• Food Science and Biotechnology
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• v.15 no.2
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• pp.177-182
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• 2006

Polyphenol oxidase (PPO) was isolated from the flesh of Fuji apples by DEAE-Cellulose, ammonium sulfate precipitation, phenyl-Sepharose CL-4B, and Sephdex G-100 chromatography. The molecular mass of the purified PPO was estimated to be 40 kDa by SDS polyacrylamide gel electrophoresis. With regard to substrate specificity, maximum activity was achieved with chlorogenic acid as substrate, followed by catechin and catechol whereas, there was no detectable activity with hydroquinic acid, resorcinol, or tyrosine as substrate. The optimum pH and temperature with catechol as substrate were 6.5 and $35^{\circ}C$, respectively. The enzyme was most stable at pH 6.0 and unstable at acidic pH. The enzyme was stable when it was heated to $45^{\circ}C$ but heating at $50^{\circ}C$ for more than 30 min caused 50% loss of activity. Reduced $ZnSO_4$, L-cystein, epigallocatechin-3-o-gallate (EGCG), and gallocatechin gallate (GCG) also inhibited activity.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2002.04b
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• pp.27-31
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• 2002

Samyang Genex succeeded in commercialization of anticancer agent-paclitaxel by plant cell culture technology. The core technology of Samyang Genex relating paclitaxel production includes cell line development, cell line preservation, cell culture, scale-up technology, and purification technology. On the basis of the research, Samyang Genex built the factory operated by CGMP (current good manufacturing practice). The $paclitaxel-Cenexol^{TH}-is$ commercially available in Korea, and it will be launched to world market including USA after approval of US FDA.

• Journal of Plant Biotechnology
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• v.4 no.1
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• pp.39-44
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• 2002

The fast-migrating cationic peroxidase isozyme, named RC3, was purified from rice (Oryza sativa cv. Nak-Dong) callus. Purification of the enzyme was accomplished by ammonium sulfate fractionation, CM-cellulose ionexchange chromatography, and Sephacryl S-100 gel filtration. The molecular mass of the enzyme was about 34 KDa as determined by SDS-PACE and 38 KDa by Sephacryl-100 gel filtration. The pI value of the enzyme was 8.9. Antiserum against RC3 was raised in rabbits, and anti RC3 antiserum reacted with RC3 isozyme by Ouchterlony double immunodiffusion. The optimum pHs and Km values of the enzyme for various substrates were determined. Kinetic studies with various substrates showed that RC3 had very low Km value of 0.01 mM for ferulic acid and ascorbic acid. However, the enzyme did not use esculetin as a substrate.

• Food Science and Biotechnology
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• v.14 no.5
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• pp.581-585
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• 2005

The antimicrobial activity of partially purified enterocin K25, produced by Enterococcus sp. K25, was abolished by proteases such as pepsin and proteinase K. The bacteriocin was resistant to heat treatment at $75^{\circ}C$ for 15 min and lost 75% of its activity at $100^{\circ}C$ for 30 min. Enterocin K25 showed bactericidal mode of action against an indicator strain, Lactobacillus plantarum NCDO 955. Enterocin K25 was purified to 112.6-fold purity via conventional steps of ammonium sulfate precipitation, ion exchange chromatography, and reversed phase high performance liquid chromatography (RP-HPLC). The molecular mass of the purified enterocin K25 was estimated as 4.3 kDa on an electrophoresis gel. Plasmid (${\sim}6.5\;kb$) linkage of production of enterocin K25 was confirmed by plasmid curing.

• Journal of Marine Bioscience and Biotechnology
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• v.2 no.4
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• pp.245-249
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• 2007

High quality of purified alginate and fucoidan is required for the medical uses to prevent the unexpected side effects from the contaminated endotoxin present in the materials. We attempted to establish an efficient and fast mass scale production method for the highly purified poly-G alginate from the sea weeds. In addition, lab scale method was established to obtain the fraction of higher purity from the commercially available fucoidan source (about 85% purity).

• Journal of Microbiology and Biotechnology
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• v.26 no.2
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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.

• Journal of the Korean Society of Food Science and Nutrition
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• v.28 no.4
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• pp.816-821
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• 1999

Cholesterol esterase(pCEH, pancreas cholesterol ester hydrolase, E.C.3.1.1.13) which is secreted from pancreas has been known as an important lipase for cholesterol uptake. cholesteryl acyl esters from a diet must be hydrolyzed to free cholesterol and fatty acid by cholesterol esterase before the absorption in small intestine. For the development of inhibitory substances from natural source, we screened many extracts of oriental herbs for the inhibition of cholesterol esterase in vitro. The ethanol extract of Ephedra herba showed strong inhibitory activity. Solvent fractionation and silica gel column chromatography with the extract lead to the purification of the inhibitory principle in Ephedra herba. Crystallized inhibitor was identified as ( ) ephedrine by using UV, FT IR, 1H NMR, 13C NMR and GC/Mass. These results suggest that ( ) ephedrine can be used as a potential lead compound for the development of inhibitor for cholesterol uptake by cholesterol esterase inhibition.

• BMB Reports
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• v.35 no.2
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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.

• Microbiology and Biotechnology Letters
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• v.28 no.5
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• pp.258-263
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• 2000

A fibrinolytic enzyme was purified to homogeneity from Bacillus sp. S19 using DEAE and CM column chromatograhies, and gel filtration with a recovery yield of 13%. Its molecular mass was estimated to be 42 kDa by SDS-PAGE. The pH and temperature optima were 8.0 and $40^{\circ}C$, respectively. The enzyme was stable up to $45^{\circ}C$ and over a pH range of 6-9. The N-terminal amino acid sequence of the enzyme was determined as Alsa-Gln-Asp-Ala-Thr-Val-Asn-Ile-Ser-Ala-Glu-Arg-Gln-Val-Ile. The fibrinolytic activity was increased by $Cu^{2+}$ while it was strongly inhibited by metal ions such as $Cd^{2+}$ and $Ba^{2+}$ . In addition, the enzyme was inhibited by EDTA, but not by PMSF, suggesting that it is a metallorprotease.

• Journal of Microbiology and Biotechnology
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• v.8 no.3
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• pp.251-257
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• 1998

Cycloinulooligosaccharide fructanotransferase (CFTase) which produces cyclofructan from inulin was purified 332-fold from a culture broth of Bacillus macerans CFCl. The molecular mass of the CFTase was estimated to be 110 kDa by SDS-polyacrylamide gel electrophoresis and gel filtration, indicating that the enzyme has a monomer structure. The maximal level of enzyme activity was observed at pH 7.5 and $45^{\circ}C$. The enzyme was stable in the pH range 6.0 to 9.5, and at temperatures up to $45^{\circ}C$ for 1 h. The enzyme activity was completely inhibited in the presence of 0.5 mM $Ag^+\;or\;Cu^2+$ ion. None of sucrose (GF), l-kestose (GF2), or nystose (GF3) were found to be substrates for the CFTase, but inulooligosaccharides larger than nystose were attacked by the enzyme. The CFTase catalyzes not only the cyclization as the major reaction, but also disproportionation and coupling reactions involving intermolecular transfructosylation in the same manner as cyclodextrin glucanotransferase (CGTase) (EC 2.4.1.19).