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Purification and characterization of 6-phosphogluconate dehydrogenase (6-PGD) from grass carp (<i style="mso-bidi-font-style: normal">Ctenopharyngodon idella</i>) hepatopancreas

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Title Purification and characterization of 6-phosphogluconate dehydrogenase (6-PGD) from grass carp (Ctenopharyngodon idella) hepatopancreas
 
Creator Sun, Lin-Dan
Luo, Zhi
Hu, Wei
Zhuo, Mei-Qin
Zheng, Jia-Lang
Chen, Qi-Liang
Liang, Xu-Fang
Xiong, Bang-Xi
 
Subject Ctenopharyngodon idella
Grass carp
6-Phosphogluconate dehydrogenase
Purification
Kinetic behavior
Metal ions inhibition
 
Description 554-561
6-Phosphogluconate dehydrogenase (6-PGD, E.C.: 1.1.1.44) was
purified and characterized from the hepatopancreas of grass carp (Ctenopharyngodon idella)
for the first time. Grass carp represents the second largest aquaculture industry
in the world after silver carp, constituting 14.7% of the world aquaculture
production, with an average annual increase of 14% in China, mainly as a source of food.
The purification procedure involved a single 2’, 5’-ADP-Sepharose
4B affinity chromatographic step by using different elution buffers. The enzyme
was purified 309-fold with a specific activity of
5.259 U/mg protein and yield of 68%. The purity and subunit molecular weights
of the 6-PGD were checked on SDS-PAGE and purified enzyme showed a single band
on the gel. The subunit molecular mass was 57 kDa, with an optimum pH,
temperature and ionic strength at 7.96, 50oC and 100 mM Tris-HCl, respectively. The Km values of 6-PGA and NADP+ were 0.019 and 0.0052 mM, respectively, while Vm of 6-PGA and NADP+
was 0.69 U/ml. Dissociation constants (Ki)
for 6-PGA and NADP+ were 2.05 and 0.12 mM, respectively. NADPH inhibited the enzyme in a
competitive manner and its Ki
value was 0.032 mM. The
Cu2+, Zn2+, Cd2+ and Al3+ showed
inhibitory effects on the enzyme with IC50 values of 0.293, 0.099,
0.045 and 1.526 mM,
respectively. All tested metals inhibited the enzyme in a competitive manner,
indicating that these metals might be toxic even at low concentrations for the
6-PGD. As the fish is one of valuable foodstuff of animal sources for human
consumption, under certain environmental conditions, metal ions accumulated in
fish up to a lethal concentration may be harmful for human health. Therefore,
it is impending to reduce the concentration of metal ions in contaminated lakes
and rivers for fishery and also for human health.
 
Date 2013-12-27T11:54:52Z
2013-12-27T11:54:52Z
2013-12
 
Type Article
 
Identifier 0975-0959 (Online); 0301-1208 (Print)
http://hdl.handle.net/123456789/25174
 
Language en_US
 
Rights CC Attribution-Noncommercial-No Derivative Works 2.5 India
 
Publisher NISCAIR-CSIR, India
 
Source IJBB Vol.50(6) [December 2013]