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Determination of serum lactate with alkylamine glass bound lactate oxidase

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Title Determination of serum lactate with alkylamine glass bound lactate oxidase
 
Creator Suman
Pundir, C S
 
Subject Lactate
Lactate oxidase
Immobilization
Alkylamine glass
Serum
Lactic acid determination
 
Description 186-189
Commercial lactate oxidase (Lactate:O₂; oxidoreductase EC 1.1.3.2) from Pediococcus species was immobilized on to al-kylamine glass beads (pore diameter 55 nm) through glutaralde-hyde coupling with a conjugation yield of 3.2 mg/g support and 105% retention of initial activity. Immobilized enzyme showed maximum activity at pH 6.5, when incubated at 40ºC for 12 min and was used for determination of lactic acid in serum. The H₂O₂ generated from serum lactate by immobilized enzyme was meas-ured colorimetrically at 565 nm by its oxidative coupling with
4-aminoantipyrine and N,N'-dimethyaniline catalyzed by horse-radish peroxidase. A linear relationship was observed between A₅₆₅ and lactic acid concentration ranging from 0.075 mM to
10 mM. The minimum detection limit of the method was
0.075 mM, which was better than that of enzymic colorimetric method employing free enzyme (0.2 mM). Within day and
between day coefficient of variations were
 
Date 2009-03-30T07:43:16Z
2009-03-30T07:43:16Z
2005-06
 
Type Article
 
Identifier 0301-1208
http://hdl.handle.net/123456789/3520
 
Language en_US
 
Relation C 12 N 11/00, C 12 Q
 
Publisher CSIR
 
Source IJBB Vol.42(3) [June 2005]