• BMB Reports
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• v.37 no.6
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• pp.720-725
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• 2004

A number of signaling molecules contain small pleckstrin homology (PH) domains capable of binding phosphoinositides or proteins. Phospholipase C (PLC)-${\gamma}1$ has two putative PH domains, an $NH_2$-terminal (PH1) and a split PH domain ($nPH_2$ and $cPH_2$). We previously reported that the split PH domain of PLC-${\gamma}1$ binds to phosphatidylinositol 4-phosphate (PI(4)P) and phosphatidylinositol 4,5-bisphosphate (PI(4,5)$P_2$) (Chang et al., 2002). To identify the amino acid residues responsible for binding with PI(4)P and PI(4,5)$P_2$, we used site-directed mutagenesis to replace each amino acid in the variable loop-1 (VL-1) region of the PLC-${\gamma}1$ $nPH_2$ domain with alanine (a neutral amino acid). The phosphoinositide-binding affinity of these mutant molecules was analyzed by Dot-blot assay followed by ECL detection. We found that two PLC-${\gamma}1$ nPH2 domain mutants, P500A and H503A, showed reduced affinities for phosphoinositide binding. Furthermore, these mutant PLC-${\gamma}1$ molecules showed reduced PI(4,5)$P_2$ hydrolysis. Using green fluorescent protein (GFP) fusion protein system, we showed that both $PH_1$ and $nPH_2$ domains are responsible for membrane-targeted translocation of PLC-${\gamma}1$ upon serum stimulation. Together, our data reveal that the amino acid residues $Pro^{500}$ and $His^{503}$ are critical for binding of PLC-${\gamma}1$ to one of its substrates, PI(4,5)$P_2$ in the membrane.

• Korean Journal of Microbiology
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• v.46 no.4
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• pp.359-365
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• 2010

Fifty Actinobacteria strains were isolated from rhizosphere soil of Sasa borealis. In the course of screening for antibacterial activity against bacterial leaf spot of pepper (Xanthomonas axonopodis pv. vesicatoria) of isolates, 12 isolates showed strong antibiotic activity. Basis on the 16S rRNA gene sequence, they were belonging to Streptomyces cluster II. Strain JR-24 exhibited strong antibiotic activity against X. axonopodis pv. vesicatoria, had a minimum inhibitory concentration of 10 ${\mu}l$/disc. The strain JR-24 was most closely related to Streptomyces galbus $DSM40089^T$ (98.1%), Streptomyces longwoodensis $LMG20096^T$ (98%) and Streptomyces capoamus $JCM4734^T$ (97.8%). When assayed with the API 20NE and 50 CHE kit, it is positive for utilization of L-arabinose, D-fructose, D-glucose, D-galactose and hydrolysis of gelatin, protein, starch. The strains contained iso-$C_{14:0}$ (25.93%), iso-$C_{15:0}$ (10.13%), anteiso-$C_{15:0}$ (19.29%) and iso-$C_{16:0}$ (20.35%) as major fatty acids and MK-9 (H4), MK-9 (H6), and MK-9 (H8) as the isoprenoid quinone. Strain JR-24 was suggested new species of genus Streptomyces by nearest neighbors of genotypic relationships and phenotypic characterization. This study was important to microbial resources investigation for environment-friendly agriculture.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.31 no.6
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• pp.875-881
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• 1998

To develop functional food material with angiotensin converting enzyme (ACE) inhibitory peptides, muscle protein of anchovy, Engraulis japonica was hydrolyzed during 48 hrs by digestive pretenses such as pepsin, trypsin, $\alpha$-chymotrypsin, and commercial proteases such as papain, bromelain, complex enzyme, Elavourzyme, Novozym, Neutrase, Protamex and Alcalase. The only $50\%$ ethanol soluble hydrolysates were tested for inhibitory activity against ACE and yield of $50\%$ ethanol soluble peptide-nitrogen ($ESPN_{50}$). ACE inhibition effects and yield of $ESPN_{50}$ occurred as hydrolysis time increased to 8 hrs, Among those pretenses tested, hydrolysates by Alcalase and $\alpha$-chymohypsin had greater ACE inhibitory activity (80 and $74\%$, reipectively) with eletated levels of $ESPN_{50}$ (48 and 58 mg/ml, respectively), while Protamex hydrolysates had greater ACE inhibitory activities ($73\%$) with reduced levels of $ESPN_{50}$ (7.2mg/ml) than others. Amino acid compositions of $50\%$ ethanol solubles obtained from those hydrolysates were rich in glutamic acid, aspartic acid, cysteine and leucine.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.5
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• pp.842-849
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• 1997

This study was designed to investigate the antioxidative activity of enzymatic hydrolysates prepared from defatted anchor muscle by various pretenses. In these hydrolysates, the hydrolysates derived from pepsin and Protamex showed the strongest antioxidative activity. Enzymatic hydrolysates also showed the synergistic effects on antioxidative activity of $\alpha-tocopherol$, and almost no change in butylated hydroxytoluene (BHT). Peroxidation of metal ions $(Fe^{3+},\;Cu^{2+})$ was inhibited by enzymatic hydrolysates. Ten fractions (P-1 to P-10) were fractionated from the peptic hydrolysates by Amberlite IR-120 and Bio-gel P-2 column chromatography. The maximum antioxidative activity was observed in the traction P-2 (fraction No. $26\~31$). In amino acid composition of before and after hydrolysis of defatted anchovy muscle and the active fractions (P-2), contents of aspartic arid and glutamic acid were increased, but alanine, cysteine, tyrosine and phenylalanine were decreased.

• 한국해양학회지
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• v.30 no.4
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• pp.341-354
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• 1995

Fluorescence characteristic and amino acids composition of organic matter were determined from extracted seawater samples at eight stations in the East Sea of Korea. Organic compounds have been extracted onto C-18 Sep-Pak cartridges. Three dimensional excitation/emission fluorescence contouring of extracts showed two markedly distinct characterized fluoroscopies representing protein-like biomacromolecule and humic-like geomacromolecule. Protein-like biomacromolecule showing fluorescence maxima at 280 nm/330 nm (excitation/emission) were abundant in the surface mixed layer and then apparently decreased below the thermocline at most stations. It suggests that source of biomacromolecule is comely related with vigorous biological synthetic activity in the surface layer and bacteria decompose its biologically labile components near the thermocline and in the deeper layer. On the other hand, humiliate geomacromolecule showing fluorescence maxima at 330 nm/430 nm (excitation/emission) were low in the surface mixed layer implying photochemical oxidation and then increased below the thermocline at most stations. It suggests that geomacromolecule might be transformed by condensation of bio-refractoryorganic fraction after decomposition of biomacromolecule and particulate organic carbon derived from the surface mixed layer. HPLC measurements of amino acids showed similar composition between seawater and extracted organic macromolecule after hydrolysis. Glycine, serine and alanine were predominant, accounting for more than 50% of total amino acids. Dissolved free amino acids of seawater were more abundant in the surface layer(0.7∼1.8 uM) than the deeper layer (0.2∼0.4 uM). D/L racemic ratio of alanine of extracted organic matter showed lower value in the surface layer than the deeper layer. It suggests that biomacromolecule predominant in the surface layer is relatively young, rapidly recycling and biologically labile.

• Journal of Chest Surgery
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• v.27 no.3
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• pp.177-186
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• 1994

Pentastarch is a hydroxyethyl starch similar to hetastarch, but lower average molecular weight and fewer hydroxyethyl groups which result in enhanced enzymatic hydrolysis and faster renal elimination.This report was performed to compare the clinical efficacy and safety of 10 % pentastarch[Pentaspan , group I] for plasma volume expansion after open heart surgery with that of 5% albumin[Plasmanate, group II]. There were no statistically significant differences between the group I [n=18] and group II [n:19] in the preoperative parameters [age, sex, body weight] and operative parameters[bypass time, aorta cross clamping time]. During the first 24 hours after arrival of the patient in the surgical intensive care unit, colloid solution [500--1000 ml] was infused to maintain left atrial pressure of more than 8 mmHg, or cardiac index of 2.0 L/min/M2 of more. In results, there were 3 complications of hypotension immediately after infusion of 5 % albumin solution and 2 among the 3 patients were excluded for the study. However there was no complication after infusion of 10 % pentastarch solution. Hemodynamic responses to infusion was similar for both groups, although in group I a greater increase in both left atrial pressure[mean 1.8 versus 0.7 mmHg, p< 0.05] and right atrial pressure [mean 2.2 versus 1.7 mmHg, p < 0.05] was observed during infusion of the first 500 ml. There were no significant differences in any of the measured respiratory parameters[PaO2, intrapulmonary shunt, and effective lung compliance]. Homodilution with colloid significantly reduced hemoglobin [mean 1.2 versus 0.8 gm/dl], and serum protein and albumin level[total protein;4.8$\pm$ 0.5 versus 5.2 $\pm$0.5 gm/dl, p < 0.05: albumin: 3.2 $\pm$0.4 versus 3.6 $\pm$0.6 gm/dl, p < 0.05] by 6:00 AM on 1 day postoperatively, however there were no significant differences on 7 day postoperatively. The mean serum colloid osmotic pressure and osmolarity was similar in both group.There were no abnormal findings of liver function and kidney function in all the patients. There were no significant between-group differences in bleeding time, platelets, prothrombin time, activated partial thromboplastin time and amount of chest tube output measured on 1st and 7th postoperative day. These findings demonstrated that 10% pentastarch is more effective and safe for plasma volume expension than 5 % albumin solution with no adverse effects on coagulation. Also 10 % pentastarch is less expensive than 5 % albumin and it would appeare to be a reasonable first choice for plasma volume expansion.

• Food Science and Biotechnology
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• v.18 no.1
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• pp.95-102
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• 2009

The aerial parts of garland (Chrysanthemum coronarium L.) were extracted in 80% aqueous methanol (MeOH) and the concentrated extract was then partitioned using ethyl acetate (EtOAc), n-butanol (n-BuOH), and $H_2O$, successively. EtOAc and n-BuOH fractions resulted in 4 glycerides with the application of octadecyl silica gel and silica gel column chromatography. The chemical structures of the glycerides were determined using several spectroscopic methods, including nuclear magnetic resonance (NMR) and mass spectrometry (MS) as (2S)-1-O-palmitoyl-sn-glycerol (1), (2S)-1-O-oleoyl-2-O-oleoyl- 3-O-$\beta$-D-galactopyranosyl-sn-glycerol (2), (2S)-1-O-palmitoyl-2-O-linoleoyl-3-O-phosphorouscholine-sn-glycerol (3), and (2S)-1-O-linolenoyl-2-O-palmitoyl-3-O-[$\alpha$-D-galactopyrasyl-($1{\rightarrow}6$)-$\beta$-D-galactopyranosyl]-sn-glycerol (4). The free fatty acids of these glycerides were determined with gas chromatography (GC)-MS analysis following alkaline hydrolysis and methylation. These glycerides demonstrated an inhibitory effect on acyl-CoA: cholesterol acyltransferase (ACAT, compound 1: $45.6{\pm}0.2%$ at $100{\mu}g/mL$), diacylglycerol acyltransferase (DGAT, compound 1: $59.1{\pm}0.1%$ at $25{\mu}g/mL$), farnesyl protein transferase (FPTase, compound 2: $98.0{\pm}0.1%$; compound 3: $55.2{\pm}0.1%$ at $100{\mu}g/mL$), and $\beta$-secretase ($IC_{50}$, compound 4: $2.6{\mu}g/mL$) activity. This paper is the first report on the isolation of these glycerides from garland and their inhibitory activity on ACAT, DGAT, FPTase, and $\beta$-secretase.

• Journal of the Korean Society of Food Science and Nutrition
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• v.25 no.5
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• pp.779-785
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• 1996

The qualities of bread and change of phytic acid during breadmaking with whole wheat flour were investigated. The ratios of ash contents in wheat flour and whole wheat flour were 0.41% and 1.57%, respectively. The ratios of fiber contents in wheat flour and whole wheat flour were 0.14% and 1.83%, respectively. In amino acid analysis, glutamic acid was determined to be 32~36g/100g protein, which was the highest. Lysine, glycine, arginine and aspartic acid were higher in whole wheat flour than those of wheat flour. Proline, glutamic acid, and phenylalanine were higher in wheat flour than those of whole wheat flour. The ratio of phytic acid content in wheat flour and whole wheat flour was 0.312% and 0.734%, respectively. The content of phytic acid during beadmaking was decreased approximately 65% after proofing, while this was almost constant in the process of oven baking. The content of phytic acid in bread with 3% yeast had less hydrolysis than that in bread with 5% yeast during breadmaking. The phytic acid content in the 0.1% yeast food was decreased more than the 0, 0.3, and 0.5% yeast food groups. As the amount of whole wheat flour increased, the volume of bread was decreased, and color became dark. The sensory evaluation was showed the quality of bread to be the highest when the amounts of coarse whole wheat flour and fine whole wheat flour was 20% and 30%, respectively. Though the amount of coarse whole wheat flour and fine whole wheat flour were increased up to 30% and 50%, respectively, external characteristics of bread was remained in normal.

• Food Science and Preservation
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• v.24 no.1
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• pp.107-115
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• 2017

The aim of this study was to investigate the hepatoprotecive effect of silk protein hydrolysates (SDH), which was prepared by acid hydrolysis, in rats. SDH itself did not exhibit any cytotoxic effect on hepatic tissues. SDH showed a protective effect on tert-butyl hydroperoxide (t-BHP)-induced hepatotoxicity and liver damage. SDH effectively reduced AST (aspartate aminotransferase) and ALT (alanine aminotransferase), which are biomarkers for liver damage, in a dose-dependent manner. Malondialdehyde (MDA), a lipid peroxidation product, was significantly reduced by SDH. A high dose of SDH (2 g/kg) reduced t-BHP-induced MDA production by 40%. Glutathione (GSH), which is an endogenous antioxidant molecule, was effectively increased by SDH treatment. GSH content was enhanced by around 2.5-fold, compared with t-BHP control, upon SDH (2 g/kg) treatment. Lactate dehydrogenase (LDH), which is an enzyme released by cell cytotoxicity, was greatly increased by t-BHP, but significantly decreased by SDH treatment. Furthermore, hematoxylin and eosin (H&E) staining showed that SDH suppressed t-BHP-induced lesions in liver tissue. Taken together, SDH might be used as a protective agent against liver damage.

• Journal of Life Science
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• v.16 no.7 s.80
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• pp.1235-1242
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• 2006

Sulforaphane, an isothiocyanate derived from hydrolysis of glucoraphanin in broccoli and other cruciferous vegetables, was shown to induce phase II detoxification enzymes and inhibit chemically induced mammary tumors in rodents. Recently, sulforaphane is known to induce cell cycle arrest and apoptosis in human canter cells, however its molecular mechanisms are poorly understood. In tile present study, we demonstrated that sulforaphane acted to inhibit proliferation and induce morphological changes of human hepatocarcinoma HepG2 cells. Treatment of HepG2 cells with $10{\mu}M\;or\;15{\mu}M$ sulforaphane resulted in significant G2/M cell cycle arrest as determined by DNA flow cytometry. Moreover, $20{\mu}M$ sulforaphane significantly induced the population of sub-G1 cells suggesting that sulforaphane induced apoptosis. This anti-proliferative effect of sulforaphane was accompanied by a marked inhibition of ryclin A, cyclin 31 and Cdc2 protein. However, the levels of tumor suppressor p53 and Cdk inhibitor p21 mRNA and protein expression were significantly increased by sulforaphane treatment in a concentration-dependent manner. Although further studies are needed, the present work suggests that sulforaphane may be a potential rhemoprevetiveichemotherapeucc agent for the treatment of human cancer cells.