• KSBB Journal
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• v.19 no.1
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• pp.50-56
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• 2004

The aim of this work was to investigate the purification and characterization dixoygenases isolated from Delftia sp. JK-2, which could utilize aniline, benzoate, and p-hydroxybenoate as sole carbon and energy source. Catechol 1,2-dioxygenase (C1, 2O), catechol 2,3-dioxygenase(C2, 3O), and protocatechuate 4,5-dioxygenase(4,5-PCD) were isolated by benzoate, aniline, and p-hydroxybenzoate. In initial experiments, several characteristics of C1 ,2O, C2, 3O, and 4,5-PCD separated with ammonium sulfate precipitation, DEAE-sepharose, and Q-sepharose were investigated. Specific activity of C1 ,2O, C2, 3O, and 4,5-PCD were approximately 3.3 unit/mg, 4.7 unit/mg, and 2.0 unit/mg. C1 ,2O and C2, 3O demonstrated their enzyme activities to other substrates, catechol and 4-methylcatechol. 4,5-PCD showed the specific activity to the only substrate, protocatechuate, but the substrates(e.g., catechol, 3-methylcatechol, 4-methylcatechol, 4-chlorocatechol, 4-nitrocatechol) did not show any specific activities in this work. The optimum temperature of C1, 2O, C2, 3O, and 4,5-PCD were 30$^{\circ}C$, and the optimal pHs were approximately 8, 8, and 7, respectively. Ag$\^$+/, Hg$\^$+/, Cu$\^$2+/ showed inhibitory effect on the activity of C1, 2O and C2, 3O, but Ag$\^$+/, Hg$\^$+/, Cu$\^$2+/, Fe$\^$3+/ showed inhibitory effect on the activity of 4,5-PCD. Molecular weight of the C1, 2O, C2, 3O, and 4,5-PCD were determined to approximately 60 kDa,35 kDa, and 62 kDa by SDS-PAGE.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.8
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• pp.1025-1030
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• 2006

In other to develop new functional doenjangs, two types of the isolated nuruk doenjangs were prepared with protease and amylase-producing Aspergillus oryzae D-2 and antihyperlipidemia Bacillus subtilis LK-12 and then changes of its quality and physiological functionalities were investigated during 2 months of fermentation and compared with those of the commercial nuruk doenjangs made by commercial Aspergillus oryzae and antihyperlipidemia Bacillus subtilis LK-12. ${\alpha}-Amylase$ activity of the isolated nuruk doenjangs during fermentation were decreased slightly, whereas proteases activities were increased significantly to $1.8{\sim}2.8$ Unit per mL after 1 month of fermentation. These ${\alpha}-amylase$ activities and proteases activities were similar with those of the commercial nuruk doenjangs. Amino-nitrogen content and reducing sugar content of the doenjangs after 2 months of fermentation were approximate $1.63{\sim}1.72\;mg%$ and $0.77{\sim}0.81%$, respectively. Antihypertensive angiotensin-Ⅰ converting enzyme inhibitory activities of the isolated nuruk doenjangs were slightly decreased from $85.6{\sim}87.2%$ to $84.0{\sim}85.1%$ after 2 months of fermentation and the commercial nuruk doenjangs were also significantly decreased from $85.7{\sim}88.0%$ to $69.1{\sim}79.7%$, lower than the isolated nuruk doenjangs. Fibrinolytic activity and HMG-CoA reductase inhibitory activity of the isolated nuruk doenjangs were very low and it were also similar with those of the commercial nuruk doenjangs. Antioxidant activity of the isolated nuruk doenjangs were showed $17{\sim}22%$, lower than that of the commercial nuruk doenjangs $(22{\sim}26%)$.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.8
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• pp.1086-1091
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• 2011

Anti-oxidative activity and tyrosinase inhibitory activity of various ethanol extracts of Polygoni multiflori radix (PMR) and Cynanchi wilfordii radix (CWR) were compared to identify an anti-oxidant and whitening agent source from nature. We conducted an investigation into the anti-oxidant activities of PMR and CWR ethanol extracts by measuring total polyphenol content, total flavonoid content, and ABTS radical capacity. The total polyphenol contents of PMR and CWR were 17.31${\pm}$0.54 mg GA/eq g, and 2.75${\pm}$0.22 mg GA/eq g, respectively. The total flavonoid contents of PMR and CWR were 6.38${\pm}$0.39 mg naringine/eq g, and 1.34${\pm}$0.09 mg naringine/eq g, respectively. The 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) radical decolorization of PMR and CWR were 96.89${\pm}$0.21% at 1 mg/mL and 93.49${\pm}$0.76% at 50 mg/mL. Melanoma cells were cultured with the PMR and CWR ethanol extracts for 48 hr, and total melanin content as a final product and the activity of tyrosinase, a key enzyme, in melanogenesis, were estimated. The PMR and CWR ethanol extracts increased melanin content and tyrosinase activity in a dose-dependent manner. These results suggest that PMR and CWR ethanol extracts could be useful as a skin whitening agent.

• Korean Journal of Food Science and Technology
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• v.39 no.3
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• pp.348-352
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• 2007

Inflammation is a complex process resulting from a variety of mechanisms. Combined inhibition of the activities of enzymes involved in the process may therefore be considered more important in anti-inflammatory property of plant extracts than any single contribution. In this study, the inhibitory effects of the ethanol extracts of thirty plant foods on the activities of secretory phospholipase $A_{2}$ ($sPLA_{2}$), cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), and 12-lipoxygenase (12-LOX) were examined. Several legumes, mungbean sprout and some leaf vegetables inhibited the activity of $sPLA_2$, upstream enzyme of inflammation pathway. Only soybean sprout and mungbean sprout significantly inhibited 12-LOX activity. Although most of extracts inhibited the activities of both COX-1 and COX-2, water dropwort and amaranth showed selectivity for the inhibition of COX-2 over COX-1. Especially, mungbean showed anti-inflammatory property at both upstream and downstream of inflammation pathway with relatively low $IC_{50}$ values for $sPLA_{2}$ and COX-2 enzymes. Mungbean sprout exhibited inhibitory effects on all enzymes related to early and late inflammation and soybean sprout suppressed 12-LOX and COX-2 simultaneously, although the activities of these plants were showed at relatively high concentration. Therefore, mungbean, mungbean sprout, and soybean sprout appear to exhibit anti-inflammatory effects by combined inhibition of inflammatory enzymes.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.8
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• pp.1087-1096
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• 2010

This study was conducted to investigate the physiological activity of extracts of fresh mushrooms. The components were extracted by hot water; subsequently, the hot-water extract was subjected to 60% ethanol precipitation to yield high-molecular-weight (HMW) and low-molecular-weight (LMW) fractions. Total polyphenol contents, $\beta$-glucan contents, electron-donating ability (EDA), superoxide dismutase (SOD)-like activity, nitrite-scavenging activity, fibrinolytic activity, nitric oxide (NO) production, and inhibition of NO production of the mushroom extracts were measured using lipopolysaccharide (LPS)-stimulated murine macrophages, RAW 264.7 cells. The extracts of Lentinus edodes (Berk.) Singer and Pleurotus ostreatus (Fr.) Kummer contained the highest levels of $\beta$-glucan (33.5% and 25.57%, respectively). Further, the LMW fractions of the Phellinus linteus contained the highest levels of polyphenols (233.23 mg/g). The EDA of LMW fractions (10 mg/mL) of the Phellinus linteus and Agaricus bisporus were 80.74% and 51.35%, respectively. Further, SOD-like activities of the LMW fractions were high as compared to those of the HMW fractions. Nitrite-scavenging activities of the LMW fractions (pH 1.2; concentration, 10 mg/mL) of the Phellinus linteus and Pleurotus ostreatus (Fr.) Kummer were 75.95% and 41.05%, respectively. The fibrinolytic activity of the LMW fractions of all mushrooms showed no enzyme activity by fibrin plate assay. The fibrinolytic activity of the extracts of Tricholoma matsutake was the greatest inhibitory activity at 60.4%. Further study revealed that the mushroom extracts exhibited anti-inflammatory effects on RAW 264.7 cells. The LMW fraction ($500\;{\mu}g/mL$) of the Phellinus linteus considerably inhibited NO production (100%).

• Journal of the Korean Society of Food Science and Nutrition
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• v.17 no.3
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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%.

• The Korea Journal of Herbology
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• v.22 no.3
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• pp.117-125
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• 2007

Objective : Lonicera japonica (Caprifoliaceae) has long been used for treatment of infectious diseases in oriental countries. The aim of this study was to investigative the effect by which the aqueous extract from flower of L. japonica (LJFAE) inhibited the lipopolysaccharide (LPS)-induced inflammatory mediators in murine macrophages, RAW 264.7 cells Methods : The dried flowers of L. japonica were extracted with distilled water at $100^{\circ}C$ for 7 h. The extract was filtered through 0.45 ${\mu}m$ filter, freeze-dried. The dried extract was dissolved in Hank's balanced salt solution (HBSS) and filtered through 0.22 ${\mu}m$ filter before use. Accumulated nitrite, an oxidative product of nitric oxide (NO), was measured in the culture medium by the Griess reaction. The levels of prostaglandin E2 (PGE2), tumor necrosis factor-$\alpha$ (TNF-$\alpha$), interleukin-1$\beta$ (IL-1$\beta$), and IL-6 production, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression were measured by enzyme-linked immunosorbent assay and Western blot analysis. Results: LJFAE (10-400 ${\mu}g$/ml) per se had no cytotoxic effect in unstimulated macrophages, but LJFAE concentration-dependently reduced NO, PGE2, TNF-, IL-l, and IL-6 production and COX-2 activity caused by stimulation of LPS. The levels of iNOS and COX-2 protein expressions were markedly suppressed by the treatment with LJFAE in a concentration dependent manner. Conclusions : These results suggest that LJFAE suppress the NO and PGE2production in macrophages by inhibiting iNOS and COX-2 expression and these properties may contribute to the anti-inflammatory activity of Lonicera japonica.

• KSBB Journal
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• v.11 no.2
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• pp.151-158
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• 1996

For the effective ethanol fermentation, the high concentration of sugar as the substrate of microbial fermentation is required. The most important reason in the inefficient hydrolysis; the easy deactivation of enzyme by temperature or shear stress and the severe inhibition effects of its products. In our work, we comprehended the kinetic characteristics of cellulose and ${\beta}$-glucosidase in the progress of hydrolysis, and observed the potential inhibitory effects of the hydrolyzed products and the deactivation of enzymes. We also tried to present the kinetic model of enzymatic hydrolysis of cellulose, which is applicable to process at the high concentration of sugar. Cellulase and ,${\beta}$-glucosidase exhibit diverse kinetic behaviors. At a level of only 5g/$\ell$ of glucose, the ${\beta}$-glucosidase activity was reduced by more than 70%. This result means that ${\beta}$-glucosldase was the most severely inhibited by glucose. Also at l0g/$\ell$ of cellobiose, the cellulose lost approximately 70% of its activity. ${\beta}$-glucosldase was more sensitive to deactivation than cellulose by about 1.6 times. The comprehensive kinetic model in the range of confidence was obtained and the agreement between the model prediction and the experimental data was reasonably good, testifying to the validity of the model equations used and the associated parameters.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.46 no.1
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• pp.37-45
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• 2013

This study was conducted to improve the yield of extracts from Alaska pollock Theragra chalcogramma head and sea tangle Laminaria japonica byproducts using various commercial enzymes, such as Alcalase, Flavourzyme, Neutrase (NH), and Protamex. Among the enzymatic hydrolysates, the yield was highest in hydrolysate incubated with NH for 4 h. NH-treated hydrolysates (NHH) also improved functional properties, such as angiotensin-I converting enzyme (ACE) inhibitory activity and 2,2-diphenyl-1-picryldrazyl (DPPH) radical scavenging activity, as compared to extracts from Alaska pollock head and sea tangle byproducts. Total free amino acid and taste values of NHH were 379.7 mg/100 mL and 24.03, respectively, after digestion for 4 h. These values are 2.2-fold and 1.9-fold higher compared with those of water soluble fractions extracted from Alaska pollock head and non-forming sea tangle, respectively. According to the taste value results, the major taste-active compounds among free amino acids of NHH were glutamic acid and aspartic acid. These results suggest that NHH can be used as an ingredient for natural seasoning preparation.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.33 no.1 s.60
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• pp.53-60
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• 2007

(6R)-5,6,7,8-tetrahydrobiopterin ($6-BH_4$) cofactor is essential for various process, and is present in probably every cell or tissue of higher organism. $6-BH_4$ is required lot various enzyme activities, and for less defined functions at the cellular level. And it is well known about the antioxidant effects as a non-protein compound. Recently, scientists proposed another roles for $6-BH_4$ in melanogenesis. $6-BH_4$ is a well known tyrosinase inhibitor. In this study, we found that methyl-$BH_4$ and $6-BH_4$ have antioxidant activities and inhibitory activity for melanin synthesis. These pterin compounds were not toxic in HaCaT and B16F10 cells and showed scavenging activity against DPPH radicals. We also showed that pterin compounds decreased protein levels of tyrosinase and TRP-1. In a clinical test, pterin compounds showed the significant skin whiteining effect after treatment for 3 weeks. Furthermore pterin compounds significantly suppressed the UVB-induced expression of $PGE_2$ and IL-6 genes induced UVB In HaCaT and inhibited UVB-induced melanogenesis in B16F10 cells. These results showed the effect of pterin compounds as a cosmeceutical ingredient.