• Title/Summary/Keyword: GABA production

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Co-Localization of GABA Shunt Enzymes for the Efficient Production of Gamma-Aminobutyric Acid via GABA Shunt Pathway in Escherichia coli

  • Pham, Van Dung;Somasundaram, Sivachandiran;Park, Si Jae;Lee, Seung Hwan;Hong, Soon Ho
    • 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.

Enhanced Production of Gamma-Aminobutyric Acid by Optimizing Culture Conditions of Lactobacillus brevis HYE1 Isolated from Kimchi, a Korean Fermented Food

  • Lim, Hee Seon;Cha, In-Tae;Roh, Seong Woon;Shin, Hae-Hun;Seo, Myung-Ji
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.450-459
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    • 2017
  • This study evaluated the effects of culture conditions, including carbon and nitrogen sources, L-monosodium glutamate (MSG), and initial pH, on gamma-aminobutyric acid (GABA) production by Lactobacillus brevis HYE1 isolated from kimchi, a Korean traditional fermented food. L. brevis HYE1 was screened by the production analysis of GABA and genetic analysis of the glutamate decarboxylase gene, resulting in 14.64 mM GABA after 48 h of cultivation in MRS medium containing 1% (w/v) MSG. In order to increase GABA production by L. brevis HYE1, the effects of carbon and nitrogen sources on GABA production were preliminarily investigated via one-factor-at-a-time optimization strategy. As the results, 2% maltose and 3% tryptone were determined to produce 17.93 mM GABA in modified MRS medium with 1% (w/v) MSG. In addition, the optimal MSG concentration and initial pH were determined to be 1% and 5.0, respectively, resulting in production of 18.97 mM GABA. Thereafter, response surface methodology (RSM) was applied to determine the optimal conditions of the above four factors. The results indicate that pH was the most significant factor for GABA production. The optimal culture conditions for maximum GABA production were also determined to be 2.14% (w/v) maltose, 4.01% (w/v) tryptone, 2.38% (w/v) MSG, and an initial pH of 4.74. In these conditions, GABA production by L. brevis HYE1 was predicted to be 21.44 mM using the RSM model. The experiment was performed under these optimized conditions, resulting in GABA production of 18.76 mM. These results show that the predicted and experimental values of GABA production are in good agreement.

Production of Fermented Saccharina Japonica Extract with Enhanced GABA Content (GABA 함량이 강화된 발효다시마 추출액 제조)

  • Hur, Sun-Sun
    • Journal of the Korean Applied Science and Technology
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    • v.39 no.4
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    • pp.517-526
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    • 2022
  • The purpose of this study was to enhance the gamma-aminobutyric acid (GABA) production of sea tangle extracts, through techniques based on enzymatic hydrolysis and the addition of mixed fermentative lactic acid bacteria. GABA production in the strains was qualitatively confirmed via detection of colored spots using thin layer chromatography. L. plantarum KCTC 21004, L. acidophilus KCTC 3164 and L. sakei subsp. sakei KCTC 3598 were selected as the suitable strains for GABA production. As for the characteristics of fermentation of lactic acid bacteria using the selected strain, as the fermentation time increased, the titrated acidity increased and the pH showed a tendency to decrease. Among the three strains with excellent GABA production ability, L. plantarum KCTC 21004 showed excellent GABA production of 136.4 mg/100g. These research results are expected to be provided a basis for the utilization of lactic acid bacteria in GABA production using a sea tangle extract.

Characterization of the Production of Biogenic Amines and Gamma-Aminobutyric Acid in the Soybean Pastes Fermented by Aspergillus oryzae and Lactobacillus brevis

  • Kim, Nam Yeun;Ji, Geun Eog
    • Journal of Microbiology and Biotechnology
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    • v.25 no.4
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    • pp.464-468
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    • 2015
  • The production of gamma-aminobutyric acid (GABA) using GABA-producing lactic acid bacteria (LAB) has been considered to be an attractive strategy. However, some LAB may produce biogenic amines (BA), which may be of concern from the safety viewpoint. The aim of the present study was to characterize the production of GABA and BA in the soybean pastes fermented by Aspergillus oryzae (A. oryzae) FMB S46471 and GABA-producing Lactobacillus brevis (L. brevis) GABA 100. After a ripening period of 90 days, the levels of BA (putrescine, cadaverine, histamine, and tyramine) and GABA in the fermented soybean were assessed by highperformance liquid chromatography. The soybean pastes fermented by A. oryzae and L. brevis showed a range of 7,130-11,592 mg/kg for GABA, 178-305 mg/kg for tyramine, 139-163 mg/kg for putrescine, 7.4-10.8 mg/kg for histamine, and 7.1-7.9 mg/kg for cadaverine, whereas the soybean pastes fermented by A. oryzae only showed a range of 30-1,671 mg/kg for GABA, 0.8-189 mg/kg for tyramine, 1.3-85 mg/kg for putrescine, up to 3.6 mg/kg for histamine, and 0.2-2.4 mg/kg for cadaverine. The results showed that the production of GABA was accompanied by the increase in the production of BA, even though the production levels of histamine and cadaverine were very low. This is the first study to simultaneously characterize the production of BA and GABA in GABA-enriched fermented soybean pastes, and warrants further study to minimize the production of BA while optimizing the production of GABA.

Effects and Optimization of Gamma-Amino Butyric Acid (GABA) Production Process using Glutamate Decarboxylase (GAD) (Glutamate Decarboxylase (GAD)를 이용한 Gamma-Amino Butyric Acid (GABA) 생산 및 최적화)

  • Kim, Eui Jin;Lee, Jung-Heon
    • KSBB Journal
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    • v.29 no.6
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    • pp.426-431
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    • 2014
  • pH controlled batch reactor and bubble column reactors have been developed in this research. They were used to produce high concentration of GABA and to determine optimal pH for GABA production. Glutamate decarboxylase (GAD) was isolated from recombinant E. coli and used for GABA production from monosodium glutamate (MSG). pH control was inevitable because the pH increased with MSG consumption. GAD showed highest activity at acidic conditions at pH 5.5 but the optimal pH for GABA production was pH 6.0. When 1.5 mole of MSG was used as reactant, the 1.05 mole of GABA was produced after 10 hrs batch reaction. Using bubble column reactors, 80 % of MSG was converted to GABA for 6 hrs reaction and 1.2 mole of GABA was produced.

Probiotic Properties and Optimization of Gamma-Aminobutyric Acid Production by Lactiplantibacillus plantarum FBT215

  • Kim, Jaegon;Lee, Myung-Hyun;Kim, Min-Sun;Kim, Gyeong-Hwuii;Yoon, Sung-Sik
    • Journal of Microbiology and Biotechnology
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    • v.32 no.6
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    • pp.783-791
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    • 2022
  • Gamma-aminobutyric acid (GABA) improves various physiological illnesses, including diabetes, hypertension, depression, memory lapse, and insomnia in humans. Therefore, interest in the commercial production of GABA is steadily increasing. Lactic acid bacteria (LAB) have widely been reported as a GABA producer and are safe for human consumption. In this study, GABA-producing LAB were preliminarily identified and quantified via GABase assay. The acid and bile tolerance of the L. plantarum FBT215 strain were evaluated. The one-factor-at-a-time (OFAT) strategy was applied to determine the optimal conditions for GABA production using HPLC. Response surface methodology (RSM) with Box-Behnken design was used to predict the optimum GABA production. The strain FBT215 was shown to be acid and bile tolerant. The optimization of GABA production via the OFAT strategy resulted in an average GABA concentration of 1688.65 ± 14.29 ㎍/ml, while it was 1812.16 ± 23.16 ㎍/ml when RSM was applied. In conclusion, this study provides the optimum culture conditions for GABA production by the strain FBT215 and indicates that L. plantarum FBT215 is potentially promising for commercial functional probiotics with health claims.

Production of GABA (gamma amino butyric acid) by Lactic Acid Bacteria

  • Kook, Moo-Chang;Cho, Seok-Cheol
    • Food Science of Animal Resources
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    • v.33 no.3
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    • pp.377-389
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    • 2013
  • Gamma-amino butyric acid (GABA) is a kind of pharmacological and biological component and its application is wide and useful in Korea specially, becoming aging society in the near feature. GABA is request special dose for the purposed biological effect but the production of concentrated GABA is very difficult due to low concentration of glutamic acid existed in the fermentation broth. To increase GABA concentrate using fermentation technology, high content of glutamic acid is required. For this reason, various strains which have the glutamic acid decarboxylase (GAD) and can convert glutamic acid to GABA, were isolated from various fermented foods. Most of GABA producing strains are lactic acid bacteria isolated from kimchi, especially added monosodium glutamate (MSG) as a taste enhancer. Optimizing the formulation of culture media and the culture condition, GABA conversion yield and amounts were increased. Finally GABA concentration of fermentation broth in batch or fed batch fermentation reached 660 mM or 1000 mM, respectively. Furthermore formulation of culture media for GABA production developed commercially. Many studies about GABA-rich product have been continued, so GABA-rich kimchi, cheese, yogurt, black raspberry juice and tomato juices has been also developed. In Korea many biological effects of GABA are evaluated recently and GABA will be expected to be used in multipurpose.

Production of γ-aminobutyric Acid Using the Korean Hull-less Barley Bran with Glutamate (국내산 쌀보리 맥강을 이용한 glutamate로부터 GABA 생산)

  • Lee, Mi Ja;Kim, Shin Hye;Kim, Hyung Soon;Kim, Hyun Young;Seo, Woo Duck;Choi, Sik Won;Lee, Kwang Sik;Jang, Ki Chang
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.63 no.1
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    • pp.35-40
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    • 2018
  • ${\gamma}$-Aminobutyric acid (GABA) has antihypertensive and anti-stress effects on humans. The present study aimed to investigate the effect of glutamate concentration, bran content, and barley cultivars on GABA production and determine the optimal reaction condition. Barley bran resulted in higher GABA productivity compared with that of rice bran. The higher the bran content, the higher was the GABA productivity. Furthermore, high glutamate concentration resulted in high GABA production. However, there was a decrease in the glutamate conversion rate. The production of GABA varied with temperature and barley cultivar. The optimal condition for GABA production using barley bran was 0.15 g/mL barley content, 10 mM glutamate concentration, and $20^{\circ}C$ reaction temperature. Under optimal condition, the GABA concentration was 10.34 mM, and glutamate conversion rate was 75.1%. Furthermore, the GABA productivity of the GABA production reaction using barley bran differed depending on the cultivar. Dasong and Yeongbaekchal showed higher GABA productivity than that by other cultivars.

Production of gamma-Aminobutyric Acid (GABA) by Lactobacillus plantarum subsp. plantarum B-134 Isolated from Makgeolli, Traditional Korean Rice Wine (한국전통주인 막걸리로부터 분리한 Lactobacillus plantarum subsp. plantarum B-134의 gamma-aminobutyric acid (GABA)의 생산)

  • Lee, Hyun-Ju;Son, Jae-Young;Lee, Sang-Jae;Lee, Han-Seung;Lee, Bae-Jin;Choi, In-Soon;Sohn, Jae Hak
    • Journal of Life Science
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    • v.27 no.5
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    • pp.567-574
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    • 2017
  • This study is to isolate and identify ${\gamma}$-amino butyric acid (GABA) producing lactic acid bacteria (LAB) from Makgeolii, traditional Korean rice wine and then establish the optimal culture conditions for GABA production. Sixty four LAB from Makgeolli were isolated according to the characteristics of the shape and color of the colony grown on MRS agar plate. The GABA production of the isolated strain cultured in MRS broth contained 1% MSG (mono-sodium glutamate) were determined and evaluated by TLC and HPLC analysis. Strain B-134 was selected for highest GABA production. From the analysis of 16S rRNA and glutamate decarboxylase B (gadB) gene sequences, strain B-134 was tentatively identified as a Lactobacillus plantarum subsp. plantarum B-134. Effects of culture parameters, including glutamic acid level, culture temperature, NaCl level, and pH on GABA production were investigated for culture optimization. The optimum culture condition for GABA production by B-134 were culture temperature of $37^{\circ}C$, pH of 5.7, NaCl content of 0% (w/v) and MSG content of 3% (w/v), which produced 25 mM of GABA during cultivation time of 48 hr. From these results, strain B-134 is expected to be utilized as useful microorganisms for GABA-enriched health beneficial food.

Optimal Conditions for the Production of Gamma-aminobutyric Acid by Enterococcus casseliflavus PL05 Isolated from Oenanthe javanica

  • Choi, Se Mi;Kim, Jeong A;Kim, Geun Su;Kwon, Do Young;Kim, Sang Gu;Lee, Sang yun;Lee, Kang Wook
    • Journal of Food Hygiene and Safety
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    • v.37 no.1
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    • pp.21-28
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    • 2022
  • In this study, a new lactic acid bacterium (LAB) that could produce gamma-aminobutyric acid (GABA) was isolated from Oenanthe javanica (water celery) and identified as an Enteroccoccus casseliflavus strain. Until recently, there have been many studies on the gamma-aminobutyric acid producing lactic acid bacterium, as well as on some lactic acid bacterium in Enteroococcs genus, but none on the species E. casseliflavus. Therefore, in the purpose of finding the optimal conditions for GABA production of E. casseliflavus PL05, the effects of several conditions including the type of mediums, growth temperatures, initial pH, growth time, L-mono sodium glutamate (MSG) concentration, and carbon source were tested. The study revealed that the PL05 strain grew better in the Brain Heart Infusion (BHI) medium than in the Man, Rogosa, and Sharpe (MRS) or Tryptic Soy Broth (TSB) medium. Also, similar results were obtained with GABA production conditions. As a result of analysis on the GABA production yield by concentration of MSG, a GABA substrate, the highest production was found at 7% of MSG concentration. However, since similar level of production was found at 5%, it is considered to be more efficient to use 5% MSG concentration. The analysis on the growth and GABA production yield by carbon sources showed the highest results when maltose was used. From the final test under the optimal conditions found, 140.06±0.71 mM of GABA was produced over 24 hours with the conversion rate of 78.95%. Lastly, from the sensitivity analysis on the 10 different antibiotics, including vancomycin, it was found that there were not confirmed cases of resistance.