• 약학회지
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• 제38권6호
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• pp.791-794
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• 1994

Sixteen kinds of naturally occurring phenolic compounds including 5 stilbenes, 7 flavonoids and 4 anthraquinones were examined in the inhibitory activity against rat liver AADC(aromatic L-amino acid decarboxylase) in vitro, using 5-hydroxytryptophan as a substrate. Three hydroxystilbenes, resveratrol 1, rhapontigenin 3 and piceatanol 5, which were known to be monoamine oxidase A inhibitors, exhibited a significant inhibition against AADC($IC_{50}$=20, 8 and $5\;{\mu}M$, respectively). By the comparison of the activity of each phenolic compound, it was suggested that the 3',4'-dihydroxyphenyl group of stilbenes or flavones was the best pharmacophore for the AADC inhibitory activity.

• 생약학회지
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• 제34권1호통권132호
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• pp.55-59
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• 2003

The effects of liriodenine, an aporphine isoquinoline alkaloid, on dopamine content in PCl2 cells were investigated. Treatment of PC12 cells with liriodenine decreased dopamine content in a dose-dependent manner (33.6% inhibition at $10\;{\mu}M$ for 12 h). The $IC_{50}$ in value of liriodenine was $8.4\;{\mu}M$. Dopamine content decreased at 3 h and reached a minimal level at 12 h after the exposure to liriodenine. Under these conditions, the activities of tyrosine hydroxylase and aromatic L-amino acid decarboxylase were also inhibited at $10\;{\mu}M$ of liriodenine by 10.1% and 20.2% relative to control, respectively. In addition, liriodenine inhibited the increase in dopamine content induced by L-DOPA Treatments $(50-100\;{\mu}M)$ in PC12 cells. These results suggest that liriodenine inhibited dopamine biosynthesis and L-DOPA-induced increase in dopamine content by reducing the activities of tyrosine hydroxylase and aromatic L- amino acid decarboxylase in PC12 cells.

• BMB Reports
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• 제38권5호
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• pp.595-601
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• 2005

In this study, we have isolated a rice (Oryza sativa L.) glutamate decarboxylase (RicGAD) clone from a root cDNA library, using a partial Arabidopsis thaliana GAD gene as a probe. The rice root cDNA library was constructed with mRNA, which had been derived from the roots of rice seedlings subjected to phosphorus deprivation. Nucleotide sequence analysis indicated that the RicGAD clone was 1,712 bp long, and harbors a complete open reading frame of 505 amino acids. The 505 amino acid sequence deduced from this RicGAD clone exhibited 67.7% and 61.9% identity with OsGAD1 (AB056060) and OsGAD2 (AB056061) in the database, respectively. The 505 amino acid sequence also exhibited 62.9, 64.1, and 64.2% identity to Arabidopsis GAD (U9937), Nicotiana tabacum GAD (AF020425), and Petunia hybrida GAD (L16797), respectively. The RicGAD was found to possess a highly conserved tryptophan residue, but lacks the lysine cluster at the C-proximal position, as well as other stretches of positively charged residues. The GAD sequence was expressed heterologously using the high copy number plasmid, pVUCH. Our activation analysis revealed that the maximal activation of the RicGAD occurred in the presence of both $Ca^{2+}$ and calmodulin. The GAD-encoded 56~58 kDa protein was identified via Western blot analysis, using an anti-GAD monoclonal antibody. The results of our RT-PCR analyses revealed that RicGAD is expressed predominantly in rice roots obtained from rice seedlings grown under phosphorus deprivation conditions, and in non-germinated brown rice, which is known to have a limited phosphorus bioavailability. These results indicate that RicGAD is a $Ca^{2+}$/calmodulin-dependent enzyme, and that RicGAD is expressed primarily under phosphate deprivation conditions.

• 생약학회지
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• 제32권1호통권124호
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• pp.49-54
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• 2001

The effects of protoberberine compounds on serotonin biosynthesis in P815 cells were investigated. Protoberberine compounds such as berberine, palmatine and coralyne decreased serotonin content dose-dependently, but coptisine did not. The $IC_{50}$ values of berberine, palmatine and coralyne were $3.0\;{\mu}M,\;16.5\;{\mu}M\;and\;14.5\;{\mu}M$, respectively. Protoberberine compounds at concentrations up to $20\;{\mu}M$ were not cytotoxic towards P815 cells. The activity of tryptophan hydroxylase, a ratelimiting enzyme in serotonin biosynthesis, was inhibited by the exposure of berberine, palmatine and coralyne in P815 cells (14.9-19.3% inhibition at $2-15\;{\mu}M$), but that of aromatic L-amino acid decarboxylase was not. These results suggest that the inhibition of tryptophan hydroxylase activity by berberine, palmatine and coralyne might partially contribute to the decrease in serotonin content in P815 cells.

• 한국버섯학회지
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• 제11권4호
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• pp.181-186
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• 2013

${\gamma}$-Aminobutyric acid(GABA) is a four carbon non-protein amino acid that has several well-known physiological functions, such as a postsynaptic inhibitory neurotransmitter in the brain and induction of hypotensive and tranquilizer effects. A lactic acid bacterium was isolated from button mushroom bed, which is showing high GABA productivity by TLC or HPLC analysis. The strain was identified as Lactobacillus hilgardii by analysis of 16S rDNA gene sequence. When the maximum production of GABA by L. hilgardii was investigated with various concentration of monosodium glutamate, the yield of GABA reached to be 53.65 mM at 1% mono sodium glutamate (MSG) in flask cultivation. A Glutamate decarboxylase (GAD) enzyme, which was known to convert MSG to GABA, was purified from a cell-free extract of L. hilgardii and the molecular weights of purified GAD was estimated to 60,000 by SDS-PAGE. The optimum pH and temperature of GAD were at pH4.6 and at $37^{\circ}C$, respectively. The GAD activity was increased by the addition of sulfate ions such as ammonium sulfate, sodium sulfate and magnesium sulfate, indicating that the increase of hydrophobic interaction causes the increase of GAD activity.

• Natural Product Sciences
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• 제10권3호
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• pp.119-123
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• 2004

The effects of BHSH on L-DOPA-induced increase in dopamine content in PC12 cells were investigated. L-DOPA treatment at $20\;or\;50\;{\mu}M$ increased dopamine content after both 24 and 48 h of incubation in PC12 cells. However, the co-treatments of BHSH $(10-50\;{\mu}M)$ with L-DOPA $(20\;or\;50\;{\mu}M)$ significantly inhibited the increase of dopamine content induced by L-DOPA. BHSH treatment at $10-50\;{\mu}M$ significantly inhibited basal aromatic L-amino acid decarboxylase (AADC) activity in a concentration-dependent manner at 15 min, and then AADC activity was rapidly recovered to the control level at about 2 h. These results indicate that the inhibition of AADC activity by BHSH was, in part, contributed to the early-stage decrease of dopamine content induced by LDOPA in PC12 cells. Taken together, it is proposed that the short-term inhibition of dopamine biosynthesis by BHSH was mediated by the regulation of tyrosine hydroxylace (TH).

• Applied Biological Chemistry
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• 제39권2호
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• pp.112-117
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• 1996

ADC는 diamine인 putrescine 생합성의 두가지 경로중에서 식물계에서 특히 중요한 효소이며, ADC 유전자는 E. coli, 귀리, 토마토 genome에서 이미 cloning된 바 있다. 벼 (Oryza sativa L.) 게놈 DNA의 PCR 증폭을 위해서 토마토와 E. coli의 ABC cDNA의 보존된 부분과 일치하는 두개의 degenerate oligonucleotides (17mer)를 인위 합성하였으며, 증폭의 결과 약 1 kbp 크기의 DNA가 관찰되었다. 증폭된 DNA 절편은 1,022bp 염기서열을 포함하고 있는 ORE (open reading frame)으로 확인되었다. 이 PCR product는 POEM-originated T vector에 재조합하였으며 PstI 제한효소로 약 500bp 크기로 절단하여 pGEM-3Zf(+/-) vector에 subcloning하였다. 벼 ADC clone의 염기서열은 귀리와 토마토 ADC cDNA 서열의 같은 부분과 각각 74%와 70%의 동질성을 갖는 것으로 나타났으며, 예상되는 아미노산 서열은 귀리와 토마토 ADC 단백질과 각각 45%와 62%의 동질성이 관찰되었다. 귀리와 E. coli, 토마토와 귀리 그리고 토마토와 E. coli ADC 아미노산 서열에서 각각 34%, 47%, 그리고 38%의 유사성 정도가 보고된 것을 비교하여 볼 때, 벼와 귀리 및 토마토 사이의 유사성 정도는 다른 비교 보다도 월등히 높았다. 벼 유묘기 잎조직에서 추출한 RNA를 이용한 Northern blot 분석에서 ADC는 약 2.5kbp의 전사체로 발현됨이 확인되었다.

• Journal of Microbiology and Biotechnology
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• 제21권10호
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• pp.1049-1052
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• 2011

In this study, we demonstrated the asymmetric synthesis of L-tert-leucine from trimethylpyruvate using branched-chain aminotransferase (BCAT) from Escherichia coli in the presence of L-glutamate as an amino donor. Since BCAT was severely inhibited by 2-ketoglutarate, in order to overcome this here, we developed a BCAT/aspartate aminotransferase (AspAT) and BCAT/AspAT/pyruvate decarboxylase (PDC) coupling reaction. In the BCAT/AspAT/PDC coupling reaction, 89.2 mM L-tert-leucine (ee>99%) was asymmetrically synthesized from 100 mM trimethylpyruvate.

• 생명과학회지
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• 제16권1호
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• pp.44-48
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• 2006

L-alanine의 산업적 생산을 위한 신규의 L-aspartate $\beta-carboxylase$ 유전자를 Enterococcus faecalis에서 검색하고 이를 대장균에 형질전환시켰다. E. faecalis 유래의 ADC유전자 는 1611 bp의 염기서열로 구성되어 있으며 형질전환된 대장균에서 59 KDa의 효소를 생산하며 L-aspartate $\beta-carboxylase$의 촉매활성을 나타내는 것을 확인하였다. 본 연구결과로 기능은 모르고 유전체 서열만 아는 균체에서 신규 효소를 개발하는 방법을 확립하였으며 저가의 aspartate를 이용한 고부가가치 L-alanine을 생산할 수 있는 신규효소를 개발할 수 있었다.

• Asian-Australasian Journal of Animal Sciences
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• 제32권10호
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• pp.1580-1590
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• 2019

Objective: The aims of this study were to select one strain of Lactobacillus plantarum (L. plantarum) for a potential indigenous safe starter culture with low level antibiotic resistant and low biogenic amine production and evaluate its effect on biogenic amines reduction in Moo som. Methods: Three strains of indigenous L. plantarum starter culture (KL101, KL102, and KL103) were selected based on their safety including antibiotic resistance and decarboxylase activity, and fermentation property as compared with a commercial starter culture (L. plantarum TISIR543). Subsequently, the effect of the selected indigenous safe starter culture on biogenic amines formation during Moo som fermentation was studied. Results: KL102 and TISIR 543 were susceptible to penicillin G, tetracycline, chloramphenicol, erythromycin, gentamycin, streptomycin, vancomycin, ciprofloxacin and trimethoprim (MIC90 ranging from 0.25 to $4{\mu}g/mL$). All strains were negative amino acid-decarboxylase for lysis of biogenic amines in screening medium. For fermentation in Moo som broth, a relatively high maximum growth rate of KL102 and TISIR543 resulted in a generation time than in the other strains (p<0.05). These strain counts were constant during the end of fermentation. Similarly, KL102 or TISIR543 addition supported increases of lactic acid bacterial count and total acidity in Moo som fermentation. For biogenic amine reduction, tyramine, putrescine, histamine and spermine contents in Moo som decreased significantly by the addition KL102 during 1 d of fermentation (p<0.05). In final product, histamine, spermine and tryptamine contents in Moo som inoculated with KL102 were lower amount those with TISIR543 (p<0.05). Conclusion: KL102 was a suitable starter culture to reduce the biogenic amine formation in Moo som.