• 한국미생물·생명공학회지
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• 제23권6호
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• pp.695-701
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• 1995

An alkaline lipase producing bacteria was isolated from soil and identified as Serratia liquefaciens AL-11. from the results of analysis of its morphological, biochemical and physiological properties. This strain showed the highest productivity of alkaline lipase when grown at pH 9.0 and 30$\circ$C for 42 hours in the medium of 1% peptone, 0.5% tryptone, 0.9% yeast extract, 1% starch, 1% tween 80, 0.05% CaCl$_{2}$ and 0.05% NaCl. The enzyme was purified by ammonium sulfate treatment, Sephadex G-100 gel filtration and DEAE-Sephadex A-50 column chromatography. The specific activity of the purified enzyme was 27 unit/mg protein and the yield of enzyme activity was 61.3%. The homogeneity of the purified enzyme was verified by polyacrylamide gel disc electrophoresis. Molecular weight of the purified enzyme was estimated about 53,000 by sodium dodecyl sulfate- polyacrylamide gel electrophoresis. This enzyme is composed of 17 amino acids of which glycine, proline and glutamic acid were three miajor acids.

• Journal of Microbiology
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• 제44권3호
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• pp.276-283
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• 2006

The thermophilic fungus Thermoascus aurantiacus 179-5 produced large quantities of a glucosidase which preferentially hydrolyzed maltose over starch. Enzyme production was high in submerged fermentation, with a maximal activity of 30 U/ml after 336 h of fermentation. In solid-state fermentation, the activity of the enzyme was 22 U/ml at 144 h in medium containing wheat bran and 5.8 U/ml at 48 h when cassava pulp was used as the culture medium. The enzyme was specific for maltose, very slowly hydrolyzed starch, dextrins (2-7G) and the synthetic substrate (${\alpha}$-PNPG), and did not hydrolyze sucrose. These properties suggest that the enzyme is a type II ${\alpha}$-glucosidase. The optimum temperature of the enzyme was $70^{\circ}C$. In addition, the enzyme was highly thermostable (100% stability for 10 h at $60^{\circ}C$ and a half-life of 15 min at $80^{\circ}C$), and stable within a wide pH range.

• BMB Reports
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• 제28권5호
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• pp.408-413
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• 1995

Decrease in the amount of cotyledonary spermidine in Glycine max under anaerobic conditions related to an increase in spermidine dehydrogenase. Under the same conditions, no enzymatic activity of diamine oxidase was observed. Exposure of Glycine max both to spermidine and 1,3-diaminopropane under anaerobic conditions resulted in a decrease in spermidine contents. Correlated with the decrease in spermidine contents, there was a drastic increase in spermidine dehydrogenase. The molecular weight of the purified enzyme estimated by Sephacryl S-300 gel column and SDS gel electrophoresis were 130,000 dalton and 65,000 dalton, respectively, indicating that the enzyme is a dimer. The optimal pH for activity was 9.3. The $K_m$ value for spermidine was 0.61 mM. Neither metal ions nor polyamine and derivatives affected enzymatic activity, but the enzyme was inhibited by DTNB, NEM and PCMB, suggesting that a cysteine residue of the enzyme is associated with or involved in enzyme activity. To our knowledge, this is the first report describing properties of the enzyme from plants. Considered together, the data in this paper indicate that both spermidine and 1,3-diaminopropane, novel activators, enhance the spermidine dehydrogenase activity and control the intracellular spermidine contents.

• Applied Biological Chemistry
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• 제12권
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• pp.7-11
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• 1969

Mucor-rennin, the crystalline milk-clotting enzyme, isolated from Mucor pusillus var. Lindt, has an acid protease activity. The optimum pH for the digestion of k-casein is 4.5, while that for hemoglobin digestion is 4.0. The skim milk solution was easily clotted acidic solution than alkalin solution, and the milk clotting activated by Ca ion. The enzyme was heat stable against heat from pH 4.0 to 6.0 but was more stable at pH 5.0. The activity of the enzyme at pH 5.0 did not decrease at 30 C for 15 days and the activity was not effected by sodium propionate and salicilic acid. Therefore, the enzyme of liguid type could store for a long time and could be transported from Erzyme production Co. to Manufacture of cheese Co. by adding the antiesptic and by adjusting pH to 5.0.

• Journal of Microbiology and Biotechnology
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• 제8권3호
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• pp.270-276
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• 1998

Extracellular ${\alpha}$-glucosidase was purified to homogeneity from moderately thermophilic Bacillus sp. DG0303. The thermostable ${\alpha}$-glucosidase was purified by ammonium sulfate fractionation, ion-exchange chromatography, preparative polyacrylamide gel electrophoresis (PAGE), and electroelution. The molecular weight of the enzyme was estimated to be 60 kDa by SDS-PAGE. The optimum temperature for the action of the enzyme was at $60^{\circ}C$. It had a half-life of 35 min at $60^{\circ}C$. The enzyme was stable at the pH range of 4.5~7.0 and had an optimum pH at 5.0. The enzyme preparation did not require any metal ion for activity. The thermostable ${\alpha}$-glucosidase hydrolyzed the ${\alpha}$-1,6-linkages in isomaltose, isomaltotriose, and panose, and had little or no activity with maltooligosaccharides and other polysaccharides. The $K_m$ (mM) for p-nitrophenyl-${\alpha}$-D-glucopyranoside (pNPG), panose, isomaltose, and isomaltotriose were 4.6, 4.7, 40.8, and 3.7 and the $V_{max}$(${\mu}mol{\cdot}min^-1$$mg^-1$) for those substrates were 5629, 1669, 3410, and 1827, respectively. The N-terminal amino acid sequence of the enzyme was MERVWWKKAV. Based on its substrate specificity and catalytic properties, the enzyme has been assigned to be an oligo-1,6-glucosidase.

• 한국미생물·생명공학회지
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• 제17권1호
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• pp.69-73
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• 1989

자가분해 효소는 Cl. butyricum의 포자형성 배지에서 배양할 때 체외로 배출되어 배양액에도 존재하였다. 배양액으로부터 약 50배로 부분정제된 자가분해 효소를 사용하여 효소의 성질을 조사하였다. 자가분해 효소의 최적 pH와 온도는 각각 5.0과 37$^{\circ}C$였으며 중성 pH에서는 안정하나, 열에는 비교적 불안정하여 5$0^{\circ}C$에서 5분간 열처리한 후 효소활성의 70%가 소실되었다. 또한 Cu ion$^{++}$에 의해서 효소활성이 저하되었으나 그 밖의 금속이온에 의하여서는 큰 영향을 받지 않았다. 또한 자가분해 효소는 기질로서 영양세포에는 직접 활성을 나타내지 못하나, 세포벽 fraction에는 활성을 가지고 있었다.

• 한국식품영양학회지
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• 제16권2호
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• pp.152-157
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• 2003

Serratia marcescen ATCC 25419 protease를 ammonium sulfate treatment, DEAE-cellulose anion exchange chromatography등의 방법으로 정제하였는데 최종 단계에서 667.5 unit/mg 이었으며 회수율은 43%이었고 448배 정제되었다. 정제한 S. marcescens protease로부터 아포효소를 만든 후 금속 재활성화에 대해 조사하였다. S. marcescens protease는 EDTA에 의해 완전히 활성을 잃는 metalloenzyme이며 Hg, Fe, Cu 등에 의해서 효소 활성을 70% 이상 잃은 반면, Co는 효소 활성을 약 20% 정도 증가시켰다. 아포효소의 재활성화는 pH 6~8에서 Mn, Co, Zn 등이 효과적이었다. Mn, Co, Zn등을 아포효소에 가하여 만든 효소들 중에서 Zn-효소는 효소 활성도, 알칼리-불활성화, 열-안정성 면에서 원래 protease와 유사하였다.

• Journal of Dairy Science and Biotechnology
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• 제26권2호
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• pp.39-43
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• 2008

EMC는 치즈와 유사한 효소작용은 거치게 되지만 치즈에 비하여 더 많은 효소작용에 의하여 더욱 강력한 풍미를 생산하게 된다. 이러한 우수한 EMC의 개발을 위하여 적합한 효소를 찾아내는 것은 중요한 과제이다. 심지어 동일한 PGE도 송아지의 것이 어린 염소나 어린 양의 것에 비하여 더 부드러운 맛을 생산할 수 있다. 특히 체다치즈의 풍미를 위하여는 미생물효소보다 동물성 esterase나 peptides가 유리한 것으로 알려지고 있으며, 체다치즈나 스위스치즈의 경우 비단 백태 질소화합물의 양도 EMC가 치즈에 비하여 3배 가까이 많았다. 체다 EMC의 성분 비율에서도 유리지방산에는 butyric acid, myristic acid, palmitic acid와 oleic acid가 많았고, 유리 아미노산에는 glutamic acid, valine, leucine과 lysine이 많은 것으로 나타났다.

• 미생물학회지
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• 제26권4호
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• pp.368-374
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• 1988

Bacillus polymyxa YL38-3이 생성하는 세포의 cytosine deaminase의 효소학적 ,성질을 검토하였다. 본 세포외 효소는 열 안정성이 높으며, 인산완충액(pH6.0)과 $30^{\circ}C$에서 효소활성이 최대를 나타냈다. 본 효소는 cytosine 뿐 아니라 5- fluorocytosine-을 기질로 하나, 5-methylcytosine은 촉매하지 않았다. 더우기 본 효소는 $Cd^{2-}$, $Hg^{2+}$의 중금속이온과 ImM p-chloromercuribenzoate에 의하여 완전히 실활되며, o-phenanthroline과 monoiodoacetate에 의하여 75% 저해되었다. 그러나 1mM 2-mercaptoethanol에 의하여 본 효소의 활성을 약 200% 이상 활성화시켰다.

• 한국미생물·생명공학회지
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• 제11권1호
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• pp.15-21
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• 1983

전보에서 분리 동정한 Y-33 균주의 $\beta$-galactosise 효소생산조건을 검토하고 효소를 정제하여 정제효소의 성질을 조사한 결과는 다음과 같다. 1. 효소생산을 위한 최적초발 pH는 7.0이었고 최적온도는 $65^{\circ}C$이었다. 2. 효소는 lactose와 galactose에 의하여 유도되어졌으며 세포내효소이었다. 3. 조효소액을 1차 DEAE-cellulose, 2차 DEAE-cellulose column chromatography 및 Sephadex G-150로 gel filtration하여 정제도가 28.3배, 수율이 15.2%의 정제효소를 얻었다. 4. 정제효소는 polyacrylamide gel 전기 영동에 의하여 순도가 검정되었다. 5. 정제효소의 유당가수분해를 위한 최적작용 온도는 $65^{\circ}C$. pH는 6.5이었다.