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Immobilization of Escherichia coli novablue γ-glutamyltranspeptidase in Ca-alginate-k-carrageenan beads

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An Erratum to this article was published on 23 October 2008

Abstract

The recombinant Escherichia coli γ-glutamyltranspeptidase (EcGGT) was immobilized in Ca-alginate-k-carrageenan beads. Effects of alginate concentration, amount of loading enzyme, and bead size on the entrapped activity were investigated. Optimum alginate concentration for EcGGT immobilization was found to be 2% (w/v). Using a loading enzyme concentration of 1.5 mg/g alginate, maximum enzyme activity was observed. With increase in bead size from 1.9 to 3.1 mm, the immobilization efficiency was decreased significantly because of mass transfer resistance. Thermal stability of the free EcGGT was increased as a result of the immobilization. Ca-alginate-k-carrageenan-EcGGT beads were suitable for up to six repeated uses, losing only 45% of their initial activity. Upon 30 days of storage the preserved activity of free and immobilized enzyme were found as 4% and 68%, respectively. The synthesis of l-theanine was performed in 50 mM Tris–HCl buffer (pH 10) containing 25 mM l-glutamine, 40 mM ethylamine, and 1.5 mg EcGGT/g alginate at 40°C for 12 h, and a conversion rate of 27% was achieved.

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Acknowledgements

The financial support (NSC 95-2313-B-415-012-MY3) from National Science Council of Taiwan, Republic of China is thankfully acknowledged.

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Authors and Affiliations

  1. Department of Anesthesiology, Chang Gung Memorial Hospital, Kaohsiung, Taiwan

    Chih-Peng Hung

  2. Department of Food and Nutrition, Hungkuang University, Taichung, 433, Taiwan

    Huei-Fen Lo

  3. Institute of Molecular Biology, National Chung Hsing University, Taichung, 402-27, Taiwan

    Wen-Hwei Hsu

  4. Department of Applied Chemistry, National Chiayi University, 300 University Road, Chiayi, 60083, Taiwan

    Shih-Chun Chen & Long-Liu Lin

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  1. Chih-Peng Hung
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  2. Huei-Fen Lo
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Correspondence to Long-Liu Lin.

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An erratum to this article can be found at http://dx.doi.org/10.1007/s12010-008-8393-y

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Hung, CP., Lo, HF., Hsu, WH. et al. Immobilization of Escherichia coli novablue γ-glutamyltranspeptidase in Ca-alginate-k-carrageenan beads. Appl Biochem Biotechnol 150, 157–170 (2008). https://doi.org/10.1007/s12010-008-8244-x

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