• Toxicological Research
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• 제3권1호
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• pp.9-14
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• 1987

The effects of monosodium glutamate (MSG) on the activities if tyrosine hydroxylase (TH), dopamine ${\beta}$-hydroxylase (DBH), tryptophan hydroxylase (TPH) and monoamine oxidase (MAO) in various regions (cerebral cortex, striatum, midbrain, pons and medulla of nat brain have been determined. It was observed that up to 1mM MSGhad no significant effects on the activities of brain tyrosine hydroxylase, dopamine ${\beta}$-hydroxylase, tryptophan hydroxylase and monoamine oxidase in all regions of rat brain. These results indicated that MSG itself exerted no direct effect on the important enzymes synthsizing and metabolizing the monoaminergic neuronal system.

• Bulletin of the Korean Chemical Society
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• 제25권5호
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• pp.711-714
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• 2004

Peroxynitrite enters erythrocytes through band 3 anion exchanger and oxidizes cytosolic proteins therein. As a protein associated with band 3, carbonic anhydrase II may suffer from peroxynitrite-induced oxidative damages. Esterase activity of carbonic anhydrase II decreased as the concentration of peroxynitrite increased. Neither hydrogen peroxide nor hypochlorite affected the enzyme activity. Inactivation of the enzyme was in parallel with the release of zinc ion, which is a component of the enzyme's active site. SDS-PAGE of peroxynitrite-treated samples showed no indication of fragmentation but non-denaturing PAGE exhibited new bands with lower positive charges. Western analysis demonstrated that nitration of tyrosine residues increased with the peroxynitrite concentration but the sites of nitration could not be determined. Instead MALDI-TOF analysis identified tryptophan-245 as a site of nitration. Such modification of tryptophan residues is responsible for the decrease in tryptophan fluorescence. These results demonstrate that peroxynitrite nitrates tyrosine and tryptophan residues of carbonic anhydrase II without causing fragmentation or dimerization. The peroxynitrite-induced inactivation of the enzyme is primarily due to the release of zinc ion in the enzyme's active site.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권1호
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• pp.17-22
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• 2000

The separation of tryptophan enantiomers was carried out with medium-pressure liquid chromatography using BSA (bovine serum albumin)-bonded silica as a chiral stationary phase. The influence of various experimental factors such as pH and ionic strength of mobile phase, separation temperature, and the presence of organic additives on the resolution was studied. In order to expand this system to preparative scale, the loadability of sample and the stability of stationary phase for repeated use were also examined. The separation of tryptophan enantiomers was successful with this system. The data indicated that a higher separation factor (a) was obtained at a higher pH and lower temperature and ionic strength in mobile phase. Addition of organic additives (acetonitrile and 2-propanol) in mobile phase contributed to reduce the retention time of L-tryptophan. About $30\%$ of the separation factor was reduced after 80 days of repeated use.

• Mass Spectrometry Letters
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• 제6권3호
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• pp.80-84
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• 2015

Here, we demonstrate the use of MALDI-TOF as a fast and simple analytical approach to evaluate the DNA-binding capability of various peptides. Specifically, by varying the amino acid sequence of the peptides consisting of lysine (K) and tryptophan (W), we identified peptides with strong DNA-binding capabilities using MALDI-TOF. Mass spectrometric analysis reveals an interesting novel finding that lysine residues show sequence selective preference, which used to be considered as mediator of electrostatic interactions with DNA phosphate backbones. Moreover, tryptophan residues show higher affinity to DNA than lysine residues. Since there are numerous possible combinations to make peptide oligomers, it is valuable to introduce a simple and reliable analytical approach in order to quickly identify DNA-binding peptides.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2006년도 제47회 학술심포지움 및 추계국제학술대회
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• pp.77.1-77.1
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• 2006

• Molecules and Cells
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• 제19권2호
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• pp.219-222
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• 2005

The a-subunit of Escherichia coli tryptophan synthase (${\alpha}TS$), a component of the tryptophan synthase ${\alpha}_2{\beta}_2$ complex, is a monomeric 268-residues protein (Mr = 28,600). ${\alpha}TS$ by itself catalyzes the cleavage of indole-3-glycerol phosphate to glyceraldehyde-3-phosphate and indole, which is converted to tryptophan in tryptophan biosynthesis. Wild-type and P28L/Y173F double mutant ${\alpha}$-subunits were overexpressed in E. coli and crystallized at 298 K by the hanging-drop vapor-diffusion method. X-ray diffraction data were collected to $2.5{\AA}$ resolution from the wild-type crystals and to $1.8{\AA}$ from the crystals of the double mutant, since the latter produced better quality diffraction data. The wild-type crystals belonged to the monoclinic space group C2 ($a=155.64{\AA}$, $b=44.54{\AA}$, $c=71.53{\AA}$ and ${\beta}=96.39^{\circ}$) and the P28L/Y173F crystals to the monoclinic space group $P2_1$ ($a=71.09{\AA}$, b=52.70, $c=71.52{\AA}$ and ${\beta}=91.49^{\circ}$). The asymmetric unit of both structures contained two molecules of ${\alpha}TS$. Crystal volume per protein mass ($V_m$) and solvent content were $2.15{\AA}^3\;Da^{-1}$ and 42.95% for the wild-type and $2.34{\AA}^3\;Da^{-1}$ and 47.52% for the double mutant.

• Journal of Microbiology and Biotechnology
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• 제16권6호
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• pp.993-998
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• 2006

We earlier suggested a hierarchical regulatory network using defined modeling symbols and weights in order to improve the flux balance analysis (FBA) with regulatory events that were represented by if-then rules and Boolean logic. In the present study, the simulation results of the models, which were developed and improved from the previou model by incorporating a hierarchical regulatory network into the FBA, were compared with the experimental outcome of an aerobic batch growth of E. coli on glucose and tryptophan. From the experimental result, a diauxic growth curve was observed, reflecting growth resumption, when tryptophan was used as an alternativee after the supply of glucose was exhausted. The model parameters, the initial concentration of substrates (0.92 mM glucose and 1 mM tryptophan), cell density (0.0086 g biomass/1), the maximal uptake rates of substrates (5.4 mmol glucose/g DCW h and 1.32 mmol tryptophan/g DCW h), and lag time (0.32 h) were derived from the experimental data for more accurate prediction. The simulation results agreed with the experimental outcome of the temporal profiles of cell density and glucose, and tryptophan concentrations.

• 한국미생물·생명공학회지
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• 제34권2호
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• pp.121-128
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• 2006

Bacillus alcalophilus AX2000으로부터 xylanase를 정제한 후 효소의 활성부위를 조사하기 위하여 여러 가지 화학수식제를 사용하여 효소활성의 저해도를 측정하였다. 여러 가지 화학 수식제 중에서 carbodiimide와 N-bromosuccinimide가 효소 활성을 완전히 저해시켜 glutamic acid또는 aspartic acid 잔기와 tryptophan 잔기가 효소의 활성부위에 관여하리라 추측되었다. 각각의 경우에 효소 실활은 수식제의 첨가농도에 따라 pseudo first-order kinetics 양식을 보여주었으며, carbodiimide와 N-bromosuccinimide는 각각 비경쟁적 저해와 경쟁적 저해방식을 나타내었다. 기질첨가에 의한 효소활성 보호실험을 통하여 tryptophan 잔기가 기질결합부위라 판단 되었다. 효소 실활속도의 분석에 의해 효소활성에는 2개의 glutamic acid 또는 aspartic acid 잔기와 1개의 tryptophan 잔기가 관여하는 것으로 나타났다.

• 한국미생물·생명공학회지
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• 제14권5호
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• pp.427-432
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• 1986

The regulation of three ammonia assimilatory enzymes, GDH (glutamate dehydrogenase), GS (glutamine synthetase) and GOGAT (glutamate synthase), has been examined in C. glutamicum. Three kinds of arginine auxotrophs blocked in each step of arginine biosynthetic pathway from glutamate were selected as arg 5, arg 6, arg 8. Histidine and tryptophan auxotrophs were also selected because histidine and tryptophan repressed GS biosynthesis in E. coli. These strains were cultured on the media containing nitrogen-excess and limited conditions, to compare the specific activities of ${\alpha}$-ketoglutarate dehydrogenase(${\alpha}-KGDH$), GDH, GS, GOGAT from the cell-free extracts. These results showed that enzyme levels of ${\alpha}-KGDH$ and GDH from 3 kinds of arginine auxotrophs, histidine and tryptophan auxotrophs in nitrogen-excess condition and those of GS and GOGAT in nitrogen limited condition were increased compared with opposite condition. The tryptophan and histidine auxotrophs showed higher level of glutamate and glutamine than parental strains and other mutants. it is assumed that the higher levels of ${\alpha-KGDH}$ and GDH from mutants in nitrogen-excess condition promoted the accumulation of glutamate and glutamine in fermentation broth. The inhibition of GS activities by ADP suggested that GS is regulated by energy charge in C. glutamicum. The results with histidine, tryptophan, glycine, alanine, serine and GMP implied that a system of feedback inhibition were effective. The GDH, GS and GOGAT biosynthesis in culture broth was markedly repressed by the nature and kinds of available nitrogen sources such as tryptophan, proline, glycine, alanine, serine and tyrosine.

• Applied Biological Chemistry
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• 제19권2호
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• pp.65-69
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• 1976

갈대(Phragmites communis Trin) 유아(幼芽)를 $27^{\circ}C$의 암소에서 110시간 동안 사경(砂耕) 배양(培養)하였다. 유아(幼芽)가 자라는 동안 일정(一定)한 시간마다 시료(試料)를 채취하여 methanol로 처리(處理)하여 그 추출물(抽出物)에 대하여 thin layer chromatography를 실시(實施)하였다. 그 결과 배양(培養)하지 않은 유아(幼芽)에서는 indole화합물(化合物) 중에서 serotonin, tryptophan, tryptamine이 다량(多量)으로 나타났으나, 배양(培養) 과정(過程) 중에 이들 화합물(化合物)의 감소(減少)와 더불어 다른 여러 가지 indoleamine류(類)의 생합성(生合成)이 현저(顯著)하였다. 즉 배양(培養) 과정(過程) 중에 tryptophan의 양(量)에는 큰 변화(變化)가 없으나, serotonin 및 tryptamine이 감소하면서, 이들의 methyl화(化) 유도체들이 증가(增加) 하였다. 그 증감(增感)의 상호관계(相互關係)를 관찰하여 본 결과, bufotenine은 tryptophan에서 tryptamine을 거쳐 serotonin, N-methylserotonin의 경로(經路)로 합성(合成)될 것임을 시사하였다.