• Journal of Applied Biological Chemistry
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• v.56 no.2
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• pp.89-93
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• 2013

Gamma-aminobutyric acid (GABA) aminotransferase (gabT) and succinic semialdehyde dehydrogenase (gabD) genes from Pseudomonas fluorescens KCCM 12537 were cloned into a single pETDuet-1 vector and co-expressed in Escherichia coli BL21(DE3) simultaneously. The mixture of both enzymes, called GABase, is the key enzyme for the enzymatic analysis of GABA. The molecular mass of the GABA aminotransferase and succinic semialdehyde dehydrogenase were determined to be 52.8 and 46.7 kDa following computations performed with the pI/Mw program, respectively. The GABase activity between pH 6.0 and 9.0 for 24 h at $4^{\circ}C$ remained over 75%, but under pH 6.0 decreased rapidly. The GABase activity between 25 and $35^{\circ}C$ by the treatment at pH 8.6 for 30 min remained over 80%, but over $35^{\circ}C$ decreased rapidly. When the activity against GABA was defined as 100%, the purified GABase activity against 5-aminovaleric acid having a similar structure to GABA showed 47.7% and GABase activity against ${\beta}$-alanine, ${\varepsilon}$-amino-n-caproic acid, $_L$-ornithine, $_L$-lysine, and $_L$-aspartic acid showed between 0.3 to 2.3%. The GABA content was analyzed with this co-expressed GABase, compared with the other GABase which was available commercially. As a result, the content of GABA extracted from brown rice, dark brown rice, and black rice were $26.4{\pm}3.5$, $40.5{\pm}4.7$ and $94.7{\pm}9.3{\mu}g/g$, which were similar data of other GABase in the error ranges.

• Journal of Microbiology and Biotechnology
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• v.26 no.4
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• pp.710-716
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• 2016

Gamma-aminobutyric acid (GABA) is a non-protein amino acid, which is an important inhibitor of neurotransmission in the human brain. GABA is also used as the precursor of biopolymer Nylon-4 production. In this study, the carbon flux from the tricarboxylic acid cycle was directed to the GABA shunt pathway for the production of GABA from glucose. The GABA shunt enzymes succinate-semialdehyde dehydrogenase (GabD) and GABA aminotransferase (GabT) were co-localized along with the GABA transporter (GadC) by using a synthetic scaffold complex. The co-localized enzyme scaffold complex produced 0.71 g/l of GABA from 10 g/l of glucose. Inactivation of competing metabolic pathways in mutant E. coli strains XBM1 and XBM6 increased GABA production 13% to reach 0.80 g/l GABA by the enzymes co-localized and expressed in the mutant strains. The recombinant E. coli system developed in this study demonstrated the possibility of the pathway of the GABA shunt as a novel GABA production pathway.

• KSBB Journal
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• v.27 no.2
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• pp.127-130
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• 2012

In this study, the effects of pH, temperature, IPTG concentration and substrate (MSG) concentration on gamma-aminobutyric acid (GABA) production in engineered Escherichia coli were investigated. Glutamate decarboxylase and glutamate/GABA antiporter were overexpressed in GABA aminotransferase knock-out strain for GABA production. The result of optimization study showed the GABA bioconversion was optimized at pH 3.5, $30^{\circ}C$, 0.5 mM IPTG, 10 g/L MSG. At this condition, 5.23 g/L of final GABA concentration of was achieved from 10 g/L of MSG, which corresponded to a GABA yield of 85.77%.

• Korean Journal of Poultry Science
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• v.49 no.4
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• pp.247-253
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• 2022

This study was conducted to investigate the effects of γ-aminobutyric acid (GABA) and a probiotic mixture on egg quality, blood parameters, and corticosterone content in broiler breeder stock in summer in Korea. A total of 72 broiler breeders (Arbor Acres) at 49 weeks old were randomly divided into the following three groups of 24 heads each: control, GABA, and probiotics (1 × 10⁸/g Bacillus licheniformis, 1 × 10⁷/g Lactiplantibacillus plantaru, and 1 × 10⁷/g Clostridium butyricum). The hens were fed a diet containing 50 mg of GABA/kg of feed and 0.1% probiotics for 6 weeks. Egg quality, blood parameters, and stress levels (corticosterone concentration) of the broiler breeders were evaluated. No significant differences were observed in the egg weight, albumen height, Haugh units, yolk color, shell thickness, or shell strength between the groups. There were no significant between-group differences in the blood biochemical profile, including total cholesterol, triglyceride, glucose, total protein, aspartate aminotransferase, alanine aminotransferase, albumin, and inorganic phosphorus concentrations. The corticosterone concentration was significantly lower in the GABA and probiotics groups than the control group (P < 0.05) at 6 weeks of age. In conclusion, these results suggest that dietary probiotics and GABA significantly improve the stress levels of broiler breeders in summer in Korea.

• Korean Journal of Poultry Science
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• v.49 no.4
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• pp.239-246
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• 2022

This study was conducted to investigate the effects of dietary γ-aminobutyric acid (GABA) and a probiotic mixture on egg production and quality, blood parameters, and stress levels (corticosterone) in Hy-Line parent stock during summer in Korea. A total of 105 Hy-Line parent stock aged 24 weeks were randomly divided into three groups, each containing thirty-five birds: control, γ-aminobutyric acid (GABA), and probiotics (1 × 10⁸/g Bacillus licheniformis, 1 × 10⁷/g Lactobacillus plantarum, and 1 × 10⁷/g Corynebacterium butyricum). The hens were fed a diet containing 50 ppm GABA or 0.1% probiotics for 6 weeks. Compared with the control group, the hen-day egg production, egg mass, and feed conversion ratio over the total period were significantly higher in the probiotic group (P<0.05). In contrast no significant differences were detected among groups with respect to egg weight, albumen height, Haugh units, yolk color, shell thickness or shell strength. Similarly, no significant difference were observed among groups with regards to biochemical profile (total cholesterol, triglyceride, glucose, total protein, aspartate aminotransferase, alanine aminotransferase, albumin, and inorganic phosphorus). However, compared with the control group, we did detect significant reductions in corticosterone levels in the GABA and probiotics groups (P<0.05). On the basis of our findings in this study, it would appear that dietary GABA and probiotics can alleviate heat stress in Hy-Line parent stock, with probiotics in particular being found to promote significant improvements in the hen-day egg production, egg mass, and feed conversion of laying hens during the summer season in Korea.

• Korean Journal of Microbiology
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• v.31 no.3
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• pp.224-229
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• 1993

GABA transminase (4-aminobutyrate aminotransferase), which catalyzes the breakdown of the major inhibitory neurotransmitter, GABA, in mammalian brain, was inactivated by preincubation with the mycotoxin patulin. The time course of the reaction was significantly affected by the substrate .alpha.-ketoglutarate, which aforded complete protection against the loss of catalytic activity. The recovery from the inhibition of patulin by the addition of dithiothreitol (DTT) supports that patulin reacts with the sulfhydryl residue in the catalytic domain of the enzyme. The reconstitution of the reduced enzyme and apoenzyme with pyridoxal-5-P(PLP) was inhibited by another mycotoxin, penicilic acid. This mycotoxin may interact with lysyl residue of the enzyme. Therefore, it is postulated that the critical sulfhydryl and lysyl residues in the catalytic domain of the enzyme react with mycotoxin patulin and penicillic acid, respectively.