Purification and Characterization of Glutamate Decarboxylase from<i>Lactobadllus bvevis</i>IFO 12005

Ueno, Yukihiko; Hayakawa, Kiyoshi; Takahashi, Saori; Oda, Kōhei

doi:10.1271/bbb.61.1168

Cited by 154 publications

(104 citation statements)

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“…paracasei and Lb. brevis), GAD activity was still observed at pH 4.0, 18) but very low levels of GAD activity have been observed at pH 4.0 in a low GABA-producing strain (L. lactis). 25) These results suggest that low-pH GAD activity might be important for producing high levels of GABA in LAB.…”

Section: Discussionmentioning

confidence: 93%

“…brevis. 18) In Lactobacillus species, the dimer formation of GAD might be conserved, whereas the GAD of E. coli forms a hexamer. 24) The optimum temperature for the GAD activity of Lb.…”

Section: Discussionmentioning

confidence: 99%

“…paracasei differed from that previously reported for GAD from other LAB. 18,20) The enzyme was active within a broad pH range as compared with GADs from other LAB species.…”

Section: Properties Of Gad In Lb Paracaseimentioning

confidence: 95%

“…17) The purification and enzymatic properties of GAD in such LAB species as Lactobacillus and Lactococcus have been reported. Ueno et al 18) purified GAD from Lb. brevis and then determined its biochemical characteristics.…”

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confidence: 99%

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Characterization of Glutamate Decarboxylase from a High γ-Aminobutyric Acid (GABA)-Producer,Lactobacillus paracasei

Komatsuzaki

Nakamura

Kojima

et al. 2008

Bioscience, Biotechnology, and Biochemistry

143

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Section: Discussionmentioning

confidence: 93%

“…brevis. 18) In Lactobacillus species, the dimer formation of GAD might be conserved, whereas the GAD of E. coli forms a hexamer. 24) The optimum temperature for the GAD activity of Lb.…”

Section: Discussionmentioning

confidence: 99%

“…paracasei differed from that previously reported for GAD from other LAB. 18,20) The enzyme was active within a broad pH range as compared with GADs from other LAB species.…”

Section: Properties Of Gad In Lb Paracaseimentioning

confidence: 95%

mentioning

confidence: 99%

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Characterization of Glutamate Decarboxylase from a High γ-Aminobutyric Acid (GABA)-Producer,Lactobacillus paracasei

Komatsuzaki

Nakamura

Kojima

et al. 2008

Bioscience, Biotechnology, and Biochemistry

143

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“…Ueno et al (1997) also used a short incubation for the solubilization of GAD from L. brevis by lysozyme.…”

Section: Resultsmentioning

confidence: 99%

Purification and Characterization of Glutamate Decarboxylase from Aspergillus oryzae

Tsuchiya

Nishimura

Iwahara

2003

FSTR

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We purified glutamate decarboxylase (GAD) [EC4.1.1.15] from Aspergillus oryzae and characterized its biochemical and kinetic properties. GAD was purified by ammonium sulfate at 30-70% saturation and chromatographies on Sephacryl S-300, DEAE-FF and CM-FF. The purification of GAD from the crude enzyme solution was 40-fold and the recovery rate was 4.9%. About 230 g of purified enzyme was obtained from 20 g of the mycelia of A. oryzae. The purified preparation of the enzyme showed a single protein band on SDS-PAGE. The molecular weight of purified GAD by SDS-PAGE and gel filtration was estimated to be 48 kDa and 300 kDa, respectively, suggesting that purified GAD had a hexameric structure. The K m value for L-glutamic acid, a substrate of the enzyme, was estimated to be 13 mM. The optimum pH and temperature of GAD were 5.5 and 60˚C, respectively. The GAD activity was stable up to 40˚C.Keywords: glutamate decarboxylase, Aspergillus oryzae, purification, ␥-amino-butyric acid Glutamate decarboxylase (GAD; EC4.1.1.15) produces ␥-amino-butyric acid (GABA) from glutamate. GABA has several physiological effects on the human body, including neurotransmitting, hypotensive and diuretic effects. Several attempts to enrich GABA in functional foods have been reported: GABArich green tea produced by anaerobic treatment of green tea (Ohmori et al., 1987), GABA accumulation in red mold rice (Tsuji et al., 1992), GABA accumulation in rice germ by soaking in water (Saikusa et al., 1994), GABA enrichment in brown rice by high-pressure treatment (Kinefuchi et al., 1999) and GABA production by lactic acid bacteria .We have recently found high GAD activity in Aspergillus oryzae cultured in a liquid medium, and it was specially rich in A. oryzae used for miso and sake brewing (Tsuchiya & Nishimura, 2002a). Furthermore, we also reported that the batch reactor using pellet form mycelium of A. oryzae (for miso production), which was cultured in liquid medium achieved an extremely high GABA production rate (8.5 mmol GABA/g Aspergillus mycelium/h) (Tsuchiya et al., 2002b).GAD is widely distributed in animate nature. In mammals, GAD has been purified from human brain (Blindermann et al., 1978) and mouse brain (Wu et al., 1973). In higher plants, GAD has been purified from squash (Matsumoto et al., 1986). In bacteria, the enzymatic properties of GAD have been reported in both Escherichia coli (Shikuya & Schwert, 1960;Fonda, 1985) and Lactobacillus brevis . Although GAD obtained from A. oryzae has not been purified, the gene was recently cloned and the nucleotide sequence was determined (Iwai et al., 2001). The detailed biochemical nature of the enzyme is unclear, which we believe important in promoting the GABA production in food processing by effective utilization of the GAD from A. oryzae. In this study, we purified GAD from the mycelium of A. oryzae and examined some biochemical characteristics of the enzyme. Materials and MethodsMicroorganism and the culture conditions Aspergillus oryzae used in this study was isolated from tane-koji SR-108...

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Gamma‐Aminobutyric Acid

Jiang

Zhang

2010

Bioactive Proteins and Peptides as Functional Foods and Nutraceuticals

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Purification and Characterization of Glutamate Decarboxylase fromLactobadllus bvevisIFO 12005

Cited by 154 publications

References 0 publications

Characterization of Glutamate Decarboxylase from a High γ-Aminobutyric Acid (GABA)-Producer,Lactobacillus paracasei

Characterization of Glutamate Decarboxylase from a High γ-Aminobutyric Acid (GABA)-Producer,Lactobacillus paracasei

Purification and Characterization of Glutamate Decarboxylase from Aspergillus oryzae

Gamma‐Aminobutyric Acid

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