Immobilization of watermelon (<Emphasis Type="Italic">Citrullus vulgaris</Emphasis>) urease in agarose gel for urea estimation |
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Authors: | Om Prakash Srilakshmi Puliga Lata Sheo Bachan Upadhyay |
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Institution: | (1) Department of Biochemistry, Faculty of Science, Banaras Hindu University, 221005 Varanasi, India |
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Abstract: | Urease from dehusked seeds of watermelon was immobilized in 1.5% agarose gel with 53.9% entrapment. There was negligible leaching
(<10% at 4°C) and the same gel membrane could repeatedly be used for seven days. The immobilization exhibited no apparent
change in the optimum pH but there was a significant decrease in the optimum temperature (50°C as compared to 65°C for soluble
urease). The immobilized urease revealed an apparentK
m of 9.3±0.3 mM; 1.2 times lower than the soluble enzyme (11.4±0.2 mM). Unlike soluble enzyme which was inhibited at 200 mM
urea, the immobilized urease was inhibited at 600 mM of urea and above, and about 47% activity was retained at 2 M urea. The
time-dependent thermal inactivation kinetics at 48 and 52°C was found to be biphasic, in which half of the initial activity
was destroyed more rapidly than the remaining half. These gel membranes were also used for estimating the urea content of
the blood samples from the University hospital. The results obtained matched well with those obtained by the usual method
employed in the clinical pathology laboratory. The significance of these observations is discussed. |
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Keywords: | urease watemelon Citrullus vulgaris immobilization agarose urease immobilization |
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