• Korean Journal of Microbiology
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• v.30 no.4
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• pp.291-298
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• 1992

Srrepromycec. corlirolar produces at least 4 catalase activity bands with different electrophoretic mobilities on polyacrylamide gel which vary during development. Spores and mycelia at stationary phase produced all the activity bands(Cat1. 760 kr); Cat3-I, 170 kD: Cat3-2, 140 kD: Cat3-3. 130 kD; Cat4, 70 kD) except for Cat2 (300 kD). Mycelia at mid-logarithmic phase produced only Cat2 and Cat3-2 bands, and mycelia at late-logarithmic phase produced bands except Catl and Cat\ulcorner. Catalase-deficient mutants were screened in S. coelicalur by H201 bubbling test following NTG mutagenesis. Wc tested sevcral non-bubbling or slow-bubbling mutants for their catalase activities. The overall activities in cell extracts decreased more than 5 fold. Activity bands in native gel selectively decreased in intensity or disappeared. In all the non-bubbling mutants testcd, Cat3-2 band decreased significantly or disappeared. suggesting that Cat3-2 is the major catalase. The selective disappearance of bands in mutants suggest that each band is governed by different genes. We purified catalase activity from -:ell extracts obtained at late-logarithmic phase. Following chromatographies on Sepharose CL-4B. DEAE Sepharose CL-6B. Phcnyl Sepharose CL-4B. and hydroxylapatite columns. only the Cat3-2 activity was obtained. The native form of Cat3-2 has molecular weight of approximately 140 kD, judged by gel electrophoresis. Thc electrophoretic mobility on SDS-polyactylamide gel suggests that this enzyme contains 2 identical subunits of 67 kD.

• The Korean Journal of Mycology
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• v.40 no.2
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• pp.109-113
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• 2012

We have isolated an anti-complementary polysaccharide from the hot water extracts of Phellinus linteus mycelia. Anti-complementary polysaccharide, PL-5-IIIa, was purified by ultrafiltration, gel permeation chromatography using Sepharose CL-4B. GPC (Sepharose CL-4B) and its homogenicity was demonstrated by HPLC. Using gel permeation chromatography with standard dextrans, its molecular weight was determined as about 800,000 dalton. The purified PL-5-IIIa was identified as a protein bound polysaccharide comprising of 29.6% protein and 64.2% carbohydrate which was composed of fucose(15.8%), galactose(43.1%) and mannose(40.6%).

• Food Science and Preservation
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• v.10 no.2
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• pp.246-251
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• 2003

Pretense produced by Bacillus subtilis PANH765 was purified from culture supernatant by using ammonium sulfate fractionation DEAE-cellulose ion exchange chromatography, and gel filtration with Sephacryl S 200 HR and Sepharose CL-6B. DEAE-cellulose ion exchange column chromatography, separated the pretense into one fraction. This fraction was further purified using Sephacryl S 200 HR and Sepharose CL-6B gel titration. The molecular mass of pretense was estimated to be 35.0 kDa by the SDS-PAGE and gel filtration using Sepharose CL-6B. The results indicated that the purified pretense are monomeric proteins. Specific activity and purification folds of pretense were 657 U/mg and 4.35, respectively. The optimum temperature, optimum pit stable at a temperature range and pH ranges for the purified protease were 65$^{\circ}C$, 7.05, 50 ∼ 75$^{\circ}C$ and 6.0 ∼ 7.5, respectively. The pretense activity was decreased by the presence of PMSF and DFP, which the protease activity was increased by the presence of Na$\^$+/, K$\^$+/, Mg$\^$2＋/ and NH$_4$$\^$+/ ions.

• KSBB Journal
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• v.14 no.2
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• pp.192-197
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• 1999

marine bacterium Pseudomonas sp. CHCS-2 produced the biosurfactant in the culture broth which contained 2%(w/v) arabian light crude oil and the productivity of biosurfactant was increased with the addition of glucose. The crude oil in the culture broth was degraded by this strain and carbon chain of $_nC_{12}~_nC_{22}$ was completely degradaded during the incubation for 196 h. The crude biosurfactant was purified by Amberlite XAD-7, Sepharose CL-4B and DEAE-Sepharose CL-6B column chromatography. Therefore, 0.21g/L of the purified biosurfactnat was obtained. The purified biosurfactant was a type of lipoprotein and the molecular weight was estimated as 67kDa by SDS-PAGE. The lipid composition was identified as octadecanoic acid by gas chromatography/mass spectrometry. And then, the N-terminal amino acid sequence of the protein was determined as Ser-Val-lle-Asn-Thr-lle-X-Met-lle-Gly-Gln-Gln- and the sequence did not show homology to any other known lipoprotein. Therefore, the purified lopoprotein was predicted novel biosurfactant.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.2
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• pp.219-222
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• 2005

Some structural characteristics of hot-water soluble and insoluble starches (waxy black rice starch) were investigated. The hot-water soluble material content of waxy black rice starch was higher (16.6%) than that (13.4%) of Shinsunchalbyeo starch heated at 98$^{\circ}C$ for 8 min. The Amax and absorbance at 625 nm for hot-water soluble and insoluble material of waxy black rice starch were lower than those of Shinsunchalbyeo. Elution patterns of hot water soluble and insoluble materials by gel permeation chromatography (Sepharose CL-2B) were similar in both samples.

• KSBB Journal
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• v.15 no.3
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• pp.318-322
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• 2000

An oxygen-sensitive fumarate reductase has been purified from the cytosol fraction of the Enterococcus faecalis RKY1 grown anaerobically on a defined medium containing glycerol and fumarate. A major portion of the purification was performed with employing Triton X-100 and reducing agents by Phenyl-sepharose CL-4B DEAE-sepharose and Dephadex G-150 The final activity was 0.42 unit/mg. The deduced molecular mass of active band was 66 kDa. The optimal pH and temperature for the activity were 7.0 and 38$^{\circ}C$ respectively. The enzyme activity was not affected by 1mM metal ions such as bacl2 $.$2H2O HgCl2 MnCl2$.$4H2O ZnCl2 CuCl2$.$2H2O Mgcl2$.$6H2O FeSo4$.$7H2O and by EDTA. Partially purified enzyme ws yellow in color ; spectroscopic study indicated the presence of flavins as a cofactor.

• Journal of the Korean Wood Science and Technology
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• v.19 no.2
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• pp.22-29
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• 1991

The endo-1,4-${\beta}$-xylanase was extracted and purified from the extracellular xylanolytic systems of Trichoderma viride. The crude enzyme was chromatographed with ion-exchange reins of DEAE Sepharose CL-6B, Sepharose, S-Sepharose CL-6B and the resulting xylanase was turned out to be a single protein as 20KD hy SDS-polyacrylamide gel electrophoresis. The xylooligomers were obtained from xylan by incubation with the purified xylanase up to 50%. The ${\beta}$-xylosidase lost its activity completely by incubation of crude enzyme for 24hr with buffer solution of pH 2.8 at $27^{\circ}C$. And also, the xylooligomers were obtained from xylan as a main product by incubation with the crude enzyme treated with acidic buffer.

• Korean Journal of Microbiology
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• v.26 no.1
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• pp.52-59
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• 1988

A partially purified preparation of L-galactonolactone oxidase which catalyzes the last step of L-ascorbic acid biosynthesis was obtained from Saccharomyces cerevisiae ATCc 26787. The purification procedures included Triton X-100 treatment, protamine sulfate precipitation, ammonium sulfate precipitation, DEAE-Sepharose CL-6B ion exchange chromatography, Sephadex G-150 gel filtration chromatography, and Phenyl-Sepharose CL-4B hydrophobic interaction chromatography. The optimum temperature for the enzyme activity was about $34^{\circ}C$ and the optimum pH was 6.8-7.0. The substrate specificity was confined to L-aldonolactones, L-galactono-1,4-lactone and L-gulono-1,4-lactone. An apparent Km value of 0.294mM with L-galactono-1,4-lactone as a substrate was found. By comparing the substrate specificities of this enzyme with those of isofunctional enzymes of higher plants and animals, it becomes evident that the enzyme of S. cerevisiae ATCC 26787 is rather similar to the L-gulonolactone oxidase of animals than the galactonolactone dehydrogenase of higher plants.

• Journal of environmental and Sanitary engineering
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• v.12 no.2
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• pp.59-64
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• 1997

For purification of a 70kDa toxic protein of mosquitocidal delta-endotoxin from B. thuringiensis strain H9B, immuno-affinity chromatography was performed. After separation of 70kDa toxic proteins from the delta-endotoxin of the strain H9B on SDS-PAGE, the 70kDa toxic protein was subcutaneously injected into rabbit for making a polyclonal antibody. A anti-70kDa toxic protein was purified by a column chromatography packed with protein A-sepharose 4B gels. The 70kDa toxic protein from delta-endotoxin of the strain H9B was also purified by an immuno-affinity chromatography packed with CNBr-activated sepharose 4B gels conjugated anti-70kDa toxic protein after elution with 1/10M citric acid-1/5M Na$_{2}$HPO$_{4}$ buffer(pH3.2) containing 0.5M NaCl. The 70kDa toxic protein was purified through only one step-separation system, was demonstrated by SDS-PAGE and immunoblot.

• Korean Journal of Food Science and Technology
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• v.27 no.5
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• pp.673-679
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• 1995

Pectinesterase(PE) has been definitively established as the causative agent for clarification of citrus juice and gelation of frozen concentrates. This enzyme is present in multiple forms in citrus fruit. Although representing a minor fraction of the total PE activity, thermostable pectinesterase(TSPE) accounts for the severe time/temperature processing treatment required to inactivate PE. This study was undertaken to separate and purify thermolabile pectinesterase(TLPE) and TSPE from the crude PE extracts of Valencia orange rag-pulp powder. The approach taken was to carry out the three kind of chromatographies of CM-Sephadex C-50, Phenyl Sepharose CL-4B and CM-Biogel A to the unheated crude PE or the heated crude PE. All of them used could increase the purity of PEs and, specially, Phenyl Sepharose CL-4B chromatography could separate crude PE as the mixture of PEs into two forms of TLPE and TSPE. The purified TLPE had specific activity of 1,005 units/mg, yield of 13.6% and purification of 35 fold, while the TSPE separated from the unheated crude PE showed specific activity of 3,115 units/mg, yield of 1.5% and purification of 100.5 fold, and another TSPE from the heated crude PE was found to be specific activity of 1,803 units/mg, yield of 15.4% and purification of 140 fold.