Expression, immobilization and enzymatic properties of glutamate decarboxylase fused to a cellulose-binding domain |
| |
Authors: | Park Hyemin Ahn Jungoh Lee Juwhan Lee Hyeokwon Kim Chunsuk Jung Joon-Ki Lee Hongweon Lee Eun Gyo |
| |
Affiliation: | Biotechnology Process Engineering Center, KRIBB, Daejeon 305-600, Korea; E-Mails: gohworld@kribb.re.kr (H.P.); ahnjo@kribb.re.kr (J.A.); jhlee@kribb.re.kr (J.L.); tntn7616@kribb.re.kr (H.L.); chskim@kribb.re.kr (C.K.); jkjung@kribb.re.kr (J.-K.J.); hwlee@kribb.re.kr (H.L.). |
| |
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 S(3)N(10) 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 nmol(CBD-GAD)/g(Avicel) 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 |
本文献已被 PubMed 等数据库收录! |
|