• Biomedical Science Letters
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• v.6 no.3
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• pp.209-221
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• 2000

The purpose of this study was to investigate the practical availability of pretense production that can be used at home after isolating Serratia sp.2000-1 which produced extracellular pretense from clinical specimen. Basic production conditions and partial enzymatic characteristics of pretense produced by Serratia sp. 2000-1 was as follows: The kind and concentration of carbohydrate, nitrogen and metal salts for optimal enzyme production condition were each identified as the concentration of 1.5% glucose, 2.0% CSP, and 0.1% CaCl$_2$, and the optimal temperature, time and initial pH for culture were each 3$0^{\circ}C$, 72 hours, and pH 8.0. The final enzymatic yeild that was purified by 3 steps with ammonium sulfate precipitation (45~80%), DEAE-cellulose column chromatography, and Sephadex G-200 gel chromatography was 14.4%, and enzyme inactivity rate increased approximately 291314s. The optimal temperature and pH for purified pretense activity were 35$^{\circ}C$ and pH 7.0~8.0, and purified pretense activity was relatively stable by 4$0^{\circ}C$ at pH 6~10 for 30 min, however heating at 6$0^{\circ}C$ for 30 min, it liminated detectable pretense activity. The pretense activity was activated by $Mg^{2+}$, $Ba^{2+}$, $Ca^{2+}$, Mn$^{2+}$, but inactiviaed by Hg$^{2+}$, Ag$^{2+}$, Cu$^{2+}$, and the pretense activity was inhibited strongly by SDS among enzyme activity inhibitors. Further study is required to evaluate the practical availability of pretense production that can be used at home by isolating Serratia sp. from more clinical specimen and examining pretense more in details.

• The Korean Journal of Mycology
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• v.15 no.3
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• pp.175-182
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• 1987

A raw starch saccharifying enzyme from Aspergillus sp. SN-871 was purified by ammonium sulfate precipitation, DEAE-cellulose column chromatography, CM-Sephadex C-50 column chromatography and Sephadex G-75 gel filtration. The specific activity of purified enzyme was 18 fold and the yeild was 13.40%. The molecular weight of the purified enzyme was estimated as approximately 40,000 dalton by the method of Andrews gel filtration. The optimum pH and temperature for this enzyme were found to be 4 and $40^{\circ}C$, respectively and the stable range of pH was 2 to 5. The enzyme was themostable at below $60^{\circ}C$ and inactivated at $70^{\circ}C$. It showed a tendency to increase the enzyme activity under the presence of 0.01 M $BaCl_2$, but under 0.01 M$Pb(NO_3)_2$, $AgSO_4$, and $K_3Fe(CN)_6$ and citric acid etc. inhibited it completely. The substrate specifity of enzyme showed a tendency to increase the enzyme activity under addition of dextrin and glycogen, but under saccharose inhibited it. COD removal rate of Aspergillus sp. SN-871 was approximately 67 to 68%.

• Korean Journal of Food Science and Technology
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• v.30 no.1
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• pp.82-89
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• 1998

This study was performed to elucidate the purification and characterization of pretease from saewoo-jeot, a Korean traditional salt-fermented shrimp product. The protease in saewoo-jeot (Acetes japonicus) were extracted, desalted through electrodialysis and purified by ammonium sulfate fractionation, Sephadex G-100 gel filtration and DEAE-cellulose column chromatography. Purified enzyme had specific activity of 8.4 unit/mg, yield of 14% and purification fold of 9.8. Purified enzyme was confirmed as single band protein by polyacrylamide gel electrophresis and the molecular weight was estimated to be about 24 kDa. The optimal pH and temperature for the enzyme activity were 8.0 and $40^{\circ}C$, respectively. The range of its stability to the pH and temperature were 7.0 to 10.0 and $30^{\circ}C\;to\;60^{\circ}C$, respectively. The activity of enzyme to synthetic substrate showed BAPNA and TAME. The enzyme was activated significantly by manganese ions, while inhibited by STI, TLCK. metals $(K^+,\;Li^+,\;Na^+,\;Ca^{++},\;Co^{++},\;Cu^{++},\;Mg^{++},\;Ba^{++},\;Hg^{++},\;Zn^{++},\;Fe^{+++})$. The Km value of the enzyme was $5.1{\times}10^{-7}\;M$ to hammersten casein. It's suggested that purified protease from saewoo-jeot seemed to be trypsin-like enzyme.

• Journal of the Korean Society of Food Science and Nutrition
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• v.23 no.4
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• pp.604-613
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• 1994

Hydrophilic brown pigments(HBPs) produced during fermentation and aging the domestic Meju and Doenjang were separated by dialysis and chromatography , and their antioxidant activity were measured . The chemical properties of HBP were determined by UV and IR spectrophotometry. HBPs contents were found to be 93.1 mg/g and 183.2mg/g in Meju fermentated for 30 days and 80 days , respectively. The ratio of dialysate to diffusate of the HBPs were appeared to be 70 : 30 and 87 :13 in the Meju fermented for 30 days and 80 days, respectively. and the rtio in the Doenjang aged for 60 days was 91 :9 , indicating that dialysate slowly days, respectively, and the ratio in the Doenjang aged for 60 days was 91 : 9, incidatin that dialysate slowly increased by the fermentation . Both portion exhibited strong antioxidant activity, but more stronger antioxidant activity was found in the dialysate. DEAE-celluose column chromatography showed that dialysate contained more materials eluted by 0.01-0.03M HCI solution than 0.01M acetate buffer, but diffusate showed opposite results. The degree of browning reaction and antioxdiant activity found in the fraction eluted by HCI solution was stronger than that of the fraction eluted by acetate buffer. The fraction eluted from DEAE-cellulose column chormatogrphy was further fractionaged by TLC and found that strong antioxidant activity was present in the fractions which did not possess fluorescenece and showed a negative ninhydrin reaction. TLC fractions of HCI eluant in Meju exhibited a strong absorbance at 260-280nm, but most of other fractions did not show any absorbance at UV region. TLC fractions from dialysate and diffusate showed fairly identical IR spectrum with absorbance at 3400cm-1, 2800cm-3000cm -1, 1600cm, -1 1400cm-1 and 1100 cm-1 , however, in addition to these absorbances, the spectrum from HCI eluant of Meju exhibited a strong absorbance at 1750cm , indicating the carbonic acid or carbonate ester.

• Fisheries and Aquatic Sciences
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• v.9 no.3
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• pp.115-117
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• 2006

A micro algal growth-enhancing polysaccharide compound was isolated from the green alga Monostroma nitidum using water extraction, molecular fractionation, a DEAE-cellulose column, and fast protein liquid chromatography using a Superose-12 column. The yield of the compound from the seaweed powder was 8.3$\times$l0$^{-3}$%. At 2 mg/mL concentration, the polysaccharide enhanced Tetraselmis suecica cell growth in f/2 medium by approximately 160%.

• Microbiology and Biotechnology Letters
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• v.24 no.3
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• pp.316-320
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• 1996

An alkaline proteinase produced by Yarrowia lipolytica TH65 was purified by 40-65% ammonium sulfate fractionation, DEAE-cellulose chromatography, and gel filtration with Sephadex G-100 and Sephadex G-75. The purified enzyme was shown as a single band on SDS-PAGE, and its molecular weight 31,500. Optimum temperature and pH were 40$\circ$C and 8.5-9.0, respectively, and the enzyme was stable below 40$\circ$C and in the pH range of 6-8. The enzyme was strongly inhibited by divalent ions, completely by PMSF, and partially by EDTA, EGTA, and phenanthroline. But the inhibitory effect in the presence of EDTA, EGTA and phenanthroline could be reversed by addition of Ca$^{2+}$. Thus, these results indicated that the purified enzyme was an alkaline serine proteinase (E.C. 3.4.21.14).

• Korean Journal of Food Science and Technology
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• v.23 no.4
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• pp.394-399
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• 1991

The strain OK-63 isolated from katsuobushi was cultured on wheat bran medium and the isolate was morphologically identified as an Aspergillus niger group and showed maximum pretense activity and multiplication after 6 days of cultivation. Protease was purified by ammonium sulfate fractionation. Sephadex G-100 gel filtration and DEAE-cellulose column chromatography. The purified enzyme showed single band by polyacrylamide gel electrophoresis and the purity was 150 times higer than crude enzyme. The recovery of enzyme activity was found to be 45%.

• Journal of environmental and Sanitary engineering
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• v.5 no.1
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• pp.49-64
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• 1990

Glucoamylase from the culture filtrate of Aspergillus nicer was purified by ammonium sulfate precipitation, aceton precipitation, DEAE-cellulose ion exchange chromatography and Sephadex G-50 gel fillration. Glucoamylase was secreted into the medium upon growth on glucose, sucrose or a variety of other hexose sugars or hexose sugar polymers and little or no glucoamylase activity was found when glycerol or xylose was used as the carbon source. The optimum pH and temperature (or the maximum enzyme activity were found to be 5.0 and $50^{\circ}C$, respectively. The enzyme was considerably thermostable, for no loss of activity was observed when the enzyme was preincubated at $60^{\circ}C$ for 30 min. The enzyme activity was inhibited by 20 mM of $Hg^{2+}$, $Fe^{2+}$. The km value for starch was 0.045%.

• Journal of Plant Biology
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• v.37 no.2
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• pp.151-158
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• 1994

Aleurone layers of normal and vp1 mutant maize kernels were extracted and centrifuged at 100,000g to yield a cytosol fraction. Binding of [3H]ABA cis, trans (＋)ABA to a soluble macromolecular components present in the cytosol was demonstrated by Sephadex chromatography and non-denaturing PAGE. The binding component was of high molecular weight and seems to be an aggregate of proteins. A rapid DEAE-cellulose filter method for assaying bound [3H]ABA to a soluble protein was adapted. Binding assays were performed with cytosol that had been preheated or incubated with several enzymes, indicating that heat and protease treatments disrupted the binding. This suggested that binding occurred to proteins. Some properties of the ABA binding proteins were described. The [3H]ABA binding were reduced dramatically when unlabeled ABA was added as a competitor, suggesting a specific binding of [3H]ABA. Gel filtration profiles and autoradiogram of [3H]ABA binding showed no difference in the binding components of Vp1 and vp1/vp1 mutant cytosol, indicating that Vp1 protein is not a sole ABA binding protein.

• Korean journal of food and cookery science
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• v.10 no.4
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• pp.339-345
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• 1994

Polyphenol oxidase in Yam was partially purified through ammonium sulfate fractionation, DEAE-cellulose column chromatography. The specific activity of purified PPO was 138.22 unit/mg protein. The optimum pH and temperature of purified PPO were respectively 7.0 and 30$^{\circ}C$. The heat treatment at 80$^{\circ}C$ for 6 min, decreased PPO activity to 50%. The enzyme showed high substrate specificity toward catechol. The Km value for catechol was 5 mM. In the Ames test using S. typhimurium TA98 and TA100 catechol-YEBRP, pyrogallol-YEBRP, chlorogenic-YEBRP showed strong antimutagenicity on sodium azide and MECF excepting hydroquinone-YEBRP showed killing effect on both strains.