title: Characterization and immobilization on nickel-chelated Sepharose of a glutamate decarboxylase A from Lactobacillus brevis BH2 and its application for production of GABA.
abstract: A gene encoding glutamate decarboxylase A (GadA) from Lactobacillus brevis BH2 was expressed in a His-tagged form in Escherichia coli cells, and recombinant protein exists as a homodimer consisting of identical subunits of 53 kDa. GadA was absolutely dependent on the ammonium sulfate concentration for catalytic activity and secondary structure formation. GadA was immobilized on the metal affinity resin with an immobilization yield of 95.8%. The pH optima of the immobilized enzyme were identical with those of the free enzyme. However, the optimum temperature for immobilized enzyme was 5 °C higher than that for the free enzyme. The immobilized GadA retained its relative activity of 41% after 30 reuses of reaction within 30 days and exhibited a half-life of 19 cycles within 19 days. A packed-bed bioreactor with immobilized GadA showed a maximum yield of 97.8% GABA from 50 mM l-glutamate in a flow-through system under conditions of pH 4.0 and 55 °C.
authors.affiliation: a Department of Smart-Biohealth , Dong-Eui University , Busan , Republic of Korea.
authors.forename: Ji-Yeon
authors.initials: JY
authors.lastname: Lee
authors.forename: Sung-Jong
authors.initials: SJ
authors.lastname: Jeon
mesh_terms: D000645:Ammonium Sulfate / Q000494:pharmacology; D019149:Bioreactors / Q000382:microbiology; D002614:Chelating Agents / Q000737:chemistry*; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D005968:Glutamate Decarboxylase / Q000737:chemistry* / Q000378:metabolism*; D052196:Levilactobacillus brevis / Q000201:enzymology* / Q000378:metabolism; D009532:Nickel / Q000737:chemistry*; D012685:Sepharose / Q000737:chemistry*; D005680:gamma-Aminobutyric Acid / Q000096:biosynthesis*
chemical_list: D002614:Chelating Agents; D004800:Enzymes, Immobilized; D005680:gamma-Aminobutyric Acid; D009532:Nickel; D012685:Sepharose; D005968:Glutamate Decarboxylase; D000645:Ammonium Sulfate
keywords: bioreactor; glutamate decarboxylase (GAD); immobilization; metal affinity resin; γ-aminobutyric acid (GABA)