• 약학회지
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• 제42권3호
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• pp.248-256
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• 1998

We had already reported that we purified N-${\beta}$-alanyl-5-S-glutathionyl-3,4-dihydroxyphenylalanine (5-S-GAD), a novel antibacterial substance from the immunized adult Sarcoph aga peregrina (Flesh fly). We found that the antibacterial activity of synthetic 5-S-GAD is equal to that of authentic 5-S-GAD without a specificity of antibacterial activity against Gram positive and Gram negative. Significant synergism was detected between 5-S-GAD and streptomycin against streptomycin resistant strain E.coli K12 594. It has an antitumor activity against several tumor cell lines at a concentration of $100{\mu}M$. However, no cytotoxic activity against murine macrophage was detected at a concentration of $500{\mu}M$. Furthermore, haemolytic activity against sheep erythrocytes was not detected at the same concentration. We suggest that the S-conjugation of glutathion with dihydroxyphenylalanine might be important to increase antibacterial activity of dihydroxyphenylalanme.

• Journal of Microbiology and Biotechnology
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• 제25권5호
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• pp.696-703
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• 2015

A gamma-aminobutyric acid (GABA)-producing microorganism was isolated from jeot-gal (anchovy), a Korean fermented seafood. The isolate, A156, produced GABA profusely when incubated in MRS broth with monosodium glutamate (3% (w/v)) at 37℃ for 48 h. A156 was identified as Lactobacillus sakei by 16S rRNA gene sequencing. The GABA conversion yield was 86% as determined by GABase enzyme assay. The gadB gene encoding glutamate decarboxylase (GAD) was cloned by PCR. gadC encoding a glutamate/GABA antiporter was located immediately upstream of gadB. The operon structure of gadCB was confirmed by RT-PCR. gadB was overexpressed in Escherichia coli BL21(DE3) and recombinant GAD was purified. The purified GAD was 54.4 kDa in size by SDS-PAGE. Maximum GAD activity was observed at pH 5.0 and 55℃ and the activity was dependent on pyridoxal 5'-phosphate. The K_m and V_max of GAD were 0.045 mM and 0.011 mM/min, respectively, when glutamate was used as the substrate.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1995년도 춘계학술대회
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• pp.73-73
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• 1995

We investigated the effect of derivatized phospholipid, P-pyridoxyl dipalmiuylphosphatidylethanolamine (P-pyr-DPPE), on the catalytic activity of purified porcine brain glutamate decarboxylase(GAD) which catalyzes the synthesis of GABA known as major inhibitory neurotransmitter in CNS. When the P-pyr-DPPE was incorporated into dipalmitdylphosphatidylcholine(DPPC) or phosphatidylserine(PS) vesicles, these vesicles enhanced the catalytic activity of GAD. P-pyr-DPPE also interacted with apoglutamate decarboxylase(apoGAD) and produced the free pyridoxal-5-phosphate(PLP) which is the natural cofactor of GAD. This result indicated that apoGAD catalyzed the cleavage reaction of the P-pyridoxyl moiety of the derivatized phopholipid to generate free PLP, and then free PLP bound to the apoGAD resulting in restroration of the catalytic activity of the enzyme.

• 한국미생물·생명공학회지
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• 제49권1호
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• pp.9-17
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• 2021

A γ-aminobutyric acid (GABA)-producing microorganism was isolated from galchi (hairtail fish, Trichiurus lepturus) jeotgal, a Korean salted and fermented seafood. The G144 isolate produced GABA excessively when incubated in MRS broth containing monosodium glutamate (MSG, 3%, w/v). G144 was identified as Lactobacillus brevis through 16S rRNA and recA gene sequencing. gadB and gadC encoding glutamate decarboxylase (GAD) and glutamate/GABA antiporter, respectively, were cloned and gadB was located downstream of gadC. The operon structure of gadCB was confirmed by reverse transcription (RT)-polymerase chain reaction. gadB was overexpressed in Escherichia coli and recombinant GAD was purified and its size was 54.4 kDa as evidenced by SDS-PAGE results. Maximum GAD activity was observed at pH 5.0 and 40℃ and the activity was dependent on pyridoxal 5'-phophate. The K_m and V_max of GAD were 8.6 mM and 0.01 mM/min, respectively.

• Journal of Microbiology and Biotechnology
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• 제27권7호
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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.

• 한국미생물·생명공학회지
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• 제43권3호
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• pp.300-305
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• 2015

효율적인 γ-aminobutyric acid (GABA)의 생산을 위해 Lactobacillus plantarum WCFS1로부터 글루탐산 탈탄산효소(glutamate decarboxylase, GAD)를 대장균에 발현, 정제 후 silica beads에 covalent coupling 방법을 이용하여 고정화하였다. 고정화된 효소의 특성을 고정화하지 않은 효소와 비교한 결과, 모든 pH의 범위(pH 3.5–6.0)에서 80% 이상의 활성을 나타내었으며 pH 안정성과 열 안정성 모두 증대되었다. 이 고정화 효소를 packed-bed reactor에 충진하여 GABA의 생산성을 확인한 결과 1리터당 1시간에 최대 41.7 g의 GABA 생산이 가능한 것으로 확인되었다.

• 한국미생물·생명공학회지
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• 제50권3호
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• pp.319-327
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• 2022

γ-Aminobutyric acid (GABA) is a multi-functional compound with broad applications for food industry. GABA producing bacteria were isolated from cabbage kimchi. Among them, B737 was the best GABA producer when culture supernatants were analyzed by TLC. B737 was identified as Levilactobacillus brevis by 16S rRNA gene sequencing. Its glutamate decarboxylase (GAD) gene was cloned by PCR and the nucleotide sequence determined. B737 GAD consisting of 485 amino acids is the largest in size among GADs reported from LAB so far. gadB from L. brevis B737 was overexpressed in Escherichia. coli BL21(DE3) using pET26b(+).pET26b(+). The recombinant GAD was purified and its size was 55 kDa by SDS-PAGE. Maximum GAD activity was observed at pH 5 and 40℃ and the activity was dependent on pyridoxal 5'-phosphate. K_m and V_max of recombinant GAD were 6.2 ± 0.06 mM and 0.34 ± 0.002 mM/min, respectively. L. brevis B737 can be used as a starter for fermented foods with high GABA contents.

• 대한불안의학회지
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• 제13권2호
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• pp.100-107
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• 2017

Objective : The purpose of this study was to investigate strategies used to cope with stress and the cognitive characteristics of somatic symptom perception in patients with generalized anxiety disorder(GAD). Methods : A total of 55 patients meeting DSM-5 criteria for GAD and 55 normal controls were recruited for participation in this study. We evaluated subjects using The Way of Stress Coping Questionnaire (SCQ), Somato-Sensory Amplification Scale (SSAS), Symptom Interpretation Questionnaire (SIQ), and the Generalized Anxiety Disorder for 7 Item (GAD-7). We analyzed data using an independent t-test and Pearson's correlation analysis. Results : In terms of SCQ, GAD patients presented significantly lower scores on seeking social support and higher scores on wishful thinking than normal controls. GAD patients had significantly greater amplification of physical sensation in SSAS and higher scores in physical, psychological interpretation subsets of SIQ than normal controls. GAD-7 scores were positively correlated with physical interpretation scores on SIQ. Conclusion : Results reveal that patients with GAD have insufficient coping strategies for stress, greater amplification of body sensations, and tendency towards a physical, psychological interpretation of somatic symptoms.

• 핵의학기술
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• 제21권2호
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• pp.28-30
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• 2017

본 연구를 통해 항 GAD 항체검사에서 뇌척수액에 대한 정상치 범위를 추정하였다. 본원에서 211명을 대상으로 혈청과 뇌척수액에 대한 항 GAD 항체검사를 함께 의뢰받은 환자 데이터를 분석해 본 결과 혈청은 1.0 U/mL 이상의 값을 가진 환자가 약 5%를 차지하였고. 뇌척수액의 경우에는 1.0 U/mL이상의 값을 가진 환자는 약 98% 이상을 차지하였다. 혈청의 참고치 (1.0 U/mL 이상 양성)를 뇌척수액에 적용했을 경우에는 대부분의 환자들이 강직증후군 및 간질을 진단 받게 된다. 이에 혈청과 뇌척수액의 참고치를 병용해서 사용하는 것은 부적합하다고 판단하였고 뇌척수액에 대한 참고치를 본원에서 자체적으로 설정하고자 하였다. 실험 대상으로는 정상인의 뇌척수액을 채취하는 데는 어려움이 있어 이와 가장 유사한 생리식염수를 음성대조군으로 선정하였다. 총 70개의 생리식염수를 검사한 결과 Mean값이 1.97 U/mL, SD 값이 0.72 U/mL가 나왔다. 이 값들에 Hoffmann법으로 Mean값에서 ${\pm}2SD$를 정상범위로 하여 0.54 U/mL ~ 3.40 U/mL로 Expected reference range를 설정할 수 있었다. 본 실험에서 아쉬운 점은 참고치 설정에 있어 정상인의 뇌척수액으로 검사가 이루어지지 못한 부분이다. 앞으로 정상인의 뇌척수액으로 참고치 설정을 할 수 있다면 보다 더 정확하게 임상적으로 적용할 수 있을 것이라 생각 된다.

• 한국양식학회지
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• 제3권1호
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• pp.65-77
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• 1990

피조개의 성장 및 적정 양식장의 개발에 관한 기초자료를 얻기 위하여 거제군 사곡리 앞바다에서 천연채묘한 치패를 사용하여 가덕도 및 남해의 피조개 양성장에서 성장비교 실험한 결과는 다음과 같다. 실험개시시의 각장 1.38$\pm$0.32cm와 전중 3.39$\pm$0.30g의 피조개 치패를 17개월간 양성한 결과, 각각의 성장은 가덕도 6.41$\pm$0.30cm, 67.58$\pm$9.63g, 남해 7.20$\pm$0.30cm, 97.33$\pm$11.45g으로 자라나, 남해가 가덕도에 비하여 피조개의 성장이 좋았다. 두 양성장의 피조개 비만도는 가덕도에서 8$\~$9월, 남해에서는 3$\~$5월에 최대치를 보였다. 실험기간중 수온 및 비중은 가덕도에서 $4.9\~27.1^{\circ}C$, 1.0147$\~$1.0259, 남해에서는 $4.5\~26.3^{\circ}C$, 1.0191$\~$1.0263으로, 특히 비중에 있어서 가덕도가 남해에 비하여 낮았다. 입도분석에 의한 저질의 왜도는 남해에서 positive skewness를 나타낸 반면, 가덕도에서는 negative skewness를 보여 가덕도 앞바다의 조류는 남해보다 강한 것으로 나타났다. 양성장간의 성장비교 및 환경 분석에 의한 남해 미조리 지선어장이 가덕도 장항 지선어장에 비하여 조건이 좋은 양성장인 것으로 판단된다.