• Journal of Microbiology and Biotechnology
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• v.20 no.4
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• pp.700-707
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• 2010

A novel antioxidative peptide (APVPH I, antioxidative peptides from venison protein hydrolysates I) was purified from venison by enzymatic hydrolysis, column chromatography of DEAE-Sephacel, and high-performance liquid chromatography. The molecular mass of the purified peptide was found to be 9,853 Da and the amino acid sequences of the purified peptide was Met-Gln-Ile-Phe-Val-Lys-Thr-Leu-Thr-Gly. The purpose of this study was to evaluate the effects of APVPH I against $H_2O_2$-induced neuronal cells damage in PC-12 cells. Antioxidative enzyme levels in cultured neuronal cells were increased in the presence of the peptide. In addition, APVPH I inhibited productions of nitric oxide (NO), reactive oxygen species (ROS), malondialdehyde (MDA), and cell death against $H_2O_2$-induced neuronal cell damage in PC-12 cells. It was presumed to be APVPH I involved in regulating the apoptosis-related gene expression in the cell environment. The present results indicate that APVPH I substantially contributes to antioxidative properties in neuronal cells.

• Toxicological Research
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• v.17
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• pp.97-104
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• 2001

Three types of proteases (MEF-1, MEF-2 and MEF-3) were purified from the egg cases of Ten-odera sinensis using ammonium sulfate fractionation, gel filtration on Bio-Gel P-60 and affinity chromatography on DEAE Affi-Gel blue gel. The proteases were assessed homogeneous by SDS-polyacrylamide gel electrophoresis and have molecular weight of 31,500, 32,900 and 35,600 Da, respectively. The N-terminal regions of the primary structure were compared and they were found to be different each other. MEFs readily digested the $A\alpha$ - and B$\beta$-chains of fibrinogen and more slowly the ${\gamma}$-chain. The action of the enzymes resulted in extensive hydrolysis of fibrinogen and fibrin, releasing a variety of fibrinopeptides. MEF-1 was inactivated by Cu$^{2+}$ and Zn$^{2+}$ and inhibited by PMSF and chymostatin. MEF-2 was inhibited by PMSF, TLCK. soybean trypsin inhibitor. MEF-3 was only inhibited by PMSF and chymostatin. Antiplasmin was not sensitive to MEF-1 but antithrombin III inhibited the enzymatic activity qf MEF-1. MEF-2 specifically bound to anti plasmin Among the chromogenic protease substrates, the most sensitive one to the hydrolysis of MEFs was benzoyl-Phe-Val-Arg-p-nitroanilide with maximal activity at pH 7.0 and 3$0^{\circ}C$. MEF-1 preferentially cleaved the oxidized B-chain of insulin between Leu15 and Tyr16. In contrast, MEF-2 specifically cleaved the peptide bond between Arg23 and Gly24. D-dimer concentrations increased on incubation of cross-linked fibrin with MEF-1, indicating the enzyme has a strong fibrinolytic activity.ity.

• Korean Journal of Medicinal Crop Science
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• v.13 no.4
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• pp.150-153
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• 2005

Although each part of Trichosanthes kirilowii is frequently used as medicinal herbs, study on the chemical composition is not sufficient. It was found that sarcocarp consists of 70% of carbohydrate, 13% of crude protein, 5% of crude fat, 6% of crude fiber and 6% of crude ash; seed consists of 62.59% of carbohydrate, 12.75% of crude protein, 14.80% of crude fat, 6.50% of crude fiber and 3.36% of crude ash; and root consists of 89.40% of carbohydrate, 4.10% of crude protein, 0.50% of crude fat, 3.50% of crude fiber and 2.50% of crude ash. Sarcocarp and seed contain fifteen kinds of amino acids such as Asp, Ser, Glu, Gly, His, Arg, Thr, Ala, Pro, Try, Cys, Met, Val, Leu, and Phe. Sarcocarp contain large quantities of phenylalanine and glycine and seed does not contain leucine but lysine. The mineral contents in Trichoxanthes kirilowii are 0.55% Ca, 0.91% Mg, 10.29% Na, and 0.17% K.

• Molecular & Cellular Toxicology
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• v.5 no.2
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• pp.160-164
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• 2009

Notch signaling plays a crucial role in development of the nervous system. Neurodevelopmental hypothesis on etiology of schizophrenia has been implicated. The aim of this study is to determine whether single nucleotide polymorphisms (SNPs) of Notch homolog 4 (Drosophila) (NOTCH4) gene are associated with schizophrenia. This study included 283 schizophrenia patients diagnosed according to DSM-IV and 301 normal control subjects. Control subjects without history of psychiatric disorders were recruited. Four missense SNPs [rs915894 (exon 3, Lys117Gln), rs2071282 (exon 4, Pro204Leu), rs422951 (exon 6, Thr320Ala), and rs17604492 (exon 18, Gly942Arg)] of NOTCH4 gene were genotyped by the direct sequencing method. Multiple logistic regression models (codominant, dominant, and recessive models) were employed to evaluate odds ratio, 95% confidence interval, and p value. For analysis of genetic data, SNPStats, Haploview, HapAnalyzer, SNPAnalyzer, and Helixtree programs were also used. Of 4 SNPs, rs2071282 was weekly associated with schizophrenia in two alternative models (codominant model, P=0.049; dominant, P=0.041). However, these associations were not significant after Bonferroni correction. At 4 SNPs, one linkage disequilibrium (LD) block was made. This block consisted of rs915894 and rs2071282. In haplotype analysis, AC haplotype was weakly associated with schizophrenia (dominant, P=0.04). This association was disappeared after Bonferroni correction. Our result shows possibility that some SNPs of NOTCH4 gene may be weekly associated with development of schizophrenia in Korean population. However, replication result by other population will be needed.

• Food Science of Animal Resources
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• v.37 no.3
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• pp.402-409
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• 2017

A novel peptide having free radical scavenging activity was separated, using an on-line high-performance liquid chromatography (HPLC) - ABTS screening method, from bovine skim milk fermented by Lactococcus lactis SL6 (KCTC 11865BP). It was further purified using reverse phase-HPLC (RP-HPLC) and sequenced by RP-HPLC-tandem mass spectrometry. The amino acid sequence of the identified peptide was determined to be Phe-Ser-Asp-Ile-Pro-Asn-Pro-Ile-Gly-Ser-Glu-Asn-Ser-Glu-Lys-Thr-Thr-Met-Pro-Leu-Trp (2,362 Da), which is corresponding to the C-terminal fragment of bovine ${\alpha}_{s1}$-casein (f179-199). The hydroxyl radicals scavenging activity ($IC_{50}$ $28.25{\pm}0.96{\mu}M$) of the peptide chemically synthesized based on the MS/MS data showed a slightly lower than that of the natural antioxidant Trolox ($IC_{50}$ $15.37{\pm}0.52{\mu}M$). Furthermore, derivatives of the antioxidant peptide were synthesized. The antioxidative activity of the derivatives whose all three proline residues replaced by alanine significantly decreased, whereas replacement of two proline residues in N-terminal region did not affect its antioxidative activity, indicating that $3^{rd}$ proline in C-terminal region is critical for the antioxidative activity of the peptide identified in this study. In addition, N-terminal region of the antioxidant peptide did not show its activity, whereas C-terminal region maintained antioxidative activity, suggesting that C-terminal region of the peptide is important for antioxidative activity.

• Clinical and Experimental Pediatrics
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• v.59 no.sup1
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• pp.45-48
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• 2016

Short-chain acyl-CoA dehydrogenase deficiency (SCADD) is a rare autosomal recessive mitochondrial disorder of fatty acid ${\beta}$-oxidation, and is associated with mutations in the acyl-CoA dehydrogenase (ACADS) gene. Recent advances in spectrometric screening for inborn errors of metabolism have helped detect several metabolic disorders, including SCADD, without symptoms in the neonate period. This allows immediate initiation of treatment and monitoring, so they remain largely symptomless metabolic disease. Here, we report a 15-month-old asymptomatic male, who was diagnosed with SCADD by newborn screening. Spectrometric screening for inborn errors of metabolism 72 hours after birth revealed an elevated butyrylcarnitine (C4) concentration of $2.25{\mu}mol/L$ (normal, < $0.99{\mu}mol/L$). Urinary excretion of ethylmalonic acid was also elevated, as detected by urine organic acid analysis. To confirm the diagnosis of SCADD, direct sequencing analysis of 10 coding exons and the exon-intron boundaries of the ACADS gene were performed. Subsequent sequence analysis revealed compound heterozygous missense mutations c.164C>T (p.Pro55Leu) and c.1031A>G (p.Glu344Gly) on exons 2 and 9, respectively. The patient is now growing up, unretarded by symptoms such as seizure and developmental delay.

• Journal of Nutrition and Health
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• v.8 no.1
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• pp.47-59
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• 1975

Free amino acids in extracts and total amino acids in hydrolysates of eleven species of edible mushrooms were analyzed by amino acid autoanalyzer (Technicon PNC-1 Type). All these 11 species of mushroom can be repesented for convenience sake as follows. S-1; Agaricus campestris Fr. S-2: Agaricus campestris S-3; Pholiota nameko(I. Ito) S. Ito et Imai S-4; Auricularia auricula-judae(Fr.) $Qu{\acute{e}}l$ S-5; Tremella fuciformis Berk. S-6; Tricholoma matsutake(S. Ito et Imai) Sing. S-7; Pleurotus ostreatus Fr. $Qu{\acute{e}}l$ S-8; Lentinus edodes Berk Sing. S-9; Ramaria botrytis (Pers.) Ricken S-10; Coprinus comatus(Fr.) S.F, Gray S-11; Gyrophora esculenta The results obtained from this study are as follows. 1) 17 kinds of amino acid, including 7 kinds of essential amino acid in human nutrition except tryptophan were identified and quantified. 2) Of all free amino acids contained in mushrooms, glutamic acid is the richest, and then comes Ala, Thr, Pro and Lys in that order. There were no found Cys'and His in S-9;His in S-1; Met and Arg in S-11; Cys and Met in S-5;Pro, Cys, Met, Lys and Arg in S-4. Of all total amino acids which are closely related with nutritional valuation, glutamic acid is the richest, and then comes Asp, Ala, Arg, Leu, Thr, Gly in that order. Especially S-1 and S-2 contain high quantity $o{\acute{i}}$ proline in both free and total amino acids. 3) Cotents of ammonia in extracts of mushrooms in decreasing order in S-1, S-10, S-8, S-2, S-7, S-6, and S-2, S-6, S-8, S-9, S-1 in hydrolysates of mushrooms. 4) Gross Contents of free amino acid in extracts is high in decreasing order in S-10, S-1, S-7, S-6, S-8, and total amino acid in hydrolysates is high in S-10, S-2, S-2, S-8, S-1, S-9, S-6. 5) Besides 17 kinds of amino acid, 5 kinds of unknown amino acid are found in extracts and hydrolysates.

• Journal of the Korean Society of Food Science and Nutrition
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• v.15 no.4
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• pp.1-9
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• 1986

The mashes of soybean paste were preparea using the conventional meju fermented naturally by wild microoganisms or the new types of meju fermented by pure cultures of Aspergillus oryzae, Bacillus natto and B. subtilis to elucidate changes during the aging period. The results obtained are as follows ; The soybean paste made with conventional meiu and Asp. oryzae meju showed higher content of amino nitrogen than those of B, natto and B. subtilis meju. Soybean paste made with conventional meju contained a little more content of total and reducing sugars than other soybean pastes. ph during aging period was higher than 5.0 for the Asp. oryzae paste while less than 4.5 for B. subtilis paste. Aspartic acid. threonine, serine, glutamic acid, glycine, alanine, cystine, valine, methionne, leucine and histidine for Asp. oryzae paste ; tyrosine, arginine and proline for conventional meju paste; and isoleucine and phenylalanine for B. subtilis paste were found to be peak amount 90 days after the preparation. The content of total free amino acid was high in the order of Asp. oryzae paste, conventional paste, B. natto paste and B. subtilis paste.

• Molecules and Cells
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• v.19 no.1
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• pp.97-103
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• 2005

The role of residue C323 in catalysis by human glutamate dehydrogenase isozymes (hGDH1 and hGDH2) was examined by substituting Arg, Gly, Leu, Met, or Tyr at C323 by cassette mutagenesis using synthetic human GDH isozyme genes. As a result, the $K_m$ of the enzyme for NADH and ${\alpha}-ketoglutarate$ increased up to 1.6-fold and 1.1-fold, respectively. It seems likely that C323 is not responsible for substrate-binding or coenzyme-binding. The efficiency ($k_{cat}/K_m$) of the mutant enzymes was only 11-14% of that of the wild-type isozymes, mainly due to a decrease in $k_{cat}$ values. There was a linear relationship between incorporation of [$^{14}C$]p-chloromercuribenzoic acid and loss of enzyme activity that extrapolated to a stoichiometry of one mol of [$^{14}C$] incorporated per mol of monomer for wild type hGDHs. No incorporation of [$^{14}C$]p-chloromercuribenzoic acid was observed with the C323 mutants. ADP and GTP had no effect on the binding of p-chloromercuribenzoic acid, suggesting that C323 is not directly involved in allosteric regulation. There were no differences between the two hGDH isozymes in sensitivities to mutagenesis at C323. Our results suggest that C323 plays an important role in catalysis by human GDH isozymes.

• Journal of Pharmaceutical Investigation
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• v.26 no.1
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• pp.43-53
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• 1996

The objective of this study was to determine whether 4-phenylazobezyloxycarbonyl-Pro-Leu-Gly-Pro-D-Arg (Pz-peptide), an enhancer of hydrophilic solute permeability in the intestine, could elevate the paracellular permeability of hydrophilic drugs across cornea and conjunctiva in the rabbit. The in-vitro penetration of hydrophilic drugs (mannitol, atenolol) and lipophilic drug (propranolol) across the rabbit cornea and conjunctiva was studied either in the presence or absence of 3 mM Pz-peptide. Drug penetration was evaluated using the modified Ussing chamber. The conjunctiva was more permeable than the cornea to all drugs. Pz-peptide showed enhanced effects on the drug transport across cornea and conjunctiva in a concentration dependent manner. Effects or ion transport inhibitor on the mannitol penetration were then investigated. Mannitol penetration was not changed by serosal addition of $100\;{\mu}M$ ouabain, suggesting that $Na^+/K^+$ ion tranporter was not involved in the Pz-peptide induced elevation of paracellular drug permeability. Furthermore, effects of Pz-peptide and EDTA on the transport of atenolol and propranolol into the ocular tissues or blood circulation after its administration into both eyes were investigated. EDTA showed enhanced effect on propranolol transport into the ocular tissues, but Pz-peptide did not show significant difference. Systemic absorption of propranolol by the addition of EDTA or Pz-peptide was not changed. On the other hand, EDTA and Pz-peptide elavated the atenolol transport into the ocular tissues. The transport of atenolol into the blood circulation was also enhanced by the addition of EDTA, but no effect was observed by the addition of Pz-peptide. The above findings suggest that Pz-peptide would be used as an paracellular pathway enahncer of hydrophilic drugs into the eye, without affecting the systemic absortion of topically applied opthalmic drugs.