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

Hyemin Park † email, Jungoh Ahn † email, Juwhan Leeemail, Hyeokwon Leeemail, Chunsuk Kimemail, Joon-Ki Jungemail, Hongweon Leeemail and Eun Gyo Lee* email
Biotechnology Process Engineering Center, KRIBB, Daejeon 305-600, Korea † These authors contributed equally to this work.
* Author to whom correspondence should be addressed.
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)
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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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Cite This Article

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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