3% of precipitation
was obtained.
Pi generated from CIAP-mediated hydrolysis of pNPP was found to bind with copper and precipitated as
copper-phosphate. Thus, CIAP could be used as a test candidate in
bioremediation of heavy metals from industrial wastes through generation of
metal-phosphate complexes.


 
Date 2013-02-23T11:26:08Z
2013-02-23T11:26:08Z
2013-02
 
Type Article
 
Identifier 0975-0959 (Online); 0301-1208 (Print)
http://hdl.handle.net/123456789/16064
 
Language en_US
 
Rights CC Attribution-Noncommercial-No Derivative Works 2.5 India
 
Publisher NISCAIR-CSIR, India
 
Source IJBB Vol.50(1) [February 2013]