• Title/Summary/Keyword: glutamate-producing bacteria

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Production of L-Ornithine by Citrulline Auxotrophic Mutants of Glutamate-Producing Bacteria

  • CHOI, DAE KEON;WUK SANG RYU;BONG HYUN CHUNG;SOO WAN NAM;YOUNG HOON PARK
    • Journal of Microbiology and Biotechnology
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    • v.2 no.2
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    • pp.102-107
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    • 1992
  • For the purpose of producing L-ornithine by microbial fermentation, mutant strains were developed from glutamate-producing bacteria by mutagenesis using N-methyl-N'-nitro-N-nitrosoguanidine (NTG) and UV irradiation. Brevibacterium ketoglutamicum BK1046, a L-citrulline auxotroph which is also resistant to arginine hydroxamate (Arghx), was isolated and selected as the best producer of L-ornithine. This strain was capable of producing L-ornithine at a concentration of 24 g/l after 69 hours of cultivation in the 21 jar fermentor. The optimum supplementary level of L-arginine, a substitute for L-citrulline, was found to be about 0.2 g/l in the shake-flask fermentation.

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Isolation and Characterization of Lactobacillus buchneri Strains with High ${\gamma}$-Aminobutyric Acid Producing Capacity from Naturally Aged Cheese

  • Park, Ki-Bum;Oh, Suk-Heung
    • Food Science and Biotechnology
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    • v.15 no.1
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    • pp.86-90
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    • 2006
  • Two lactic acid bacteria (LAB) with high ${\gamma}$-aminobutyric acid (GABA)-producing capacity were isolated from naturally aged cheese. Examination of the biochemical features using an API kit indicated that the two strains belonged to Lactobacillus. They were gram positive, rod-type bacteria, and fermented arabinose, melezitose, melibiose and xylose, but did not utilize cellobiose or trehalose. 16S rDNA sequencing analysis confirmed that they were Lactobacillus buchneri and Lactobacillus sp. They were accordingly named as Lactobacillus buchneri OPM-1 and Lactobacillus sp. OPM-2, and could produce GABA from MRS broth supplemented with 10 g/L of monosodium glutamate (MSG) at a productivity of 91.7 and 116.7 mg/L/hr, respectively. Cell extracts of L. buchneri OPM-1 and Lactobacillus sp. OPM-2 showed glutamate decarboxylase (GAD) activity, for which the optimum pH and temperature were 5.5 and $30^{\circ}C$, respectively.

Characteristics of Potential Gamma-Aminobutyric Acid-Producing Bacteria Isolated from Korean and Vietnamese Fermented Fish Products

  • Vo, Thi Thu-Thao;Park, Jong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.29 no.2
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    • pp.209-221
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    • 2019
  • Gamma-aminobutyric acid (GABA) is a neurotransmitter that exerts several physiological functions and positive effects on human health. The aim of this study was to isolate and characterize the strains that had GABA-producing abilities from various fermented fish products. A total of 91 acid-producing strains were isolated from 41 samples of fermented fish products, and 27 strains showing GABA-producing abilities were identified by the 16S rDNA sequences. Among the strains, 31% strains tolerated at high-salt environment of 10-20% throughout the fermentation of fish sauces. The 27 isolates that produced GABA at various concentrations did so in the range of 5 to 454 mM. These GABA-producing isolates were identified as lactic acid bacteria of 14 strains, which included twelve Lactococcus lactis, one Enterococcus faecium, and one Lactococcus pentosus; eight Bacillus cereus group, which included seven B. thuringiensis and one B. cereus; and five Staphylococcus spp. Interestingly, with Vietnamese fish sauces, we mostly identified species of B. thuringiensis and Staphylococcus spp., while with Korean fermented fish products, the majority of the strains identified belonged to L. lactis. Among the strains, B. thuringiensis LH2134 produced the highest levels of GABA at 366 mM among the strains identified from Vietnamese fish sauces, whereas L. lactis LA43, a new strain isolated from Korean jeotgal (salted shrimp paste), produced the highest amount of GABA at 454 mM and the glutamate concentration in the medium was essential for GABA accumulation. Therefore, such the isolates might serve as good starters for development of more GABA-reinforced foods among fermented fish products.

The Role of Glutamic Acid-producing Microorganisms in Rumen Microbial Ecosystems (반추위 미생물생태계에서의 글루탐산을 생성하는 미생물의 역할)

  • Mamuad, Lovelia L.;Lee, Sang-Suk
    • Journal of Life Science
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    • v.31 no.5
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    • pp.520-526
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    • 2021
  • Microbial protein is one of the sources of protein in the rumen and can also be the source of glutamate production. Glutamic acid is used as fuel in the metabolic reaction in the body and the synthesis of all proteins for muscle and other cell components, and it is essential for proper immune function. Moreover, it is used as a surfactant, buffer, chelating agent, flavor enhancer, and culture medium, as well as in agriculture for such things as growth supplements. Glutamic acid is a substrate in the bioproduction of gamma-aminobutyric acid (GABA). This review provides insights into the role of glutamic acid and glutamic acid-producing microorganisms that contain the glutamate decarboxylase gene. These glutamic acid-producing microorganisms could be used in producing GABA, which has been known to regulate body temperature, increase DM intake and milk production, and improve milk composition. Most of these glutamic acid and GABA-producing microorganisms are lactic acid-producing bacteria (LAB), such as the Lactococcus, Lactobacillus, Enterococcus, and Streptococcus species. Through GABA synthesis, succinate can be produced. With the help of succinate dehydrogenase, propionate, and other metabolites can be produced from succinate. Furthermore, clostridia, such as Clostridium tetanomorphum and anaerobic micrococci, ferment glutamate and form acetate and butyrate during fermentation. Propionate and other metabolites can provide energy through conversion to blood glucose in the liver that is needed for the mammary system to produce lactose and live weight gain. Hence, health status and growth rates in ruminants can be improved through the use of these glutamic acid and/or GABA-producing microorganisms.

Studies on the Bacterial Production of L-Glutamate from Acetate Part I. Screening and Identification of L-Glutamate Producing Bacteria. (초산을 이용한 글루타민산의 발효생산에 관한 연구 제 1보 글루타민산 생산균주의 분리 및 동정)

  • 하덕모;노완섭
    • Microbiology and Biotechnology Letters
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    • v.2 no.2
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    • pp.103-109
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    • 1974
  • In the cource of the studies on L-glutamic acid production from acetic acid, 383 strains capable of assimilating acetate as sole source of carbon were isolated from 279 kinds of soil sample. Out of them, 5 strains which produced relatively larger amount of L-glutamate from acetate were selected and named Brevibacterium flavum nov. sp. D1005B, Corynebacterium glutamicum nov. sp. D1025A, Brevib. flavum nov. sp. D2209B, Coryneb. acetoacidophilum nov. sp. D2212B and Coryneb. acetoacidophilum nov. sp. D2349A respectively.

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Characterization of a Glutamate Decarboxylase (GAD) from Enterococcus avium M5 Isolated from Jeotgal, a Korean Fermented Seafood

  • Lee, Kang Wook;Shim, Jae Min;Yao, Zhuang;Kim, Jeong A;Kim, Hyun-Jin;Kim, Jeong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.27 no.7
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    • pp.1216-1222
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    • 2017
  • To develop starters for the production of functional foods or materials, lactic acid bacteria producing ${\gamma}-aminobutyric$ acid (GABA) were screened from jeotgals, Korean fermented seafoods. One isolate producing a high amount of GABA from monosodium $\text\tiny{L}$-glutamate (MSG) was identified as Enterococcus avium by 16S rRNA gene sequencing. E. avium M5 produced $18.47{\pm}1.26mg/ml$ GABA when incubated for 48 h at $37^{\circ}C$ in MRS broth with MSG (3% (w/v)). A gadB gene encoding glutamate decarboxylase (GAD) was cloned and overexpressed in E. coli BL21 (DE3) using the pET26b (+) expression vector. Recombinant GAD was purified through a Ni-NTA column and the size was estimated to be 53 kDa by SDS-PAGE. Maximum GAD activity was observed at pH 4.5 and $55^{\circ}C$and the activity was dependent on pyridoxal 5'-phosphate. The $K_m$ and $V_{max}$ values of GAD were $3.26{\pm}0.21mM$ and $0.0120{\pm}0.0001mM/min$, respectively, when MSG was used as a substrate. Enterococcus avium M5 secretes a lot of GABA when grown on MRS with MSG, and the strain is useful for the production of fermented foods containing a high amount of GABA.

Isolation of Novel Pseudomonas diminuta KAC-1 Strain Producing Glutaryl 7-Aminocephalosporanic Acid Acylase

  • Kim, Dae-Weon;Kang, Sang-Mo;Yoon, Ki-Hong
    • Journal of Microbiology
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    • v.37 no.4
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    • pp.200-205
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    • 1999
  • 7-Aminocephalosporanic acid (7-ACA) is the initial compound in preparation of cephalosporin antibiotics widely used in clinical treatment. Bacteria producing glutaryl 7-ACA acylase, which convert cephalosporin C to 7-ACA, has been screened in soil samples. A bacterial strain exhibiting high glutaryl 7-ACA acylase activity, designated KAC-1, was isolated and identified as a strain of Pseudomonas diminuta by characterizing its morphological and physiological properties. The screening procedures include culturing on enrichment media containing glutaric acid, glutamate, and glutaryl 7-aminocephalosporanic acid as selective carbon sources. To enhance enzyme production, optimal cultivation conditions were investigated. This strain grew optimally at pH 7 to 9 and in temperatures of 20 to 40 C, but acylase production was higher when the strain was grown at 25 C. Glutaric acid, glutamate and glucos also acted as inducers for acylase production. In a jar fermenter culture, P. diminuta KAC-1 produce acylase in a growth-associated manner. The substrate specificity of KAC-1 acylase by cell extract showed that this enzyme had specificity toward glutaryl 7-ACA, glutaryl 7-ADCA, but not cephalosporin C.

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Studies on the Alkalophilic Bacteria Producing Alkaline Protease and its Enzyme Activities (알칼리성 Protease를 생성하는 알칼리성 세균 및 그 효소활성에 관한 연구)

  • Park, Soo-Jin;Lee, Kang-Man;Bae, Moo
    • YAKHAK HOEJI
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    • v.34 no.1
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    • pp.47-53
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    • 1990
  • Alkalophilic bacteria isolated from compost were selected, identified and tested for production of alkaline protease. The bacterium was tentatively assigned to Bacillus sp. based on the morphological, cultural and biochemical characteristics. The optimum pH of growth was 10 Galactose and Sodium glutamate enhanced the production of alkaline protease. The protease was most active at pH 11.0 and $60^{\circ}C$ and stable in the range of pH 5-11 and temp. $30^{\circ}-55^{\circ}C$. The protease was stabilized by the presence of calcium salts.

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Production and Characterization of GABA Rice Yogurt

  • Park, Ki-Bum;Oh, Suk-Heung
    • Food Science and Biotechnology
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    • v.14 no.4
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    • pp.518-522
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    • 2005
  • Yogurt containing high ${\gamma}$-aminobutyric acid (GABA) was developed using lactic acid bacteria and germinated brown rice. Lactobacillus acidophilus, L. plantarum, and L. brevis OPY-1 strains were inoculated into Lactobacillus MRS broth for use as yogurt starter. After treatment with 5% monosodium glutamate in MRS broth, L. brevis OPY-1 strain isolated from Kimchi produced GABA concentration of 8,003.28 nmol/mL. Starter was inoculated into fermentation substrate mixture containing germinated brown rice extract and blend of powdered whole milk and skim milk. Samples were incubated, and viable cell colonies were counted. Highest number of lactic acid bacteria was reached between 16 and 20 hr. Concentrated rice milk fermented with high GABA-producing strain contained GABA concentrations of $137.17\;{\mu}g/g$ D.W., whereas concentrated fermented milk prepared by conventional method contained GABA of $1.29\;{\mu}g/g$ D.W. Sensory evaluation panelists gave favorable ratings to fermented rice milk containing high GABA concentration.

Gamma-Aminobutyric Acid Production from a Novel Enterococcus avium JS-N6B4 Strain Isolated from Edible Insects

  • Jo, Min-Ho;Hong, Seong-Jin;Lee, Ha-Nul;Ju, Jung-Hyun;Park, Bo-Ram;Lee, Jun-ho;Kim, Sun-Am;Eun, Jong-Bang;Wee, Young-Jung;Kim, Young-Min
    • Journal of Microbiology and Biotechnology
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    • v.29 no.6
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    • pp.933-943
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    • 2019
  • Gamma-aminobutyric acid (GABA)-producing strains were isolated from four edible insects and subjected to 16S rRNA sequence analysis. Among the four GABA-producing bacteria, Enterococcus avium JS-N6B4 exhibited the highest GABA-production, while cultivation temperature, initial pH, aerobic condition, and mono-sodium glutamate (MSG) feeding were found to be the key factors affecting GABA production rate. The culture condition was optimized in terms of glucose, yeast extract, and MSG concentrations using response surface methodology (RSM). GABA production up to 16.64 g/l was obtained under the conditions of 7 g/l glucose, 45 g/l yeast extract, and 62 g/l MSG through the optimization of medium composition by RSM. Experimental GABA production was 13.68 g/l, which was close to the predicted value (16.64 g/l) calculated from the analysis of variance, and 2.79-fold higher than the production achieved with basic medium. Therefore, GABA-producing strains may help improve the GABA production in edible insects, and provide a new approach to the use of edible insects as effective food biomaterials.