Purification and properties of lactate dehydrogenase from liver of Uromastix hardwickii

Lactate dehydrogenase isoenzyme-1 was purified from liver of Uromastix hardwickii using colchicine-Sepharose and heat-inactivation methods. The crude enzyme showed four isoenzymes by agarose gel electrophoresis (AGE). The purified enzyme showed a single band after native AGE and SDS-PAGE correspondi...

Full description

Saved in:
Bibliographic Details
Published in:Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 1995-05, Vol.111 (1), p.27-34
Main Authors: Javed, Masood-Ul-Hassan, Yousuf, Farzana Abubakar, Hussain, Abida Nazir, Ishaq, Muhammad, Waqar, Mohammad Anwar
Format: Article
Language:English
Subjects:
Animals
Cations, Divalent
Electrophoresis, Agar Gel
Electrophoresis, Polyacrylamide Gel
Enzyme Stability
Glutamic Acid - pharmacology
Hot Temperature
Hydrogen-Ion Concentration
Isoenzymes
Kinetics
L-Lactate Dehydrogenase - chemistry
L-Lactate Dehydrogenase - isolation & purification
L-Lactate Dehydrogenase - metabolism
Lactates - metabolism
Lactic Acid
LDH isoenzyme-1
Liver
Liver - enzymology
Lizards
Molecular Weight
NAD - metabolism
Oxalates - pharmacology
Oxalic Acid
Properties
Purification
Pyruvates - metabolism
Pyruvic Acid
Uromastix hardwickii
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Lactate dehydrogenase isoenzyme-1 was purified from liver of Uromastix hardwickii using colchicine-Sepharose and heat-inactivation methods. The crude enzyme showed four isoenzymes by agarose gel electrophoresis (AGE). The purified enzyme showed a single band after native AGE and SDS-PAGE corresponding to a molecular weight of 34 kDa. The enzyme did not bind with DEAE-Sepharose at pH 7.2. The optimum pH for forward reaction was 7.5, while for reverse reaction, the maximum activity was at pH 9.5. The K m values for pyruvate, NADH, lactate and NAD + were 0.105, 0.045, 9.0 and 0.011 mM, respectively. The pyruvate showed maximum activity at about 150 μM and then starts showing inhibition at higher concentration. Pre-heating of enzyme showed that it was stable at 8°C for 30 min and at 100°C it became inactive immediately. Oxalate, glutamate, Cu 2+, Co 2+, Mn 2+, and Mg 2+ have shown inhibitory effects both for forward- and reverse-reactions. From these properties, we suggest that LDH-1 from Uromastix liver may be quite different from that of other vertebrates.
ISSN:1096-4959
1879-1107
DOI:10.1016/0305-0491(94)00230-R