• Korean Journal of Fisheries and Aquatic Sciences
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• 제24권5호
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• pp.273-288
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• 1991

To elucidate the physiological and biochemical differences between chondrichthyes and osteichthyes, the properties of the specific digestive enzymes in cat-shark, Cephaloscyllium umbratile, and mackerel, Scomber japonicus, were studied. Homogenous trypsin proved through the disc-electrophoresis, SDS-PAG electrophoresis and gel filtration was obtained from the pancreas of cat-shark by $50-70\%$ saturated ammonium sulphate fractionation, DEAE-Sephadex A-50 column chromatography, benzamidine-Sepharose 6B affinity chromatography and Sephadex G-75-120 gel filtration. Two types of trypsins were also obtained from the pyloric caeca of mackerel by $30-70\%$ saturated ammonium sulphate fractionation and the slightly modified procedure from the method adopted in the purification of cat-shark trypsin. The two trypsins, designated trypsin A and B, were proved their homogeneity by disc- and SDS-PAG electrophoresis and gel filtration. The molecular weights of the trypsins were estimated to be 31,700 for cat-shark trypsin, 30,000 for mackerel trypsin A and 29,000 for mackerel trypsin B by SDS-PAG electrophoresis, but those were estimated to be 21,500 for cat-shark trypsin, 23,700 for mackerel trypsin A and 21,500 for mackerel trypsin B by gel filtration. The trypsins exhibited their optimum conditions at pH 9.0 and on temperature ranged from $45^{\circ}C\;to\;50^{\circ}C$ for cat-shark, and at pH 8.0 and a temperature of $50^{\circ}C$ for mackerel trypsin A and B, respectively. The cat-shark trypsin was stable at pH 10.0 and the temperature below $10^{\circ}C$, whereas the mackerel trypsin A and B, were stable in the range over pH 7.0 to pH 9.0 below $10^{\circ}C$ and at pH 8.0 below $35^{\circ}C$, respectively. The mackerel trypsins were severely inhibited by some heavy metal ions such as $Ag^{2+},\;Cu^{2+}\;and\;Hg^{2+}$ compared to cat-shark trypsin. All of the enzymes were also inhibited by antipain, leupeptin, TLCK(tosyllysine chloromethyl ketone) and SBTI(soybean trypsin inhibitor) remarkably. The inhibitory effects of PMSF(phenylmethane sulphonylfluoride), DFP(diisopropyl fluorophosphate) and benzamidine were indicated that these enzymes belong to serine-proteases.

• Korean Journal of Food Science and Technology
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• 제30권3호
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• pp.630-637
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• 1998

Enzymatic properties of immobilized transglucosidase (TG) from Aspergillus niger was characterized and compared with soluble TG. Michaelis-Menten constant $(K_m)$ and maximum velocity $(V_{max})$ of immobilized TG were $122\;mM,\;3.9{\times}10^{-2}\;mM/min$ and in case of soluble TG, 21 mM, 0.4 mM/min. The optimum pH of immobilized TG was pH 5.0 like soluble TG but immobilized TG showed 16% and 45% higher activity than soluble TG at pH 5.0 and pH 6.0. Both of immobilized TG and soluble TG were stable from pH 2.0 to pH 9.0, and therefore their activities in these pH ranges were remained more than 90%. The temperature was optimal at $60{\sim}70^{\circ}C\;and\;70{\sim}80^{\circ}C$ for soluble TG and immobilized TG, respectively. The thermal stability of immobilized TG was significantly improved than that of soluble TG, and immobilized TG retained $32{\sim}40%$ higher activity than soluble TG. D-values from thermal inactivation of immobilized TG were 7690 sec at $65^{\circ}C$, 83 sec at $75^{\circ}C$, 7.2 sec at $80^{\circ}C$. Z-values of soluble and immobilized TG were $6.4^{\circ}C\;and\;5.3^{\circ}C$, respectively. The little difference of activation energies of soluble TG and immobilized TG supposed that there was little difference in mass transfer limitation during the reaction of soluble TG and immobilized TG.

• Journal of the Korean Society of Food Science and Nutrition
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• 제17권3호
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• pp.198-204
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• 1988

This study was aimed at obtaining elementary data on enzymatic browning of potato and potato products and examining the inhibitory method of browning. Therefore, we extracted polyphenol oxidase from potatoes(Solanum tubersum L.), and investigates its general properties and inhibiting effects of its activity with the different concentrations of sulfites($Na_2S_2O_4,\;Na_2SO_3{\cdot}7H_2O,\;NaHSO_3$). The optimum pH and temperature of polyphenol oxidase were observed to be 6.5 and $37^{\circ}C$ respectively. The polyphenol oxidase at PH5 was very stable, and the activity of polyphenol oxidase between pH $5.0{\sim}9.0$ was estimated to be relatively high, showing $72{\sim}75%$ of its activity at pH5. The polyphenol oxidase was very stable when heated at $40^{\circ}C$ for one hour, and almost 50% of enzyme activity was decreased when heated at $70^{\circ}C$ for twelve minutes. At 0.1mM concentrating of sulfites the relative activity of polyphenol oxidase was 98% in all the three cases of sulfites. Thus sulfites at 0.1mM concentration was found to have little inhibiting effect on polyphenol oxidase activity. At 1mM concentration of sulfites $NaHSO_3$ showed the lowest 36% relative activity among the three. At 5mM concentration of sulfites, the relative activity of $Na_2SO_3{\cdot}7H_2O$ was the lowest 14%.

• Korean Journal of Food Science and Technology
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• 제53권4호
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• pp.382-390
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• 2021

In this study, solubilized yuzu pulp powder (EYP) was produced using enzyme treated yuzu pulp powder (YP) and used to manufacture yuzu granules (0-20% EYP content). The physicochemical, product stability, and functional properties of Yuzu granules were compared among five enzyme treatments. Among the five treatments, CL had the highest YP solubilization yield (48.68%). Microstructural observation of EYP using FE-SEM revealed that its surface became irregular and porous after enzymatic treatment. Compared to YP, EYP had 2 times lower insoluble dietary fibers and 3 times lower hemicellulose and cellulose content. Among the yuzu granules, IV (yuzu granules with 15% EYP) had an excellent water and oil holding capacity and flowability. IV granule had the highest narirutin and hesperidin content of 3.4 mg and 2.2 mg/g DW, respectively and the highest antioxidant (68.4%) and tyrosinase inhibitory activities (82.5%). Therefore, EYP or granule with EYP can be used as a functional component in food industry or pharmaceutical field.

• Korean Journal of Agricultural Science
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• 제39권3호
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• pp.395-405
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• 2012

Lipozyme TL IM-catalyzed esterification was carried out to produce functional hard fat (structured lipid, SL) using palm stearin (PS) and hydrolysate of pomegranate seed oil (HPSO) of 1:6 molar ratio. HPSO contained conjugated linolenic acid (CLnA, about 80%). The reaction was performed at non-solvent system and solvent (n-hexane) system using Lipozyme TL IM (10% of total substrates, w/w) for 12, 24, and 72 hr in a shaking water bath ($55^{\circ}C$ and 185 rpm), respectively. SL synthesized in non-solvent system (NH-SL) and SL synthesized in n-hexane system (H-SL) were refined after deacidification, respectively. Their physicochemical properties were compared to obtain desirable functional hard fat. The content of CLnA in NH-SL increased from 34.38% to 40.63% with increasing reaction time. Similar results also observed in H-SL resulting in 36.81~45.83% of CLnA. In triacylglycerol (TAG) composition, the main molecules of LnLnLn (Ln=linolenic acid, PN=36) and the LnLnP (P=palmitic acid, PN=40) were newly synthesized in NH-SL and H-SL with increasing reaction time. After 72 hr reaction, iodine values of NH-SL (136.49) and H-SL (140.37) showed high values because of the high content of CLnA. Solid fat index (SFI) in NH-SL was higher than that in H-SL at each measured temperature. The predominant polymorphic forms of NH-SL and H-SL obtained after esterification for 72 hr were the desirable crystalline structure of the ${\beta}$' form.

• Korean Journal of Agricultural Science
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• 제12권2호
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• pp.324-332
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• 1985

Aspergillus niger B-15 with strong Endo-polygalacturonase (Endo-PG) activities was selected out from a total of 1,573 fungal strains isolated from various testing materials. A mutant strain, U-46, was obtained from the Aspergillus niger B-15 by repeated irradition of ultra-violet light. The objectives of the study were to investigate the fungal properties of the parental and mutant strains obtained and to study the condition of enzyme production and reaction. The results obtained are summarized as follows: 1. The size of conidial head of the U-46 mutant was smaller than that of the parental strains, B-15 and the length of the conidiophore was also shorter than that of the parental strains. 2. The optimum conditions for the Endo-PG production of the parental B-15 strain in the wheat bran Koji were obtained when 40% of water was added to the wheat bran and the temperature was 30 to $35^{\circ}C$. However, the best condition for the mutant U-46 strain was attained when 60 to 70% of water was added and the temperature was $35^{\circ}C$. The optimum growing periods were two to three days for both parental and mutant strains. 3. Under the optimum producing conditions of each strains, the enzymatic activity of the mutant U-46 was 20 times higher than the Endo-PG of the parental strain, B-15. 4. When both strains were cultured in the wheat bran Koji containing 60% of water at $35^{\circ}C$ for three days, the mutant strain. U-46, was about 46 times higher in the Endo-PG activity and about 18 times greater in Exo-PG activity than the parental strain, B-15. The activities of cellulase, $\alpha$-amylase, and glucoamylase were also highly increased in the mutant strain. 5. The mutant strain, U-46, increased its Endo-PG activity up to 20% over that of ordinary case when 1.2 to 1.5% of ammonium sulphate was added to the wheat bran. 6. The optimum condition for Endo-PG activity of crude enzyme of the mutant strain, U-46, was attained when pH of reaction solution was 4.0 to 4.5 and the temperature was $50^{\circ}C$.

• Korean Chemical Engineering Research
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• 제51권1호
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• pp.111-115
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• 2013

The optimum synthetic conditions of poly(p-phenylphenol) by horseradish peroxidase in dioxane:water (80:20 v/v) mixtures were studied. The stability against thermal degradation and structural properties of the synthesized phenolic resins were investigated by thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC), respectively. The synthetic yield of poly(p-phenylphenol) increased upon the increase of the amount of enzyme up to 0.25 mg HRP/mL, then leveled off for further increase of the enzyme usage. When sodium acetate (100 mM, pH 4~6) and sodium phosphate (100 mM, pH 7~9) were used as the buffering salts for the aqueous component (20% v/v), the synthetic yield of the resin increased at higher pH of the aqueous buffer. But when the pHs of the aqueous buffer were 6 and 9, the synthetic yield strongly depended on the types of the buffering salts; if sodium phosphate was used instead of sodium acetate at pH 6, the yield decreased by about 15% and if sodium bicarbonate was used instead of sodium phosphate, the yield decreased by almost 20%. When the pH range of the aqueous buffer was from 4 to 7, the addition of a radical mediator, 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonate) (ABTS), up to 2 mM improved the synthetic yield of the resin by about 10%. TGA experiments revealed that the thermal stability of the resin synthesized in dioxane:water (100 mM sodium phosphate, pH 9) (80:20 v/v) was high having the char yield of 47% upon the heating at $800^{\circ}C$. DCS results showed that the structures of the polymers synthesized in acidic aqueous buffers were different from those of the polymers synthesized in the basic aqueous buffers. However, all the synthesized resins were found to have the property of the thermosetting resins.

• Korean Journal of Agricultural Science
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• 제11권2호
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• pp.223-232
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• 1984

The influences of some factors involved in removing glucose from egg white by the glucose oxidase system be fore drying were investigated. And the properties between foams prepared from raw and enzyme-treat ed egg white was compared. The results obtained we re summarized as follows; 1. The dianisidine method was found to be suitable for the measurement of egg white glucose in the range up to 100ug/ml. 2. The optimal pH of glucose oxidase activity on glucose was found to be a bout 5.0, and thats activity was most stable in the pH range of about 4.0~5.0 when that enzyme was treat ed for 30 minute at $50^{\circ}C$. 3. The optimal temperature for glucose oxidase reaction on glucose was found to be about $20^{\circ}C$, and that enzyme activity was s table up to $50^{\circ}C$. 4. The removing rate of glucose from egg white with glucose oxidase was influenced by the enzyme concentration, pH and oxygen addition, and the react ion time of the desugarization was about 10 hour sunder the conditions of 0.5% hydrogen peroxide, pH 7.0 and $26^{\circ}C$. 5. All of the each egg white treated with glucose oxidase, glucose oxidase+pancreatin, glucose oxidase+trypsin showed highly foaming ability than that of natural egg white(control), but thats foam stability, on the contrary, was reversed.

• Korean Journal of Microbiology
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• 제51권2호
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• pp.97-107
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• 2015

Wetlands constitute a transitional zone between terrestrial and aquatic ecosystems and have unique characteristics such as frequent inundation, inflow of nutrients from terrestrial ecosystems, presence of plants adapted to grow in water, and soil that is occasionally oxygen deficient due to saturation. These characteristics and the presence of vegetation determine physical and chemical properties that affect decomposition rates of organic matter (OM). Decomposition of OM is associated with activities of various extracellular enzymes (EE) produced by bacteria and fungi. Extracellular enzymes convert macromolecules to simple compounds such as labile organic carbon (C), nitrogen (N), phosphorus (P), and sulfur (S) that can be easily taken up by microbes and plants. Therefore, the enzymatic approach is helpful to understand the decomposition rates of OM and nutrient cycling in wetland soils. This paper reviews the physical and biogeochemical factors that regulate extracellular enzyme activities (EEa) in wetland soils, including those of ${\beta}$-glucosidase, ${\beta}$-N-acetylglucosaminidase, phosphatase, arylsulfatase, and phenol oxidase that decompose organic matter and release C, N, P, and S nutrients for microbial and plant growths. Effects of pH, water table, and particle size of OM on EEa were not significantly different among sites, whereas the influence of temperature on EEa varied depending on microbial acclimation to extreme temperatures. Addition of C, N, or P affected EEa differently depending on the nutrient state, C:N ratio, limiting factors, and types of enzymes of wetland soils. Substrate quality influenced EEa more significantly than did other factors. Also, drainage of wetland and increased temperature due to global climate change can stimulate phenol oxidase activity, and anthropogenic N deposition can enhance the hydrolytic EEa; these effects increase OM decomposition rates and emissions of $CO_2$ and $CH_4$ from wetland systems. The researches on the relationship between microbial structures and EE functions, and environmental factors controlling EEa can be helpful to manipulate wetland ecosystems for treating pollutants and to monitor wetland ecosystem services.

• Journal of Life Science
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• 제26권10호
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• pp.1105-1112
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• 2016

The aim of this study was to examine the metabolic adjustment of lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes to the environmental temperature in bluegill (Lepomis macrochirus). This study included three groups of bluegill collected in April (group Ⅰ), May (group Ⅱ), and September (group Ⅲ). The LDH activities of skeletal muscle, heart, and brain tissues were higher in group Ⅲ than in groups Ⅰ and Ⅱ. The citrate synthase (EC 4.1.3.7, CS) activity was higher in skeletal muscle but lower in heart and brain tissues of group Ⅱ as compared to group Ⅰ. In contrast, the CS activity was lower in skeletal muscle and higher in heart and brain tissues in group Ⅲ than in group Ⅱ. Furthermore, the LDH/CS activity ratio was higher in the skeletal muscle and brain in group Ⅲ than in groups Ⅰ and Ⅱ. Accordingly, anaerobic metabolism was increased in group Ⅲ. LDH A₄, A₂B₂, and B₄ isozymes were expressed in skeletal muscle, heart, liver, and brain tissues. The LDH C hybrid was detected in brain tissue. The LDH A₄ isozyme was successfully purified by affinity chromatography. The molecular weight of the purified LDH A₄ isozyme was 136 kDa and its optimal pH for enzymatic activity was 8.0. The K_m^PYR values of LDH in skeletal muscle were 0.161-0.227 mM using pyruvate as a substrate. These kinetic properties of LDH in skeletal muscle are consistent with the fact that bluegill is a cold-adapted species. These results may be useful for predicting the habitat use of this fish.