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A Study on the Comparative Stability of Insoluble Complexes of Glucose Oxidase Obtained with Concanavalin A and Specific Polyclonal Antibodies

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Summary

The formation of insoluble complexes of glycoenzymes with lectins and antibodies is one of the simplest methods of enzyme immobilization. Insoluble complexes of glucose oxidase were simply obtained by mixing the enzyme with concanavalin A or a specific polyclonal antibodies solution. The concanavalin A and immunocomplexes of glucose oxidase retained more than 80% of the original enzyme activity. Expression of very high enzyme activity in insoluble complexes suggested that these aggregates were quite porous and easily accessible to substrates. Insoluble complexes of glucose oxidase showed very high stability against denaturation induced by pH, temperature, urea and water-miscible organic solvents. Complexes of glucose oxidase obtained with concanavalin A and glycosyl-specific antiglucose oxidase polyclonal antibodies were quite comparable in stability while complexes prepared using polyclonal antibodies raised against the native glucose oxidase were slightly less stable. The complexes of glucose oxidase obtained with glycosyl-specific antiglucose oxidase polyclonal antibodies showed very high stability against inactivation mediated by exposure to water-miscible organic solvents. Insoluble complexes of glucose oxidase were cross-linked with glutaraldehyde to maintain their integrity in the presence of substrates. The cross-linking of complexes resulted in a slight decrease in enzyme activity but showed a pronounced enhancement in stability against various forms of denaturation.

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Abbreviations

Con A:

concanavalin A

GOD:

glucose oxidase

IDA:

iminodiacetate

IgG:

immunoglobulin G

THF:

tetrahydrofuran

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Authors and Affiliations

  1. Department of Biochemistry, Shere Kashmir Institute of Medical Sciences (SKIMS) Soura, Srinagar, Jammu and Kashmir, India

    Ulfat Jan

  2. Department of Biochemistry, Faculty of Life Science, Aligarh Muslim University, 202002, Aligarh, India

    Amjad Ali Khan & Qayyum Husain

Authors
  1. Ulfat Jan
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  2. Amjad Ali Khan
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  3. Qayyum Husain
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Correspondence to Qayyum Husain.

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Jan, U., Khan, A.A. & Husain, Q. A Study on the Comparative Stability of Insoluble Complexes of Glucose Oxidase Obtained with Concanavalin A and Specific Polyclonal Antibodies. World J Microbiol Biotechnol 22, 1033–1039 (2006). https://doi.org/10.1007/s11274-005-3208-6

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  • DOI: https://doi.org/10.1007/s11274-005-3208-6

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