Int. J. Mol. Sci. 2012, 13(1), 358-368; doi:10.3390/ijms13010358
Article

Expression, Immobilization and Enzymatic Properties of Glutamate Decarboxylase Fused to a Cellulose-Binding Domain

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Received: 14 November 2011; in revised form: 15 December 2011 / Accepted: 19 December 2011 / Published: 28 December 2011
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract: Escherichia coli-derived glutamate decarboxylase (GAD), an enzyme that catalyzes the conversion of glutamic acid to gamma-aminobutyric acid (GABA), was fused to the cellulose-binding domain (CBD) and a linker of Trichoderma harzianum endoglucanase II. To prevent proteolysis of the fusion protein, the native linker was replaced with a S3N10 peptide known to be completely resistant to E. coli endopeptidase. The CBD-GAD expressed in E. coli was successfully immobilized on Avicel, a crystalline cellulose, with binding capacity of 33 ± 2 nmolCBD-GAD/gAvicel and the immobilized enzymes retained 60% of their initial activities after 10 uses. The results of this report provide a feasible alternative to produce GABA using immobilized GAD through fusion to CBD.
Keywords: GAD; cellulose-binding domain; fusion protein; immobilization
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MDPI and ACS Style

Park, H.; Ahn, J.; Lee, J.; Lee, H.; Kim, C.; Jung, J.-K.; Lee, H.; Lee, E.G. Expression, Immobilization and Enzymatic Properties of Glutamate Decarboxylase Fused to a Cellulose-Binding Domain. Int. J. Mol. Sci. 2012, 13, 358-368.

AMA Style

Park H, Ahn J, Lee J, Lee H, Kim C, Jung J-K, Lee H, Lee EG. Expression, Immobilization and Enzymatic Properties of Glutamate Decarboxylase Fused to a Cellulose-Binding Domain. International Journal of Molecular Sciences. 2012; 13(1):358-368.

Chicago/Turabian Style

Park, Hyemin; Ahn, Jungoh; Lee, Juwhan; Lee, Hyeokwon; Kim, Chunsuk; Jung, Joon-Ki; Lee, Hongweon; Lee, Eun Gyo. 2012. "Expression, Immobilization and Enzymatic Properties of Glutamate Decarboxylase Fused to a Cellulose-Binding Domain." Int. J. Mol. Sci. 13, no. 1: 358-368.

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