• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.554-556
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• 2003

본 연구에서는 고부가가치 의약단백질인 human growth hormone을 고순도로 얻기 위하여 재조합 대장균을 이용하여 ubiquitin이 융합된 형태로 단백질을 발현시키고, 이를 분해하는 ubiquitin-specific pretense를 발현시켜 이를 분리 ${\cdot}$ 정제하고 효소특성을 살펴보았다. UBP1 enzyme을 재조합 대장균을 이용하여 발현하고 분리 ${\cdot}$ 정제한 결과, 분자량은 약 83.5kDa이었으며, $40^{\circ}C$, pH 8.0에서 최대 효소활성을 보였다.

• Journal of Ginseng Research
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• 제14권1호
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• pp.14-20
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• 1990

In An invertase (EC 3.2.1.26) was extracted from Korean giseng (Panax ginseng C.A. Meyer) with distilled tvater The ginseng invertase was purified about 62.6 folds purified by procedures including ammonium sulfate fractionation , DEAE-cellulofine chromatography and gelfiltrations through Sephadex G-75 and the recovery of enzyme activity was 11.1%. The homogeneity of the purified enzyme was probed by polyacrylamide gel disc electrophoresis. The purifled enzyme was divided into two different subunits by treating with a mixture of SDS and 2-mercautoethanol, and the molecular weight of the large subunit was estimatedtobe 116,000 and that of the small one to be 14,000. The optimal VH and temperature of the enzyme were pH 6 and 45$^{\circ}C$, respectively. The enzyme hydrolyzed specifically the hydrolyzation of the -fructofuranosides such as sucrose, raffinose and inulin. The Km values of the enzyme for sucrose and raffinose were determined to be 0.85 and 0.6 mM, respectively.

• 한국미생물·생명공학회지
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• 제16권3호
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• pp.219-225
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• 1988

Trichoderma viride가 분비하는 섬유소 분해효소를 황산암모늄침전, Sephadex G-100 및 DEAE Sephadex column을 통과시켜서 5개의 섬유소 분해 아이조자임을 분리하였으며 전기영동 방법에 의해서 2개의 celloblohydrolase와 15개의 endoglucanase로 분리하였다. 분리된 cellobiohydrolase는 분자량이 71,000, 최적 pH5.1, 최적온도 5$0^{\circ}C$, pI가 3.81이었으며 주종아미노산은 aspartric acid, glutamic acid. threonine, serine 및 glycine이었다.

• 한국미생물·생명공학회지
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• 제22권1호
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• pp.65-72
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• 1994

Purification and characterization of pectate lyase from Bacillus sp HSA-925. Bacillus sp. HSA-925 isolated from soil produced constitutively an extracellular pectate lyase when cultivated in LB broth. The pectate lyase(EC 4.2.2.2) was purified from the cuylture broth by preciptation with ammonium sulfate, followed by column chromatography on CM-cellulose C-50 and repeated gel filtration on Sephadex G-75G. The enzyme had a molecular weight of 32-33 kDa. The activity was mazimum at pH 9.5 AND 45$\CIRC $C. The enzume activity was stable at 55$\circ $C for 15 min and between pH7-12. The activation energy, Km and V$_{max}$ for the pectate lyase were 5.8779 kcal/mol, 6.33$\times $10$^{-2}$ mol/ml and 2.09$\times $10$^{2}$ $\mu $mol/min respectively. The enzyme was activated by Ca$^{2+}$, Cu$^{2+}$ and inhibited by Li$^{+}$, Hg$^{2+}$, EDTA.

• 한국식품영양학회지
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• 제7권1호
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• pp.16-22
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• 1994

Xylanase from Bacillus sp. GS was purified through acetone precipitation, DEAE-Sephadex A-50 ion exchange chromatography and Sephadex G-100 gel filtration. The optimum reaction temperature of purified xylanase was 50t . Its optimum pH was between pH 6.0 and pH 6.5. This enzyme was stable below 5$0^{\circ}C$ for several hours and stable at between pH 5.5 and pH 8.0. The enzyme activity of xylanase was remarkably increased by Co++ and Cu++ ions. According to the study of hydrolysis mode of this enzyme, it was turned out to be ends type xylanase that can produce xylooligosaccharides, known as bifidogenic factor, from xylan.

• Journal of Microbiology and Biotechnology
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• 제26권2호
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• pp.255-262
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• 2016

The carrageenan-degrading marine bacterium Vibrio sp. strain NJ-2 was isolated from rotten red algae, and κ-carrageenase with high activity was purified from the culture supernatant. The purified enzyme with molecular mass of 33 kDa showed the maximal activity of 937 U/mg at 40℃ and pH 8.0. It maintained 80% of total activity below 40℃ and between pH 6.0 and 10.0. The kinetics experiment showed the K_m and V_max values were 2.54 g/ml and 138.89 mmol/min/mg, respectively. The thin layer chromatography and ESI-MS analysis of hydrolysates indicated that the enzyme can endolytically depolymerize the κ-carrageenan into oligosaccharides with degrees of depolymerization of 2-8. Owing to its high activity, it could be a valuable tool to produce κ-carrageenan oligosaccharides with various biological activities.

• BMB Reports
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• 제45권2호
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• pp.91-95
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• 2012

Glutamate dehydrogenase from axenic bacterial cultures of a new microorganism, called GWE1, isolated from the interior of a sterilization drying oven, was purified by anion-exchange and molecular-exclusion liquid chromatography. The apparent molecular mass of the native enzyme was 250.5 kDa and was shown to be an hexamer with similar subunits of molecular mass 40.5 kDa. For glutamate oxidation, the enzyme showed an optimal pH and temperature of 8.0 and $70^{\circ}C$, respectively. In contrast to other glutamate dehydrogenases isolated from bacteria, the enzyme isolated in this study can use both $NAD^+$ and $NADP^+$ as electron acceptors, displaying more affinity for $NADP^+$ than for $NAD^+$. No activity was detected with NADH or NADPH, 2-oxoglutarate and ammonia. The enzyme was exceptionally thermostable, maintaining more than 70% of activity after incubating at $100^{\circ}C$ for more than five hours suggesting being one of the most thermoestable enzymes reported in the family of dehydrogenases.

• 한국미생물·생명공학회지
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• 제22권6호
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• pp.627-635
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• 1994

The $\beta$-xylosidase from Penicillium sp. FX-102 was purified by 40~80% ammonium sulfate saturation, CM-Cellulose column chromatography, Sephadex G-200 gel filtration, and isoelec- tric focusing. The optimum pH and temperature for the activity of the $\beta$-xylosidase was pH 4.5 and 50$\circ$C, respectively. The enzyme was stable at the pH range of 4.5~5.5, and at 55$\circ$C for 10 min. The molecular weight of the enzyme was estimated to be about 300,000 daltons by Sephadex G-200 gel filtration and 310,000 daltons of monomer by SDS polyacrylamide gel electrophoresis. Isoelectric point of the enzyme was determined to be pH 4.4. The enzyme activity was strongly inhibited by Hg$^{2+}$, Ag$^{2+}$, n-bromosuccinimide and p-chloromercuribenzoate. Xylobiose (10 mM) was completely decomposed to xylose after 8 hrs enzyme reaction with 2 units of the $\beta$-xylosidase.

• BMB Reports
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• 제35권2호
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• pp.165-171
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• 2002

A serine collagenolytic protease was purified from the internal organs of filefish Novoden modestrus, by ammonium sulfate, ion-exchange chromatography on a DEAE-Sephadex A-50, ion-exchange rechromatography on a DEAE-Sephadex A-50, and gel filtration on a Sephadex G-150 column. The molecular mass of the filefish serine collagenase was estimated to be 27.0 kDa by gel filtration and SDS-PAGE. The purified collagenase was optimally active at pH 7.0-8.0 and $55^{\circ}C$. The purified enzyme was rich in Ala, Ser, Leu, and Ile, but poor in Trp, Pro, Tyr, and Met. In addition, the purified collagenolytic enzyme was strongly inhibited by N-P-toluenesulfonyl-L-lysine chloromethyl ketone (TLCK), diisopropylfluorophosphate (DFP), and soybean trypsin inhibitor.

• Journal of Microbiology and Biotechnology
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• 제6권6호
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• pp.402-406
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• 1996

Inulin fructotransferase (depolymerizing) (EC 2.4.1.93) was purified 34-fold from the culture broth of Arthrobacter sp. A-6 by using a combination of ammonium sulfate fractionation, DEAE-Sepharose CL-6B chromatography and Sephacryl S-200 gel filtration. The purified enzyme converts inulin into di-D-fructofuranose dianhydride III(DFA III) and small quantities of fructo-oligosaccharides. The temperature and pH optima of the enzyme were $70^{\circ}C$ and 6.0, respectively. Molecular weight of the enzyme was determined to be 49 kDa by 12$%$ SDS-polyacrylamide gel electrophoresis, and 145 kDa by Sephacryl S-200gel filtration. This indicates that the functional inulin fructotransferase of Arthrobacter sp. A-6 has a homomeric trimer structure. The enzyme had an isoelectric point of pH 4.6. The N-terminal amino acid sequence of the purified enzyme subunit was Ala-Asp-Asn-Pro-Asp-Gly(\ulcorner)-Ser-Asn-Met(or Glu)-Tyr-Asp-Val.