• BMB Reports
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• v.42 no.1
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• pp.47-52
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• 2009

Alanine racemase catalyzes the interconversion of L-alanine and D-alanine and plays a crucial role in spore germination and cell wall biosynthesis. In this study, alanine racemase produced by Bacillus anthracis was expressed and purified as a monomer in Escherichia coli and the importance of lysine 41 in the cofactor binding octapeptide and tyrosine 270 in catalysis was evaluated. The native enzyme exhibited an apparent $K_m$ of 3 mM for L-alanine, and a $V_{max}$ of $295\;{\mu}moles/min/mg$, with the optimum activity occurring at $37^{\circ}C$ and a pH of 8-9. The activity observed in the absence of exogenous pyridoxal 5'-phosphate suggested that the cofactor is bound to the enzyme. Additionally, the UV-visible absorption spectra indicated that the activity was pH independece, of VV-visible absorption spectra suggests that the bound PLP exists as a protonated Schiff's base. Furthermore, the loss of activity observed in the apoenzyme suggested that bound PLP is required for catalysis. Finally, the enzyme followed non-competitive and mixed inhibition kinetics for hydroxylamine and propionate with a $K_i$of $160\;{\mu}M$ and 30 mM, respectively.

• IE interfaces
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• v.7 no.2
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• pp.61-69
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• 1994

기업의 제품책임(PL)은 사전책임(PLP)과 사후책임(PLD)으로 크게 나눌 수 있다. 제품책임에 대한 선진국 기업의 조직구조를 분석해보면 안전책임, 품질보증책임, 제품정보 공개책임, 피해보상 및 A/S, 제품책임 보험, 제품책임소송 등을 담당하여 처리하고 있음을 알 수 있다. 본 논문은 ISO-9000에서 요구하는 사항중에서 기업의 제품책임에 관한 내용을 살펴보고, 이에 대한 기업의 대응방향을 고찰하고자 한다.

• Proceedings of the Korean Institute of Industrial Safety Conference
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• 2003.05a
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• pp.43-47
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• 2003

현대 과학기술의 발달과 더불어 생산되는 제품들은 점차 고도화·복잡화되고, 제품의 결함 또는 제품으로 인한 사고 원인을 소비자 스스로 밝혀내는 것이 거의 불가능하게 되었다. 따라서 제조물의 안전에 대한 우선적인 입증책임을 기업에게 요구하는 제조물책임법이 선진국에서 시행되었고, 최근 소비자의 권익 보호제도 확충으로 제조물에 대한 책임이 엄격화, 광범위화 되었으며, 유럽연합과 일본의 제조물책임법을 근간으로 제정한 국내의 제조물책임법이 2002년 7월 시행되었다.(중략)

• Proceedings of the Acoustical Society of Korea Conference
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• 1995.06a
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• pp.184-187
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• 1995

This paper proposes an algorithm for machine recognition of phonemes in continuous speech. The proposed algorithm is static strategy neural network. The algorithm uses, at the stage of training neuron, features such as PARCOR coefficient and auditory-like perceptual liner prediction . These features are extracted from speech samples selected by a sliding 25.6msec windows with s sliding gap being 3 msec long, then interleaved and summed up to 7 sets of parmeters covering 171 msec worth of speech for use of neural inputs. Perfomances are compared when either PARCOR or auditory-like PLP is included in the feture set.

• Journal of Microbiology and Biotechnology
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• v.18 no.1
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• pp.55-58
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• 2008

Alanine racemase, a bacterial enzyme belonging to the fold-type III group of pyridoxal 5'-phosphate (PLP)-dependent enzymes, has been shown to catalyze the interconversion between L- and D-alanine. The alanine racemase from the pathogenic bacterium Enterococcus faecalis v583 has been overexpressed in E. coli and was shown to crystallize an enzyme at 295 K, using polyethylene glycol (PEG) 8000 as a precipitant. X-ray diffraction data to $2.5{\AA}$ has been collected using synchrotron radiation. The crystal is a member of the orthorhombic space group, $C222_1$ with unit cell parameter of a=94.634, b=156.516, $c=147.878{\AA},\;and\;{\alpha}={\beta}={\gamma}=90{\AA}$. Two or three monomers are likely to be present in the asymmetric unit, with a corresponding $V_m\; of\;3.38{\AA}^3\;Da^{-1}\;and\;2.26{\AA}^3\;Da^{-1}$ and a solvent content of 63.7% and 45.5%, respectively.

• Journal of Applied Biological Chemistry
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• v.54 no.4
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• pp.231-237
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• 2011

A gene encoding a putative alanine racemase in Xanthomonas. oryzae pv. oryzae was cloned, expressed and characterized. Expression of the cloned gene was performed in Escherichia coli BL21(DE3)pLys using a pET-21(a) vector harbouring $6{\times}histidine$ tag. Purification of the recombinant alanine racemase by affinity chromatography resulted in major one band by sodium dodecyl sulfate polyacryl amide gel electrophoresis analysis, showing about 45 kDa of molecular weight. The alanine racemase gene, cloned in this experiment, appears to be constitutively expressed in X. oryzae, as analyzed by reverse transcriptase polymerase chain reaction. The enzyme was the most active toward L-alanine and secondly D-alanine, showing a racemic reaction, thus the enzyme is considered as an alanine racemase. The enzyme was considerably activated by addition of pyridoxal-5-phosphate (PLP), showing that 75% increase in activity was observed at 0.3 mM, compared with control. D-Cysteine as well as L-cysteine significantly inhibited the enzyme activity. The inhibitions by cysteines were more prominent in the absence of PLP, showing 9 and 5% of control activity at 2 mM of addition, respectively. The enzyme was the most active at pH 8.0 and more stable at alkaline pHs than acidic pH condition.

• Journal of Microbiology and Biotechnology
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• v.25 no.7
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• pp.1108-1113
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• 2015

Cadaverine (1,5-diaminopentane) is an important industrial chemical with a wide range of applications. Although there have been many efforts to produce cadaverine through fermentation, there are not many reports of the direct cadaverine production from lysine using biotransformation. Whole-cell reactions were examined using a recombinant Escherichia coli strain overexpressing the E. coli MG1655 cadA gene, and various parameters were investigated for the whole-cell bioconversion of lysine to cadaverine. A high concentration of lysine resulted in the synthesis of pyridoxal-5'-phosphate (PLP) and it was found to be a critical control factor for the biotransformation of lysine to cadaverine. When 0.025 mM PLP and 1.75 M lysine in 500 mM sodium acetate buffer (pH6) were used, consumption of 91% lysine and conversion of about 80% lysine to cadaverine were successfully achieved.

• Journal of the Korean Institute of Intelligent Systems
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• v.15 no.6
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• pp.655-660
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• 2005

This paper studied the feature parameters less affected by the emotional variation for the development of the robust speech recognition technologies. For this purpose, the effect of emotional variation on the speech recognition system and robust feature parameters of speech recognition system were studied using speech database containing various emotions. In this study, LPC cepstral coefficient, met-cepstral coefficient, root-cepstral coefficient, PLP coefficient, RASTA met-cepstral coefficient were used as a feature parameters. And CMS and SBR method were used as a signal bias removal techniques. Experimental results showed that the HMM based speaker independent word recognizer using RASTA met-cepstral coefficient :md its derivatives and CMS as a signal bias removal showed the best performance of $7.05\%$ word error rate. This corresponds to about a $52\%$ word error reduction as compare to the performance of baseline system using met - cepstral coefficient.

• BMB Reports
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• v.32 no.5
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• pp.480-485
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• 1999

Tyrosine phenol-lyase of thermophilic Symbiobacterium sp. SC-1, which is obligately and symbiotically dependent on thermophilic Bacillus sp. SK-1, was purified and characterized. The enzyme is composed of four identical subunits and contains approximately 1 mol of pyridoxal 5'-phosphate (PLP) per mol subunit as a cofactor. The enzyme showed absorption maxima at 330 and 420 nm, and lost this absorption profile by treatment with phenylhydrazine. The apparent dissociation constsnt, $K'_D$, for PLP was determined with the apoenzyme to be about $1.2\;{\mu}M$. The isoelectric point was 4.9. The optimal temperature and pH for the $\alpha,\beta$-elimination of L-tyrosine were found to be $80^{\circ}C$ and pH 8.0, respectively. The substrate specificity of the enzyme was very broad: L-amino acids including L-tyrosine, 3,4-dihydroxyphenyl-L-alanine (L-DOPA), L-cysteine, L-serine, S-methyl-L-cysteine, $\beta$-chloro-L-alanine, and S-(o-nitrophenyl)-L-cysteine all served as substrates. D-Tyrosine and D-serine were also decomposed into pyruvic acid and ammonia at rates of 7% and 31% relative to their corresponding L-enantiomers, respectively. D-Alanine, which was inert as a substrate in a, $\beta$-elimination, was the only D-amino acid racemized by the enzyme. The $K_m$ values for L-tyrosine, L-DOPA, S-(o-nitrophenyl)-L-cysteine, $\beta$-chloro-L-alanine, and S-methyl-L-cysteine were 0.19, 9.9, 0.36, 12, and 5.5 mM, respectively.

• Clinical and Experimental Pediatrics
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• v.55 no.10
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• pp.397-402
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• 2012

Pelizaeus-Merzbacher disease (PMD) is a rare, X-linked recessive disorder characterized by dysmyelination in the central nervous system. PMD results from deletion, mutation, or duplication of the proteolipid protein gene (PLP1) located at Xq22, leading to the failure of axon myelination by oligodendrocytes in the central nervous system. PMD may be suspected when there are clinical manifestations such as nystagmus, developmental delays, and spasticity, and genetic analysis can confirm the diagnosis. Further diagnostic manifestations of the disease include a lack of myelination on brain magnetic resonance (MR) imaging and aberrant N-acetyl aspartate (NAA) and choline concentrations that reflect axonal and myelination abnormalities on phroton MR spectroscopy. We report 5 cases of PMD (in 1 girl and 4 boys). PLP1 duplication was detected in 2 patients. Brain MR analyses and MR spectroscopy were performed for all the patients. The brain MR images showed white matter abnormalities typical of PMD, and the MR spectroscopic images showed diverse patterns of NAA, creatinine, and choline concentrations. We propose that MR spectroscopic analysis of metabolic alterations can aid the PMD diagnosis and can contribute to a better understanding of the pathogenesis of the disease.