• Microbiology and Biotechnology Letters
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• v.37 no.2
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• pp.99-104
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• 2009

The $\beta$-glucosidase gene from Streptomyces coelicolor A3(2) was cloned and expressed in Escherichia coli. The ORF consisted of 1377 nucleotides encoding 51 kDa in a predicted molecular weight. Effects of pH indicated that the $\beta$-glucosidase showed similar activity using $\alpha$-pNPG($\rho$-nitrophenyl-$\alpha$-D-glucopyranoside), $\beta$-pNPG($\rho$-nitrophenyl-$\beta$-D-glucopyranoside), and $\beta$-pNPF($\rho$-nitrophenyl-$\beta$-D-fucopyranoside) at range of pH 3 to 10, and high activity using $\beta$-pNPGA ($\rho$-nitrophenyl-$\beta$-D-galactopyranoside) from pH 5 to 10, especially, 3.3 times higher activity at pH 9. Effects of temperature indicated that the $\beta$-glucosidase showed low activity using $\alpha$-pNPG, $\beta$-pNPG, and $\beta$-pNPF from $20^{\circ}C$ to $70^{\circ}C$, and increased activity using $\beta$-pNPGA from $30^{\circ}C$ to $50^{\circ}C$, 1.8 times higher activity at $50^{\circ}C$ than at $30^{\circ}C$. According to activity determination of other substrates, the enzyme was active on daidzin, genistin, and glycitin, inactive on esculin and apigenin-7-glucose. The EDTA and DTT as reducing agents inhibited $\beta$-glucosidase activity, but SDS and mercaptoethanol did not inhibit. Monovalent or divalent metal ions such as $MnSO_4$, $CaCl_2$, KCl, and $MgSO_4$ did not inhibited $\beta$-glucosidase activity. $CuSO_4$ and NaCl showed low inhibition, and $ZnSO_4$ inhibited 3.3 times higher than control.

• The Korean Journal of Pharmacology
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• v.24 no.1
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• pp.111-123
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• 1988

Neutrophil elastases were purified by a three step procedure consiting of one Sephadex G-75 and two HPLC elutions. The elastases cross-reacted with antibodies to human neutrophil elastase. Three bands with molecular weights between 26,000 and 29,700 were observed by gel electrophoresis. At each stage of purification the quantity of Zn increased, reaching molar ratio of 2:1 with elastase in the most purified samples. Calcium content. was seletively elevated during the earlier stages of purification but decreased to a ratio of 0.25 to 1 with elastase at the final step of purfication. Neutrophil elastase could be inhibited by EDTA, EGTA and 1,10-phenanthroline. EGTA inhbition was noncompetitive inhibition and reversible only if the time of preincubation was relatively short, indicating the instability of the apoenzyme. The concentration of chelator required to show significant inhibition of elastase was also dependent upon the stage of purity and the ionic strength of the reaction mixture. Inhibition by EGTA, followed by the removal of EGTA, could be reversed by Zn. In the presence of EGTA the enzyme could be returened to full activity by the addition of Zn, Mn and Ca, but not Mg or Na. All of the above evidence strongly supports human neturophil elastase could be a metalloenzyme as well as a serine protease.

• Korean Journal of Food Science and Technology
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• v.25 no.2
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• pp.185-190
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• 1993

Tilapia cultured in fresh water were acclimated in sea water with daily increase of $5%_{\circ}$ of salinity and it was completely terminated at the 7th day (0 week). Each three tilapia acclimated were examined weekly based from 0 week to elucidate changes of chloride cells in gill, mineral contents and physical properties in muscle and biochemical characteristics in myofibrils. Chloride cells existed in gills were gradually developed in number and size by acclimation to sea water and became to almost constant state at the third week. Shearing value, compressing strength and content of minerals such as Mg, Na and K in muscle were showed remarkable increase by acclimation to sea water in comparison to those of muscle from tilapia reared in fresh water. Myofibrillar $Mg^{2+}-,\;Ca^{2+}-$ and $K^+(EDTA)-ATPase$ activities of tilapia acclimated in sea water also increased showing significant statistic difference (p<0.01) from those of tilapia reared in fresh water However. thermostability of myofibrils was dropped by acclimation to sea water. The increase of shearing value and compressing strength in the muscle of tilapia by acclimation to sea water would be attributed to the increase of myofibrillar ATPase activities which act to accelerate the decomposition rate of ATP. Therefore, it is suggested that this phenomenon associated with muscle contraction could be brought an improvement of texture of tilapia acclimated in sea water.

• Journal of Microbiology and Biotechnology
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• v.30 no.11
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• pp.1659-1669
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• 2020

1,3-α-3,6-anhydro-L-galactosidase (α-neoagarooligosaccharide hydrolase) catalyzes the last step of agar degradation by hydrolyzing neoagarobiose into monomers, D-galactose, and 3,6-anhydro-L-galactose, which is important for the bioindustrial application of algal biomass. Ahg943, from the agarolytic marine bacterium Gayadomonas joobiniege G7, is composed of 423 amino acids (47.96 kDa), including a 22-amino acid signal peptide. It was found to have 67% identity with the α-neoagarooligosaccharide hydrolase ZgAhgA, from Zobellia galactanivorans, but low identity (< 40%) with the other α-neoagarooligosaccharide hydrolases reported. The recombinant Ahg943 (rAhg943, 47.89 kDa), purified from Escherichia coli, was estimated to be a monomer upon gel filtration chromatography, making it quite distinct from other α-neoagarooligosaccharide hydrolases. The rAhg943 hydrolyzed neoagarobiose, neoagarotetraose, and neoagarohexaose into D-galactose, neoagarotriose, and neoagaropentaose, respectively, with a common product, 3,6-anhydro-L-galactose, indicating that it is an exo-acting α-neoagarooligosaccharide hydrolase that releases 3,6-anhydro-L-galactose by hydrolyzing α-1,3 glycosidic bonds from the nonreducing ends of neoagarooligosaccharides. The optimum pH and temperature of Ahg943 activity were 6.0 and 20℃, respectively. In particular, rAhg943 could maintain enzyme activity at 10℃ (71% of the maximum). Complete inhibition of rAhg943 activity by 0.5 mM EDTA was restored and even, remarkably, enhanced by Ca²⁺ ions. rAhg943 activity was at maximum at 0.5 M NaCl and maintained above 73% of the maximum at 3M NaCl. K_m and V_max of rAhg943 toward neoagarobiose were 9.7 mg/ml and 250 μM/min (3 U/mg), respectively. Therefore, Ahg943 is a unique α-neoagarooligosaccharide hydrolase that has cold- and high-salt-adapted features, and possibly exists as a monomer.

• Journal of Microbiology and Biotechnology
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• v.20 no.3
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• pp.518-524
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• 2010

A mannanase gene (man26B) was obtained from a sea bacterium, Paenibacillus sp. BME-14, through the constructed genomic library and inverse PCR. The gene of man26B had an open reading frame of 1,428 bp that encoded a peptide of 475- amino acid residues with a calculated molecular mass of 53 kDa. Man26B possessed two domains, a carbohydrate binding module (CBM) belonging to family 6 and a family 26 catalytic domain (CD) of glycosyl hydrolases, which showed the highest homology to Cel44C of P. polymyxa (60% identity). The optimum pH and temperature for enzymatic activity of Man26B were 4.5 and $60^{\circ}C$, respectively. The activity of Man26B was not affected by $Mg^{2+}$ and $Co^{2+}$, but was inhibited by $Hg^{2+},\;Ca^{2+},\;Cu^{2+},\;Mn^{2+},\;K^+,\;Na^+$, and $\beta$-mercaptoethanol, and slightly enhanced by $Pb^{2+}$ and $Zn^{2+}$. EDTA did not affect the activity of Man26B, which indicates that it does not require divalent ions to function. Man26B showed a high specific activity for LBG and konjac glucomannan, with $K_m,\;V_{max}$, and $k_{cat}$ values of 3.80 mg/ml, 91.70 ${\mu}mol$/min/mg protein, and 77.08/s, respectively, being observed when LBG was the substrate. Furthermore, deletion of the CBM6 domain increased the enzyme stability while enabling it to retain 80% and 60% of its initial activity after treatment at $80^{\circ}C$ and $90^{\circ}C$ for 30 min, respectively. This finding will be useful in industrial applications of Man26B, because of the harsh circumstances associated with such processes.

• Korean Journal of Soil Science and Fertilizer
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• v.48 no.1
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• pp.57-63
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• 2015

Bioavailability of cadmium (Cd) and arsenic (As) can be different depending on soil pH. For this reason, main purpose of this research was to compare bioavailability of Cd and As in agricultural field under varied soil pH and different extractants. Bioavailable fraction of Cd and As in soil was extracted with $CaCl_2$, $NaNO_3$, DTPA, EDTA, and low molecular weight organic acids (LMWOAs). Soil samples and cultivated crops were collected at the range of soil pH 4.5-8.5 and correlation analysis was conducted between bioavailable fraction of Cd and As in soil and total concentration of Cd and As in crops. Results showed that concentration of Cd and As in acidic soil was ranged $0.002-0.462mg\;kg^{-1}$ and $0.041-4.903mg\;kg^{-1}$ respectively. In alkaline condition, concentration of Cd and As were ranged $0.006-0.351mg\;kg^{-1}$ and $0.039-2.807mg\;kg^{-1}$ respectively. Comparing bioavailable fraction of Cd and As in acidic and alkaline soil condition, higher concentration was measured in acidic condition. Similarly, higher average concentration of Cd and Asin crops was observed in acidic condition (0.398 and $0.751mg\;kg^{-1}$ respectively) than alkaline condition (0.248 and $0.264mg\;kg^{-1}$). Among different extractants, LMWOAs method showed higher correlation ($r^2=0.545$) for Cd in acidic condition indicating that LMWOAs method could be applied for evaluating bioavailability of Cd in acidic soil. However, no high correlation was observed for As in both acidic and alkaline condition. Overall, bioavailable fraction of Cd and As can be higher in acidic condition of soil than alkaline condition resulting higher uptake of Cd and As from soil to crops. Therefore, efficient best management practice (BMPs) for Cd and As in acidic soil should be conducted for minimizing uptake of Cd and As into crops.

• Applied Biological Chemistry
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• v.34 no.3
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• pp.225-230
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• 1991

In this study, several combined effect of KCl addition into bring solution, microwave heating of Chinese radish for 3 minutes and addition of two salt mixtures into half fermented Kakdugi were investigated to compare their improving effects of storage stability of Kakdugi, Kakdugi was prepared by salting in 15% NaCl solution for 2 hours and fermented at $25^{\circ}C$ and determined for the changed in pH, total acidity and reducing sugars. It was found from the result that the combined method of salts addition and heating treatment improved the storage stability by 6.4 folds based on pH change where salts mixture of phosphated, Ca EDTA, and Sod. citrate contributed the most effect. Changed in reducing sugar were greatly reduced by salt mixture addition while microwave heating and KCl addition affected little. Storage of Kakdugi at $4^{\circ}C$ after preparation with combined method could extend the storage life more than 8 folds of those Kakdugi stored at $25^{\circ}C$. However total acidity showed higher values for those prepared by combined method and stored at $4^{\circ}C\;or\;25^{\circ}C$.

• Journal of the Korean Society of Food Science and Nutrition
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• v.13 no.3
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• pp.307-312
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• 1984

The dried pure shells deprived of soft tissues were subjected to analysis of chitin-protein complexes from 5 species of marine crustaceans, including 2 species of crabs and 3 species of shrimps. The protein fractions were obtained from chitin-protein complexes under the varying conditions of extractions and the crude chitin was prepared from the shells by the sulfurous acid process. The crude chitin was purified through the extraction with several organic solvents such as dimethyl-acetamide, N-methylpyrrolidone. The purified chitin was also examined using the phase contrast microscope. Total protein contents of the shells were diverse, showing 9.6% for Portunus trituberculatus, 3.1%, Charybdis bimaculata, 9.4%, Penaeus japonicus, 10.9%, Metapenaeus intermedius and 5.8%, Squilla oratoria. Covalently bound protein varied with species from 2.1% for Charybdis bimaculata to 9.9% for Metapenaeus intermedius. The puified chitin contents of the shells were shown to 21.1% for Portunus tritube rculatus, 6.2%, Charybdis bimaculata, 20.2%, Penaeus japonicus, 27.1%, Metapenaeus intermedius and 25.5%, Squilla oratoria. Exceptionally low analytical value obtained with Charybdis bimaculata are supposed to be due to the very young subjects. The ratios of chitin to covalently bound protein in the shells were various such as 2.7 to 1 for Portunus trituberculatus, Penaeus japonicus and Metapenaeus intermedius, 3.1 to 1, Charybdis bimaculata and 6.1 to 1, Squilla oratoria. The microscope finding of the purified chitin showed the filamentous form in all the specimen.

• Journal of the East Asian Society of Dietary Life
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• v.17 no.6
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• pp.808-815
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• 2007

Four different herbs, including rosemary, oregano, basil and sage, along with green tea were individually added to ground beef at different percentages: 0.5, 1.0, and 1.5, respectively. The sensory qualities of the five natural herbs and their antioxidant effects were evaluated using Beef-Yukwonjeon. For the sensory evaluation results, the 0.5% herb addition and the control group generally showed higher acceptability scores than the 1.0% or 1.5% herb addition groups. In particular, the Yukwonjeon made with 0.5% green tea showed higher preferences in the acceptance tests and ranking tests. However, the sensory differences among samples were reduced after 3 days of storage at $6^{\circ}C$. For the results of lipid oxidation without storage, the TBARS values of the herb addition groups were slightly lower than the value of the control group, but not significantly(p>0.05). However, the antioxidant effects of each herb showed higher values over a longer storage period as well as at the higher herb concentrations. The antioxidant effects of the herbs during 7 days' of storage at $6^{\circ}C$ were evaluated as $10.0{\sim}38.5%$ reductions in TBARS values as compared to the group without herbs. Furthermore, the antioxidant effects of the herbs during 30 days' of storage at $-20^{\circ}C$ were shown as $16.8{\sim}27.8%$ reductions in the TBARS value. The green tea showed the highest antioxidant activity against lipid oxidation, because it was acceptable at higher concentrations in the sensory evaluation. These results indicate the potential to produce better quality beef-Yukwonjeon, with herbs, especially green tea.