• 대한화학회지
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• 제31권4호
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• pp.352-358
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• 1987

$25^{\circ}C$의 1 : 4 dioxane-물의 혼합용액속에서 파라-치환된 phenyl N-(p-chlorobenzoyl)chloroformimidate (I) 유도체들의 가수분해 반응속도 상수를 측정하고 반응속도식, 치환기 효과$(\rho\>{\rho}^+)$, 생성물 분석 및 분자궤도 함수의 계산 결과로부터 pH3.0 이하에서는 azocarbonium 이온(II)이 생성되는 $S_N1$반응 메카니즘으로 무촉매 반응이 일어나며, pH 4.0이상에서는 전이상태(III)를 지나는 $S_N2$반응 메카니즘을 통하여 염기 촉매반응이 일어남을 제안 할 수 있었다. 4가지 peri planar형태 이성질체들의 상대적인 안정도는 각각 (E-ap) > (Z-ap) > (E-sp) > 및 (Z-ap)이었고, (E-ap)형태의 가장 안정한 입체구조는 benzimidochloroformyl group면에 대하여 Y-치환 phenyl group이 수직$(90^{\circ})$을 이루었으며 (I)의 활성화된 azomethine탄소 원자에 대하여 물분자는 시그마 공격에 의하여 친핵성 반응이 일어난다.

• Journal of Microbiology and Biotechnology
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• 제16권3호
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• pp.408-413
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• 2006

A strain, P821, with phospholipase D activity was isolated from soil and identified as a Streptomyces species. The phospholipase D enzyme was purified from a culture broth of the isolated strain using ammonium sulfate precipitation and DEAE-Sepharose, phenyl-Sepharose, and Superose 12 HR column chromatographies. The purified enzyme exhibited an optimum temperature and pH of $55^{\circ}C$ and 6.0, respectively, in the hydrolysis of phosphatidylcholine and remained stable up to $60^{\circ}C$ within a pH range of 3.5-8.0. The enzyme also catalyzed a transphosphatidylation reaction to produce phosphatidylserine with phosphatidylcholine and serine substrates. The optimum conditions for the transphosphatidylation were $30^{\circ}C$ and pH 5.0, indicating quite different optimum conditions for the hydrolysis and transphosphatidylation reactions. The gene encoding the enzyme was cloned by Southern hybridization and colony hybridization using a DNA probe designed from the conserved regions of other known phospholipase D enzymes. The resulting amino acid sequence was most similar to that of the PLD enzyme from Streptomyces halstedii (89.5%). Therefore, the enzyme was confirmed to be a phospholipase D with potential use in the production of phosphatidylserine.

• 한국식품과학회지
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• 제22권5호
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• pp.582-589
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• 1990

본 고에서는 Equivalent time과 Equivalent temperature를 활용하여 Kinetic parameters를 결정하는 새로운 방법을 제안하였다. 본 방법의 타당성을 두 가지의 kinetic data 즉, 계산치와 실험치를 이용하여 예시하였다. 계산치는 그 Kinetics가 잘 알려진 세 가지 화학반응에 대해 임의의 등온가열조건을 적용하여 계산하였고 실험치는 2% 설탕용액을 사용하여 0.0005N 염산용액을 사용하여 가수분해가 일어나는 정도를 효소반응을 이용하여 측정하였다. 본 방법에 의해 결정된 활성화 에너지와 Frequency factor는 각각 $104.74{\pm}1.87KJ/mol$과 $5.26{\times}10^{14)hr^{-1}$이었으며 이들 값은 보고된 결과와 잘 일치되었다.

• Bulletin of the Korean Chemical Society
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• 제25권10호
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• pp.1499-1502
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• 2004

An amperometric biosensor based on carbon paste electrodes (CPEs) for the determination of urea was constructed by enzyme (urease/GL-DH)-modified method. Urea was hydrolyzed to ${NH_4}^+$ by catalyzing urease onto the enzyme-modified electrode surface in sample solution. In the presence of ${\alpha}$-ketoglutarate and reduced nicotinamide adenine dinucleotide(NADH), a liberated ${NH_4}^+$ produce to L-glutamate and $NAD^+$ by Lglutamate dehydrogenase (GL-DH). After the chemical reaction was proceeded, the electrochemical reaction was occurred that an excess of the NADH was oxidized to $NAD^+$. The oxidation current of NADH was monitored at +1.10 volt vs. Ag/AgCl. An optimum conditions of biosensor were investigated: The optimum pH range for catalyzed hydrolysis reaction of urea was pH 7.0-7.4. The linear response range and detection limit were $2.0\;{\times}\;10^{-5}{\sim}2.0\;{\times}\;10^{-4}M\;and\;5.0\;{\times}\;10^{-6}M$, respectively. Another physiological species did not interfere, except L-ascorbic acid.

• 공업화학
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• 제25권4호
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• pp.392-395
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• 2014

생리학적 활성을 가진 (2S,3S,4S)-3,4-다이하이드록시 글루타믹산(DHGA)을 값이 저렴하고 수급이 용이한 D-serine 유도체로부터 효율적으로 합성하였다. D-serine 유도체로부터 얻어진 ${\gamma}$-아미노-${\alpha},{\beta}$-불포화성(Z)-에스터의 아민에 다이페닐메틸렌기를 도입, 이를 이용한 입체선택적 이 중 알코올화 반응을 통해 2,3 위치에 두 개의 하이드록시기를 10 : 1 이상의 높은 선택성과 86%의 높은 수율로 도입하여 중간체 5a를 효율적으로 합성하였고, 이 중간체의 간단한 산화 및 가수분해 반응을 통해 (2S,3S,4S)-3,4-DHGA 합성에 성공하였다. 이는 11단계에 총 30%의 수율과 입체선택적인 결과로 현재까지 보고된 (2S,3S,4S)-3,4-DHGA의 합성법 중에 가장 효율적이다. 이 결과는 $OsO_4$을 이용한 입체선택적 이중 알콜화 반응이 아미노 다이올을 포함하는 다양한 생리활성 물질의 효율적인 합성에 적용할 수 있음을 뒷받침한다.

• Journal of Microbiology and Biotechnology
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• 제30권3호
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• pp.391-397
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• 2020

In this study, we used a novel α-L-arabinopyranosidase (AbpBs) obtained from ginsenoside-converting Blastococcus saxobsidens that was cloned and expressed in Escherichia coli BL21 (DE3), and then applied it in the biotransformation of ginsenoside Rb₂ into Rd. The gene, termed AbpBs, consisting of 2,406 nucleotides (801 amino acid residues), and with a predicted translated protein molecular mass of 86.4 kDa, was cloned into a pGEX4T-1 vector. A BLAST search using the AbpBs amino acid sequence revealed significant homology with a family 2 glycoside hydrolase (GH2). The over-expressed recombinant AbpBs in Escherichia coli BL21 (DE3) catalyzed the hydrolysis of the arabinopyranose moiety attached to the C-20 position of ginsenoside Rb₂ under optimal conditions (pH 7.0 and 40℃). Kinetic parameters for α-L-arabinopyranosidase showed apparent K_m and V_max values of 0.078 ± 0.0002 μM and 1.4 ± 0.1 μmol/min/mg of protein against p-nitrophenyl-α-L-arabinopyranoside. Using a purified AbpBs (1 ㎍/ml), 0.1% of ginsenoside Rb₂ was completely converted to ginsenoside Rd within 1 h. The recombinant AbpBs could be useful for high-yield, rapid, and low-cost preparation of ginsenoside Rd from Rb₂.

• Journal of Microbiology and Biotechnology
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• 제14권6호
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• pp.1134-1141
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• 2004

A bacterial strain capable of hydrolyzing sulfate ester bonds in p-nitrophenyl sulfate and agar was isolated from the Southeast coast of Korea. The isolated strain (AS6330) is aerobic, Gram-negative, rod-shaped, and motile. Octadecanoic acid was the major cellular fatty acid in the isolate. An almost complete 16S rDNA sequence of the isolate was determined and the sequence similarity of the 16S rDNA with those of known Sphingomonas spp. was found to be at most $96.4\%$, implying that the isolate was a new Sphingomonas species. The organism was grown optimally at NaCl concentration of $1.5-3.5\%$. Optimum culture conditions were determined to be $30^{\circ}C$ and pH 7.0 for 48 h fermentation using a laboratory fermentor under constant culture conditions. Partially purified arylsulfatase through Q-Sepharose and phenyl­Sepharose chromatographies catalyzed hydrolysis of sulfate ester bonds in agar, and $97\%$ of sulfates in agar were removed after 4 h reaction at $45^{\circ}C$ and pH 7.0. The arylsulfatase from the isolated bacterium might be useful for the removal of sulfate groups in agar.

• Journal of Microbiology and Biotechnology
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• 제21권8호
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• pp.830-837
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• 2011

A genomic DNA fragment encoding a putative maltooligosyltrehalose trehalohydrolase (NfMTH) for trehalose biosynthesis was cloned by the degenerate primer- PCR from cyanobacterium Nostoc flagelliforme. The ORF of NfMTH is 1,848 bp in length and encodes 615 amino acid residues, constituting a 70 kDa protein. The deduced amino acid sequence of NfMTH contains 4 regions highly conserved for MTHs. By expression of NfMTH in E. coli, the function of this protein was demonstrated, where the recombinant protein catalyzed the hydrolysis of maltooligosyl trehalose to trehalose. The expressions of MTH and maltooligosyltrehalose synthase in the filaments of N. flagelliforme were upregulated significantly under dehydration stress, NaCl stress, and high temperature-drought stress. The accumulations of both trehalose and sucrose in the filaments of N. flagelliforme were also improved significantly under the above stresses. Furthermore, trehalose accumulated in smaller quantities than sucrose did when under NaCl stress, but accumulated in higher quantities than sucrose did when under temperature-drought stress, indicating that both trehalose and sucrose were involved in N. flagelliforme adapted to stresses and different strategies conducted in response to various stress conditions.

• Journal of Microbiology and Biotechnology
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• 제16권2호
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• pp.272-279
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• 2006

The gene encoding Thermus sp. T351 alkaline phosphatase (T351 APase) was cloned and sequenced. The gene consisted of 1,503 bp coding for a protein with 500 amino acid residues including a signal peptide. The deduced amino acid sequence of T351 APase showed relatively low similarity to other Thermus APases. The T351 APase gene was expressed under the control of the T7lac promoter on the expression vector pET-22b(+) in Escherichia coli BL21 (DE3). The expressed enzyme was purified by heat treatment, and $UNO^{TM}$ Q and $HiTrap^{TM}$ Heparin HP column chromatographies. The purified enzyme exhibited high activity at extremely alkaline pHs, reaching a maximum at pH 12.0. The optimum temperature of the enzyme was $80^{\circ}C$, and the half-life at $85^{\circ}C$ was approximately 103 min. The enzyme activity was found to be dependent on metal ions: the addition of $Mg^{2+}$ and $CO^{2+}$ increased the activity, whereas EDTA inhibited it. With p-nitrophenyl phosphate as the substrate, T351 APase had a Michaelis constant ($K_{m}$) of $3.9{\times}10^{-5}M$. The enzyme catalyzed the hydrolysis of a wide variety of phosphorylated compounds.

• 임산에너지
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• 제21권2호
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• pp.25-33
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• 2002

도서관이나 문서보관소에서 책이나 서류를 보존하는데 있어 중요한 문제점 중의 하나는 종이의 열화이다. 그러므로 이러한 문제를 해결하기 위해서는 종이의 열화의 주요 원인을 규명할 필요가 있다. 몇몇의 학자들이 이에 대한 연구를 수행한 결과 종이 열화의 주요 원인으로서 종이 섬유내 셀룰로오스의 산 촉매 가수분해로 밝혀졌다. 일반적으로 종이제조시 첨가되는 첨가제로 인해 산성지의 노화율이 중성지의 노화율에 비해 더 높다. 따라서 종이내 존재하는 산을 제거해 줄 필요가 있다. 종이의 탈산화 처리는 열화속도를 감소시켜 산성지의 수명을 3, 4배정도 늘릴 수 있다고 한다. 최근에는 기존의 탈산화 처리방법과는달리 대량의 책과 서류의 탈산화를 위한 효과적인 탈산화 방법의 필요성이 인식되어져 왔다. 따라서 본 논문에서는 종이 제조시 사용되는 첨가제가 노화에 미치는 영향을 알아보고,효과적인 종이의 노화 방지를 위해 산성지에 가스상 에탄올아민류(모노에탄올아민, 디에탄올아민, 트리에탄올아민)를 사용하여 탈산처리를 시도하여 보았다. 첨가제가 종이의 노화에미치는 영향을 실험한 결과, 종이의 노화율이 알럼＋로진＞알럼＞AKD＞ 무처리 순으로 나타났다. 또한 가스상 에탄올아민류 탈산처리 실험결과, 탈산처리율이 모노에탄올아민＞디에탄올아민＞트리에탄올아민 순으로 나타났다. 그러나 처리 후 종이에 약간의 백색도와 내절도의 감소가 나타났다. 이러한 문제를 해결하기 위해 가스상 에탄올아민류들을 조합 처리하여 탈산처리를 시도해 본 결과, 백색도와 내절도의 감소없이 효과적인 탈산처리 효과를 얻을 수 있었다.ird limit" is characterized by the production of reduction of $H_2O$$_2$, which is reduced by wall effect. Strain rate substantially affects ignition temperature because key reaction rates of $H_2O$$_2$ are comparably with its transport rate, while the mixture temperature and the hydrogen composition do not significantly affect ignition temperature.e.성 면역 결핍증 환자가 직업을 가지면 안된다는 사람보다 된다는 사람이(P<0.001), 각각 지식이 높았다. 결론 : 지식이 높을수록 성병에 걸릴 가능성이 낮고, 태도에서도 긍정적인 결과를 보였다. 그러므로 그러므로 가능하면 중고교 시절에 이 질환에 대한 정규교육 프로그램을 만들어 학생들을 효과적으로 가르치는 것이 필요하다.와 활동성 정도(r=0.378, P<.05), 평균 통증정도와 활동성 정도가(r=.330, P<.05)가 유의한 정적상관관계가 나타난 반면, 여성에서는 활동성 정도와 통증의 중증도는 유의한 관계가 없는 것으로 나타났다. 남성은 관계를 제외한 모든 항목의 통증으로 인한 지장정도와 활동성 정도가 유의한 정적상관관계가 나타난 반만 여성에서는 보행 능력, 통상적인