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Immobilization of Urease from Pigeonpea (Cajanus cajan) on Agar Tablets and Its Application in Urea Assay

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Abstract

The pigeonpea urease was immobilized on agar, a common gelling substance. The tablet strips were used as moulds to cast agar tablets of uniform shape and size. The time and temperature of solidification of agar was 6 min and 44 °C, respectively. The 5 % agar (w/v) and 0.019 mg protein/agar tablet yielded an optimum immobilization of 51.7%. The optimum pH was shifted through 0.2 U (from 7.3 to 7.5) towards basic side upon immobilization. The optimum temperature of soluble and immobilized urease was 30 °C and 60 °C, respectively, showing the improvement in thermal stability of urease. There was an increase in K m from 3.23 to 5.07 mM after immobilization. The half-lives of soluble and immobilized urease were 21 and 53 days, respectively, at pH 7.3 and 4 °C. The urea was estimated in different blood samples with the help of immobilized urease and the results were consistent with those from clinical pathology laboratory through an autoanalyzer® (Zydus Co., Rome, Italy).

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Acknowledgements

Facilities provided by School of Biotechnology are gratefully acknowledged. S. K. and R.C.R.K. are thankful to Council for Scientific and Industrial Research and Indian Council of Medical Research, New Delhi, India for financial support in the form of Senior Research Fellowships.

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Authors and Affiliations

  1. School of Biotechnology, Faculty of Science, Banaras Hindu University, Varanasi, 221005, India

    Swati Mulagalapalli, Sandeep Kumar, Ravi Charan Reddy Kalathur & Arvind M. Kayastha

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  1. Swati Mulagalapalli
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  2. Sandeep Kumar
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  3. Ravi Charan Reddy Kalathur
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  4. Arvind M. Kayastha
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Correspondence to Arvind M. Kayastha.

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Mulagalapalli, S., Kumar, S., Kalathur, R.C.R. et al. Immobilization of Urease from Pigeonpea (Cajanus cajan) on Agar Tablets and Its Application in Urea Assay. Appl Biochem Biotechnol 142, 291–297 (2007). https://doi.org/10.1007/s12010-007-0022-7

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  • DOI: https://doi.org/10.1007/s12010-007-0022-7

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