Gamma-aminobutyric Acid production using immobilized glutamate decarboxylase followed by downstream processing with cation exchange chromatography

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Title
Gamma-aminobutyric Acid production using immobilized glutamate decarboxylase followed by downstream processing with cation exchange chromatography
Author(s)
Seungwoon Lee; Jungoh AhnYeon Gu Kim; Joon Ki Jung; Hong-Weon LeeEun Gyo Lee
Bibliographic Citation
International Journal of Molecular Sciences, vol. 14, no. 1, pp. 1728-1739
Publication Year
2013
Abstract
We have developed a gamma-aminobutyric acid (GABA) production technique using his-tag mediated immobilization of Escherichia coli-derived glutamate decarboxylase (GAD), an enzyme that catalyzes the conversion of glutamate to GABA. The GAD was obtained at 1.43 g/L from GAD-overexpressed E. coli fermentation and consisted of 59.7% monomer, 29.2% dimer and 2.3% tetramer with a 97.6% soluble form of the total GAD. The harvested GAD was immobilized to metal affinity gel with an immobilization yield of 92%. Based on an investigation of specific enzyme activity and reaction characteristics, glutamic acid (GA) was chosen over monosodium glutamate (MSG) as a substrate for immobilized GAD, resulting in conversion of 2.17 M GABA in a 1 L reactor within 100 min. The immobilized enzymes retained 58.1% of their initial activities after ten consecutive uses. By using cation exchange chromatography followed by enzymatic conversion, GABA was separated from the residual substrate and leached GAD. As a consequence, the glutamic acid was mostly removed with no detectable GAD, while 91.2% of GABA was yielded in the purification step.
Keyword
Cation exchange chromatographyGamma aminobutyric acidGlutamate decarboxylaseGlutamic acidImmobilization
ISSN
1422-0067
Publisher
MDPI
DOI
http://dx.doi.org/10.3390/ijms14011728
Type
Article
Appears in Collections:
Division of Bio Technology Innovation > BioProcess Engineering Center > 1. Journal Articles
Division of Biomedical Research > Biotherapeutics Translational Research Center > 1. Journal Articles
Division of Bio Technology Innovation > 1. Journal Articles
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