• Microbiology and Biotechnology Letters
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• v.22 no.6
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• pp.636-642
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• 1994

Essential amino acids involving in the catalytic mechanism of the $\beta$-D-xylosidase of Bacillus stearothermophilus were determined by chemical modification studies. Among various che- mical modifiers tested N-bromosuccinimide (NBS), $\rho$-hydroxymercurybenzoate (PHMB), N-ethylma- leimide, 1-[3-(di-ethylamino)-propyl]$-3-ethylcarbodi-imide (EDC), and Woodward's Reagent K(WRK)inactivated the enzyme, resulting in the residual activity of less than 20%. WRK reduced the enzyme activity by modifying carboxylic amino acids, and the inactivation reacion proceeded in the form of pseudo-first-order kinetics. The double-lagarithmic plot of the observed pseudo-first- order rate constant against the modifier concentration yielded a reaction order of 2, indicating that two carboxylic amino acids were essential for the enzyme activity. The $\beta$-D-xylosidase was also inactivated by N-ethylmaleimide which specifically modified a cysteine residue with a reaction order of 1, implying that one cysteine residue was important for the enzyme activity. Xylobiose protected the enzyme against inactivation by WRK and N-ethylmaleimide, revealing that carboxylic amino acids and a cysteine residue were present at the substrate-binding site of the enzyme molecule.

• Journal of Dairy Science and Biotechnology
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• v.26 no.2
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• pp.39-43
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• 2008

EMC have a similar enzymatic reaction to cheese, but the EMC produce the stronger flavors than cheese by much more enzymatic reaction. It is important to find appropriate enzyme in order to develop these kind of superior EMC. Calf PGE is more suitable than that of kid and lamb to develop the mild cheese flavors. Especially, it was known that animal esterase and peptidase were more benefit than microbial enzyme for Cheddar cheese flavors. On the Cheddar and Swiss cheese, EMC flavors were much more 3 times than the cheese flavors. In the ratio of each component, butyric acid, myristic acid, palmitic acid and oleic acid were high in free fatty acid, and glutamic acid, valine, leucine and lysine were high in free amino acid of the Cheddar EMC.

• Clean Technology
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• v.22 no.3
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• pp.175-180
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• 2016

Tyrosinases catalyze the hydroxylation of a monophenol (monophenolase activity) and the conversion of an o-diphenol to o-quinone (diphenolase activity), which are mainly involved in the modification of tyrosine residues into 3,4-dihydroxyphenyl-alanine (DOPA) and DOPA/DOPAquinone-derived intermolecular cross-linking. Previously, we obtained a slightly acidic and cold-active tyrosinase, tyrosinase-CNK, by our recombinant protein approach. The enzyme showed optimal activity at pH 6.0 and 20 ℃ with an abnormally high monophenolase/diphenolase activity ratio and still had approximately 50% activity compared with the highest activity even in ice water. Here, we investigated reaction stability of the recombinant tyrosinase-CNK as a psychrophilic enzyme. The enzyme showed remarkable thermal stability at 0 ℃ and the activity was well conserved in repeated freeze-thaw cycles. Although water-miscible organic solvent as reaction media caused the activity decrease of tyrosinase-CNK as expected, the enzyme activity was not additionally decreased with increased concentration in organic solvents such as ethanol and acetonitrile. Also, the enzyme showed high salt tolerance in chaotropic salts. It was remarkably considered that 2⁺ metal ions might inhibit the incorporation of Cu²⁺ into the active site. We expect that these results could be used to design tyrosinase-mediated enzymatic reaction at low temperature for the production of catechols through minimizing unwanted self-oxidation and enzyme inactivation.

• Microbiology and Biotechnology Letters
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• v.10 no.1
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• pp.45-52
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• 1982

A fungus with the highest protopectinase productivity was selected among 205 strains isolated from the soil and identified as a Verticillium sp. The Verticillium sp. was cultivated on wheat bran and the crude extruct of its culture medium showed the highest protopectinase activity on the following conditions: 3 days of cultivation time, 27$^{\circ}C$ of cultivation temperature, 1.2 $m\ell$/g wheat bran of water content, and reinforcement of ammonium nitrate and calcium chloride at the concentration of 0.5 and 0.02%, respectively. The optimum conditions for pectin production from Citrus peel pulp by the protopectinase were consequently obtained as follows: 20$m\ell$/g of liquid volume-to-pulp weight ratio, 4$0^{\circ}C$ of reaction temperature, and 4 of reaction pH. The higher the enzyme concentration, the better the yield of pectin and the shorter the reaction time. Total 45.6mg of pectin/g peel was produced by 1 hour reaction at the enzyme concentration of 10.5 units/$m\ell$. Molecular weight of the pectin produced by the enzyme was estimated to be about 62,000 by Smit and Bryant's method.

• Journal of Microbiology and Biotechnology
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• v.19 no.12
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• pp.1596-1602
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• 2009

A lipase-catalyzed esterification reaction of (S)-naproxen ethyl ester by CALB (Candida antarctica lipase B) enzyme was performed in supercritical carbon dioxide. Experiments were performed in a high-pressure cell for 10 h at a stirring rate of 150 rpm over a temperature range of 313.15 to 333.15 K and a pressure range of 50 to 175 bar. The productivity of (S)-naproxen ethyl ester was compared with the result in ambient condition. The total reaction time and conversion yields of the catalyzed reaction in supercritical carbon dioxide were compared with those at ambient temperature and pressure. The experimental results show that the conversion and reaction rate were significantly improved at critical condition. The maximum conversion yield was 9.9% (216 h) at ambient condition and 68.9% (3 h) in supercritical state. The effects of varying amounts of enzyme and water were also examined and the optimum condition was found (7 g of enzyme and 2% water content).

• Korean Journal of Veterinary Research
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• v.32 no.3
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• pp.429-434
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• 1992

This study was carried out to develop an effective liquid-phase double antibody enzyme immunoassay for determining of progesterone. The optimum conditions of assay system, 1st and 2nd antibodies, enzyme conjugate, and time reaction were invested. The bovine plasma progesterone level in dairy cattle and korean native bulls were also analyzed. The results obtained were as follows; 1. The reproducibility of petroleum ether was superior to that of ethyl ether as extract solvent of progesterone in plasma. 2. The optimum dilution rate of 1st and 2nd antibody was 30,000 and 10 times, respectively. Affer the reaction of enzyme conjugate to progesterone 1st antibody, and then 2nd antibody competition reaction was enough for over 1hr. 3. Average plasma progesterone level in 4 pregnant and 9 nonpregnant Holstein was $2.5{\pm}0.5$ and $0.7{\pm}0.2ng/m{\ell}$, respectively. Average plasma progesterone level of 10 Korean native bulls was $0.1{\pm}0.001ng/m{\ell}$ From these results, by using liquid phase double antibody enzyme immunoassay for progesterone is applicable to detect of early pregnancy diagnosis, factorial analysis of reproductive disorder, and also reproductive physiological function such as monitoring of cyclicity during the post-partum period.

• Korean Journal of Food Science and Technology
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• v.10 no.2
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• pp.209-214
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• 1978

Naringinase produced from Aspergillus nidulans was immobilized in acrylamide gel by the entrapping method and its characteristics were studied. Optimum acrylamide concentration was 10%, but N.N'-methylene bisacrylamide concentration had no influence on the final enzyme gel activity. The suitable amount of enzyme dissolved in the polymerization reaction mixture was 126 units/ml. Optimum pH of immobilized enzyme was 5.0 which was the same as that of free enzyme. However, immobilized enzyme showed a higher optumum reaction temperature, markedly increased pH and temperature stability. In a packed-column reactor, the observed reaction rate was increased proportionally to flow rate up to 5ml/min., but independent above 6ml/min.. Activation energy of the immobilized enzyme was 13.01 Kcal/mole, and the energy required for the thermal inactivation was 39.4 Kcal/mole. The apparent Km for 100 mesh gel was $7.23{\times}10^{-3}$ mole.

• Korean Journal of Animal Reproduction
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• v.24 no.4
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• pp.339-341
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• 2000

In mammals, fatty acid utilization is initiated by activation of fatty acid, catalyzed by acyl-CoA synthetase(ACS, EC6.2.1.3). This enzyme reaction is essential in fatty acid metabolism, since mammalian fatty acid synthetase contains a specific thioesterase to produce fatty acid as th $\varepsilon$ final reaction product. Acyl-CoA, the product of ACS, is utilized in various metabolic pathways including membrane biogenesis, energy production and fat deposition. (omitted)

• The Korean Journal of Food And Nutrition
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• v.30 no.6
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• pp.1359-1363
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• 2017

The purpose of this study was that the optimal hydrolysis conditions of endo- and exo-type enzymes were selected to utilize organic cheese byproducts. Optimal substrate concentration and optimum enzyme ratio were measured by using 4 kinds of endo-type enzymes (alcalase, neutrase, protamex, and foodpro alkaline protease) and two exo-type enzymes (flavourzyme and prozyme 2000P) for whey protein hydrolysis were analyzed using liquid chromatography. As a result, the optimal endo-type enzyme through the first enzyme reaction was selected as alcalse, and as a result of the secondary enzyme reaction, flavourzme was selected as the Exo type enzyme. The concentration of whey protein substrate for optimal primary and secondary enzyme reactions was 10%. In addition, the optimum ratio of enzyme was 0.5% of alcalase and 0.2% of flavourzyme, which showed low molecular weight chromatography pattern compared to 2% of alcalase and 1% of flavourzyme hydrolyzate. Therefore, hydrolyzing the endo-type enzyme alcalase at a concentration of 0.5% for 10 hours and then hydrolyzing the exo-type enzyme flavouryme at a concentration of 0.2% for 4 hours was considered to be the optimum condition.

• Microbiology and Biotechnology Letters
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• v.10 no.1
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• pp.1-7
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• 1982

A Rhizopus sp. was selected for its strong cellulolytic activity among various strains of molds found in rotting ginseng roots. Studies were made on some properties of the cellyloiytic enzyme produced by the strain. The results obtained were summarized as follows: The optimum pH of the enzyme was 4.5 and the range of its stability to the pH was 3.0 to 7.0. The optimum temperature was 5$0^{\circ}C$, while the enzyme was instantly inactivated above 6$0^{\circ}C$. Mn$^{＋＋}$ and Co$^{＋＋}$ ions increased enzyme activity and the metal ions were found to increased the ther-mostability of the enzyme. This enzyme was inhibited by sodium dodecyl sulfate and 2,4-dinitrophenol. This enzyme had a strong cellulolytic enzyme activity on various native cellulose given a sufficient reaction time. The addition of 0.5% saponin solution into reaction mixture increased the enzyme activity.