• Journal of Microbiology and Biotechnology
• /
• v.17 no.8
• /
• pp.1271-1283
• /
• 2007

A fibrinolytic protease (PoFE) was purified from the cultured mycelia of the edible oyster mushroom Pleurotus ostreatus, using a combination of various chromatographies. The purification protocol resulted in an 876-fold purification of the enzyme, with a final yield of 6.5%. The apparent molecular mass of the purified enzyme was estimated to be 32 kDa by SDS-PAGE, fibrin-zymography, and size exclusion using FPLC. The optimal reaction pH value and temperature were pH 6.5 and $35^{\circ}C$, respectively. PoFE effectively hydrolyzed fibrinogen, preferentially digesting the $A{\alpha}$-chain and the $B{\beta}$-chain over the ${\gamma}$-chain. Enzyme activity was enhanced by the addition of $Ca^{2+},\;Zn^{2+},\;and\;Mg^{2+}$ ions. Furthermore, PoFE activity was potently inhibited by EDTA, and it was found to exhibit a higher specificity for the chromogenic substrate S-2586 for chymotrypsin, indicating that the enzyme is a chymotrypsin-like metalloprotease. The first 19 amino acid residues of the N-terminal sequence were ALRKGGAAALNIYSVGFTS, which is extremely similar to the metalloprotease purified from the fruiting body of P. ostreatus. In addition, we cloned the PoFE protein, encoding gene, and its nucleotide sequence was determined. The cDNA of cloned PoFE is 867 nucleotides long and consists of an open reading frame encoding 288 amino acid residues. Its cDNA showed a high degree of homology with PoMEP from P. ostreatus fruiting body. The mycelia of P. ostreatus may thus represent a potential source of new therapeutic agents to treat thrombosis.

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

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

• Journal of Periodontal and Implant Science
• /
• v.30 no.2
• /
• pp.213-231
• /
• 2000

이 연구의 목적은 다공성의 CPP 내부에 쥐의 장골의 골수에서 유래된 세포를 접종하고 3차 원적으로 배양하여 CPP가 골 형성을 위한 조직공학의 지지체로 적용가능한가를 연구하는 것과 Calcium PolyPhosphate(CPP)의 돌연변이 유발성을 검사하는 것이다. 무수 ($Ca(H_2PO_4)$)를 condensation하여 무결정의 ($Ca(PO_3)$)를 얻고 이를 용융하고 냉각시킨 후 분쇄하여 Calcium polyphosphate(CPP) powder를 얻었다. 다공성의 CPP는 5% $SiO_2$를 첨가하여 sponge 형태로 $450-550{\mu}m$ 소공의 크기를 가지는 것과(CPP-45ppi) $200-300{\mu}m$의 소공의 크기를 가지는 것(CCP-60ppi) 2가지로 제작하였다. 각각의 CPP matrices는 $5mm{\times}5mm{\times}1mm$의 블록 형태로 만들었다. 체중 100g 내외의 백서에서 장골(femur, tibia)을 채취하여 백서의 장골 골수 세포를 분리하여 배양한 후 24well에 CPP block을 넣고 CPP block 당 $10^5$개의 배양한 세포를 접종하였다. 배양 1, 7, 14, 및 21 일째에 각 well에서 trypsin EDTA를 이용하여 2회 반복하여 cell을 분리하였고, 원심분리한 후 hemacytometer로 측정하였다. 또, 45ppi와 60ppi, 그 리 고 Tissue Culture Polystyrene(control group)에 접종, 배양된 세포들의 염기성 인산분해효소활성도를 배양 7, 14, 및 21 일째에 각각 측정하였다. 각 기간별로 배양된 세포-CPP 혼합체내에서 세포의 부착 및 증식과 형성된 조직의 3차원적 형태를 관찰하기 위하여 주사전자현미경하에서의 관찰하였다. CPP의 돌연변이 유발성 검사 (mutagenicity test)를 위해 hypoxanthine-guanine phosphoribosyl transferase(HPRT) assay를 하였다. NIH3T3 cell line과 CHO-K1 cell line으로 각각 $1000{\mu}g/m{\ell}$, $100{\mu}g/m{\ell}$, $10{\mu}g/m{\ell}$ 그리고 $1{\mu}g/m{\ell}$의 CPP 농도에서 측정하였다. 통계적 분석을 위해서 모든 측정은 각군당 4개체 이상 시험하였고, 각 측정값은 평균값${\pm}$표준편차로 나타내었다. 각 군간의 통계적 유의성 검정을 위해서 Analysis of variance(ANOVA)를 이용하였고 Tukey의 방법으로 사후분석을 실시하였다. 제작된 CPP matrices 소공들이 서로간에 연결이 잘 되어있는 형태였다. 두 가지로 제조된 CPP(45ppi와 60ppi) 모두에서 세포의 부착이 잘 일어났고, 부착된 세포의 분열도 잘 일어났다. 2 가지의 CPP 모두에서 7, 14, 21일째의 세포 수는 1일째에 비해 유의성 있게 증가하였다(P<0.01). 3차원적 구조인 Calcium PolyPhosphate에서 배양한 세포는 24well dish(tissue culture polystyrene)에서 평면적으로 배양한 대조군의 세포에서 보다 염기성 인산분해효소 (Alkaline Phosphatase)를 유의성 있게 높게 나타냈다. 주사전자현미경에서 세포-CPP 혼합체를 관찰한 결과, CPP block에 세포들이 잘부착되어 있었고, 시간이 지남에 따라 세포가 여러 층을 형성하면서 뭉치는 현상을 보였다. 또, HPRT assay 결과 , Calcium PolyPhosphate는 돌연변이 유발성을 보이지 않았다. 이상의 결과로 볼 때 CPP에는 세포부착이 잘 일어나고, 지지체 상에서 세포의 분열도 활발하게 일어나므로 골조직을 위한 조직공학의 우수한 지지체가 될 수 있을 것으로 사료된다.

• Applied Biological Chemistry
• /
• v.34 no.3
• /
• pp.225-230
• /
• 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 Microbiology and Biotechnology
• /
• v.30 no.11
• /
• pp.1659-1669
• /
• 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.

• Restorative Dentistry and Endodontics
• /
• v.35 no.2
• /
• pp.80-87
• /
• 2010

The purpose of this study was to compare the different canal irrigation methods to prevent the formation of precipitate between sodium hypochlorite (NaOCl) and chlorhexidine (CHX). Extracted 50 human single-rooted teeth were used. The root canals were instrumented using NiTi rotary file (Profile .04/#40) with 2.5% NaOCl and 17% EDTA as irrigants. Teeth were randomly divided into four experimental groups and one control group as follows; Control group: 2.5% NaOCl only, Group 1: 2.5% NaOCl + 2% CHX, Group 2: 2.5% NaOCl + paper points + 2% CHX, Group 3: 2.5% NaOCl + preparation with one large sized-file + 2% CHX, Group 4: 2.5% NaOCl +95% alcohol+ 2% CHX. The teeth were split in bucco-lingual aspect and the specimens were observed using Field Emission Scanning Electron Microscope. The percentages of remaining debris and patent dentinal tubules were determined. Statistical analysis was performed with one-way analysis of variance (ANOVA). Energy Dispersive x-ray Spectroscopy was used for analyzing the occluded materials in dentinal tubule for elementary analysis. There were no significant differences in percentage of remaining debris and patent tubules between all experimental groups at all levels (p > .05). In elementary analysis, the most occluded materials in dentinal tubule were dentin debris. NaOCl/CHX precipitate was detected in one tooth specimen of Group 1. In conclusion, there were no significant precipitate on root canal, but suspected material was detected on Group 1. The irrigation system used in this study could be prevent the precipitate formation.

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

• Korean Journal of Food Science and Technology
• /
• v.39 no.2
• /
• pp.209-216
• /
• 2007

The present study investigated anti-oxidative and anti-inflammatory activity of glycoprotein isolated from Morus Indica Linne (MIL glycoprotein). We found that MIL glycoprotein has a molecular weight of 32 kD and consists of carbohydrate (40.03%) and protein (59.97%), and that it has a strong scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical $({\cdot}OH)$, and superoxide anion $(O_2{\cdot}\;^-)$ radicals. In addition, MIL glycoprotein had a stable character and an optimal DPPH radical scavenging activity in the alkaline and neutral pH solution, and up to at 105. However, the results indicated that it has a minimal scavenging activity in the metal ionic solution ($Ca^{2+}$, $Mn^{2+}$, and $Mg^{2+}$) in the presence of EDTA. In addition, we further investigated whether MIL glycoprotein scavenges oxygen radicals and blocks inflammation-related signals in the bisphenol A (BPA)-stimulated Raw 264.7 cells. The results in this study showed that it has a character to scavenge the productions of reactive oxygen species (ROS) and nitric oxide (NO) dose-dependently. Also it blocked the activities of inflammation-related signals such as nuclear factor-kappa B ($NF-{\kappa}B$) and inducible nitric oxide synthase (iNOS). For example, it had an inhibitory effect on the activation of $NF-{\kappa}B$ (p50) and iNOS proteins at 200 ${\mu}g/mL$ MIL glycoprotein. Here, we speculate that MIL glycoprotein is one of natural antioxidants and of modulators of the BPA-induced inflammation.

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