• Journal of Environmental Science International
• /
• v.11 no.12
• /
• pp.1315-1320
• /
• 2002

Since the enzymatic degradation of microbial poly[(R)-3-hydroxybutyrate-co-3-hydroxyvalerate] (P(3HB-co-3HV)) initially occurs by a surface erosion process, a degradation behavior could be controlled by the change of surface property. In order to control the rate of enzymatic degradation, plasma gas discharge and blending techniques were used to modify the surface of microbial P(3HB-co-3HV). The surface hydrophobic property of P(3HB-co-3HV) film was introduced by CF$_3$H plasma exposure. Also, the addition of small amount of polystyrene as a non-degradable polymer with lower surface energy to P(3HB-co-3HV) has been studied. The enzymatic degradation was carried out at 37 $^{\circ}C$ in 0.1 M potassium phosphate buffer (pH 7.4) in the presence of an extracellular PHB depolymerase purified from Alcaligenes facalis T1. Both results showed the significant retardation of enzymatic erosion due to the hydrophobicity and the enzyme inactivity of the fluorinated- and PS-enriched surface layers.

• Journal of Microbiology and Biotechnology
• /
• v.4 no.2
• /
• pp.113-118
• /
• 1994

A bacterial strain NS-83 isolated from soil was able to produce an extracellular thermostable protease. The strain was identified as Pseudomonas aeruginosa based on its morphological and physiological characteristics. A thermostable protease from this strain has been purified to homogeneity as judged by SDS-PAGE and isoelectric focusing. The purification procedures included hydrophobic interaction, ion exchange, and gel filtration chromatography. The $M_r$ and the pl of the enzyme were 32,000 and 5.9, respectively. The optimal pH at 55$^{\circ}C$ and the optimal temperature at pH 7.0 were 8.0 and 60$^{\circ}C$, respectively. The D-values of the enzyme at 60, 65, and 70$^{\circ}C$ were 22, 2.1, and 0.75 hrs, respectively. The enzyme activity was significantly inhibited in the presence of 1 mM o-phenanthroline or EDTA, suggesting that the enzyme is metalloprotease. The $K_m$, and $V_{max}$ for Hammarsten casein were found to be 3.2 mg/ml and 0.918 unit/ml, respectively. These enzymatic properties were similar to those of elastase produced from P. aeruginosa IFO 3455, but the enzyme was clearly different from the reported elastase, in respect to $Ca^{++}$ effects on enzyme-thermostability. This property, together with amino acid composition analysis, confirmed that the enzyme differs from the known P. aeruginosa elastase.

• The Korean Journal of Food And Nutrition
• /
• v.36 no.1
• /
• pp.42-49
• /
• 2023

To enhance the efficacy of Abeliophyllum distichum leaves, extracts were prepared using different solvents for hydrolytic enzyme-treated Abeliophyllum distichum leaves. Physicochemical quality and antioxidant activity were measured. Soluble solids, reducing sugar, ascorbic acid, flavonoids, and polyphenols contents showed the lowest values in the control without enzyme treatment. However, they showed high contents in ethanol extract. In the case of enzyme treatment, their values were higher than those of the control. In particular, verbascoside content increased about 220 times more than that of the control group when treated with enzymes and extracted with 50% ethanol. pH was lowered upon enzymatic treatment. Regarding DPPH radical scavenging activity, for enzyme-free, 25% ethanol extract showed the highest activity among extracts with different solvents. For cellulase and pectinase-treated leaves, water extract showed the highest DPPH radical scavenging activity among extracts with different solvents. For leaves treated with enzyme combination, 50% ethanol extract showed the highest DPPH radical scavenging activity among extracts with different solvents. Regarding ABTS radical scavenging activity, it was generally higher in the 50% ethanol extract than in the water extract and 25% ethanol extract. In particular, verbascoside content was increased when the extract was prepared by co-treatment with enzymes and 50% ethanol.

• Journal of the Korean Institute of Electrical and Electronic Material Engineers
• /
• v.15 no.7
• /
• pp.621-626
• /
• 2002

In the case of immobilizing of glucose oxidase (GOx) in polypyrrole (PPy) conducting polymer using electrosynthesis, the GOx obstructs charge transfer and mass transport during the film growth. This may lead to short chained polymer and/or make charge-coupling weak between the GOx and the backbone of the PPy. That is mainly due to insulating property and net chain of the GOx. Since being the case, it is useless to increase in amount of GOx mere than reasonable in the synthetic solution. We improved the amount of immobilized GOx into the PPy by adding a little ethanol in the synthetic solution without any more amount of GOx in the solution. We electrochemically analyzed an improvement in the immobilizing event. For the glucose sensing, when ethanol was added by 0.1 mol $dm^{-3}$ in the synthetic solution, the Michaelis constant of the resulting enzyme electrode was about 32 mmol $dm^{-3}$ and maximum current was about $146\mu A$.

• Journal of Life Science
• /
• v.9 no.2
• /
• pp.44-48
• /
• 1999

The ionization of tyrosine residue is known to be involved in the stabilization of transition-state in catalysis of astacin based upon the astacin-transition state analog structure. Two tyrosine residues, Tyr-216 and Tyr-219, are conserved in all MMPs related with astacin family, We replaced Tyr-216 and Tyr-219 into phenylalanine, respectively and the zinc binding properties, kinetic parameters, and pH dependence of each mutant are determined in order to examine the role of tyrosine residue in matrilysin catalysis. Both mutants contain two zinc atoms per mol of enzyme, indicating that either tyrosime does not affect the zinc binding property of the enzyme. Y216F and Y219F mutants are highly active and the kcat/Km values are only decreased 1.1-1.5-fold compared to the wild-type enzyme. The decrease in the activity of the mutants is essentially due to the increase in Km value. The pH dependencies of the kcat/Km values for both mutants are similar to the corresponding dependencies obtained with the wild type enzyme. The pKa values at the alkaline side of both mutants are not changed. These kinetic and pH dependence results indicate that the ionization of active site tyrosine residue of matrilysin is not reflected in the kinetics of peptide hydrolysin as catalyzed by astacin.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.31 no.3
• /
• pp.465-468
• /
• 2002

The galactose/mannose ratio of guar gum, guar gum treated with purified ${\alpha}$-galactosidase and locust bean gum were investigated. Gel-promoting property of enzyme-treated guar gum increased when the galactose/mannose ratio was about 1 : 3.2, which was close to the ratio of 1 : 3.3 for locust bean gum. And the ratio was obtained when the guar gum was hydrolyzed by the enzyme for 24 hr. It is clear that enzymatic depletion of galactose from guar gum by sunflower seed ${\alpha}$-galactosidase would lead to a significant increase in gelation ability. The mixture of xanthan gum and guar gum, and xanthan gum, guar gum and enzyme-treated copra meal were also investigated in viscosity behavior.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.26 no.6
• /
• pp.1116-1121
• /
• 1997

Effects of gamma irradiation onf the activity and the properties(amino acid compositions, in vitro digestibility and SDS-PAGE pattern) of proteolytic enzymes were investigated. The proteolytic activity of soluble human serine protease, enzyme in kiwi and pineapple decreased 10% and 30~65% by 5 kGy and 30 kGy, respectively. In dried pancreatin and lysozyme, the proteolytic and antimicrobial activities decreased 6~14% and 10~20% by 5kGy and 40kGy, respectively. The analysis of above 10kGy-irradiated soluble human serine protease by SDS-PAGE revealed radiolysis of the enzyme into protein or peptides of lower molecular weights. The irradiation of skim milk, hammastein casein, and lysozyme up to 40kGy had no deleterious effect on either the in vitro digestibility or amino acid compositions.

• Applied Chemistry for Engineering
• /
• v.10 no.5
• /
• pp.635-638
• /
• 1999

The effect of proteolytic enzyme on the unhairing process has been studied. this enzyme significantly changed the physical properties of pelt for leather shoes. A SEM was used to examine physical properties of the pelt. Even though bio-tech treatment using proteolytic enzyme slightly reduced the degree of unhairing compared to chemical treatment. Physical property of the pelt is better and this method is environmentally favorable. $H_{2}S$ gas produced in the chemical treatment was not detected and the value of COD and BOD for waste water after the unhairing process were reduced to 939 mg/L and 5268 mg/L, respectively. We found that a process with 0.4~0.5% proteolytic enzyme for 20 h at $29{\sim}30^{\circ}C$ is most suitable for the unhairing process.

• BMB Reports
• /
• v.29 no.2
• /
• pp.111-115
• /
• 1996

A novel glutathione S-transferase from Pseudomonas sp. DJ77 was expressed in E. coli and purified by glutathione-affinity chromatography. The enzyme was composed of two identical subunits. The molecular size of the enzyme was 42 kDa by sephadex G-150 gel permeation chromatography and Mr of each subunit was 23 kDa by sodium dodecylsulfate-polyacrylamide gel electrophoresis. pI value of the enzyme was approximately 5.8 by isoelectric focusing. This enzyme showed the highest activity toward 1-chloro-2,4-dinitrobenzene as the electrophilic substrate. The relative activities toward p-nitrobenzyl chloride and 1,2-dichloro-4-nitrobenzene were 3.8% and 1.3% of the activity toward 1-chloro-2,4-dinitrobenzene, respectively. $K_m$ and $V_{max}$ values for 1-chloro-2,4-dinitrobenzene calculated by Lineweaver-Burk plot were 0.76 mM and $14.81\;{\mu}mol/min/mg$, respectively, and those for glutathione were 6.23 mM and $64.93\;{\mu}mol/min/mg$, respectively. The enzyme showed highest glutathione S-transferase activity at pH 8.0 and was stable between pH 6.0 and 9.0. The enzyme retained its activity up to $35^{\circ}C$ for 90 min but was unstable above $45^{\circ}C$.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.25 no.4
• /
• pp.687-694
• /
• 1996

Myrosinase was purified from Korean mustard seed(Brassica juncea) by a sequential process of DEAE-cellulose, concanavalin A-sepharose, and Superose 6 chromatography. The molecular weight of puri-fied myrosinase(II-2) determined by SDS-polyacrylamide electrophoresis was 67KD. About a 248-fold purification for myrosinase II-2 was obtained after Superose 6 chromatography. Optimum pH of the myrosinase was 7.0 and optimum temperature of the enzyme was $3^{\circ}C.$ The enzyme was stable at pH 7.0, and below $30^{\circ}C.$ Cu, Hg and Fe ion significantly inhibited the enzyme activity, but ascorbic acid enhanced, resulting in a maximum activity by 1mM ascorbic acid. Among tile ascorbic acid ana-logues, dehydroascorbic acid inhibited the enzyme activity, whereas others showed a little effect. Reducing agents such as 2-mercaptoethanol and dithiothreitol inhibited the enzyme activity, but the reducing agents with ascorbic acid was enhanced enzyme activity.