• Title/Summary/Keyword: trypsin inhibitor.

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Characterization of a New Antidementia $\beta$-Secretase Inhibitory Peptide from Rubus coreanus

  • Lee, Dae-Hyoung;Lee, Dae-Hyung;Lee, Jong-Soo
    • Food Science and Biotechnology
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    • v.17 no.3
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    • pp.489-494
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    • 2008
  • In order to develop a potent antidementia $\beta$-secretase inhibitor from phytochemicals, $\beta$-secretase inhibitory activities of extracts from many medicinal plants and herbs were determined. Water extracts from Rubus coreanus showed the highest $\beta$-secretase inhibitory activity of 84.5%. After purification of the $\beta$-secretase inhibitor from R. coreanus using systematic solvent extraction, ultrafiltration, Sephadex G-10 column chromatography, and reverse-phase high performance liquid chromatography (HPLC), a purified $\beta$-secretase inhibitor with $IC_{50}$ inhibitory activity of $6.3{\times}10^3\;ng/mL$ ($1.56{\times}10^{-6}\;M)$ was obtained with a 0.08% solid yield. The molecular mass of the purified $\beta$-secretase inhibitor was estimated to be 576 Da by liquid chromatography-mass spectrometry (LC-MS) and $\beta$-secretase inhibitor also is a new tetrapeptide with the sequence Gly-Trp-Trp-Glu. The purified $\beta$-secretase inhibitory peptide inhibited $\beta$-secretase non-competitively and also show less inhibition on trypsin, however no inhibition on other proteases such as $\alpha$-secretase, chymotrypsin, and elastase.

Use of Chemical Treatments to Reduce Tannins and Trypsin Inhibitor Contents in Salseed (Shorea robusta) Meal

  • Mahmood, S.;Khan, Ajmal M.;Sarwar, M.;Nisa, M.;Lee, W.S.;Kim, S.B.;Hur, T.Y.;Lee, H.J.;Kim, H.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.9
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    • pp.1462-1467
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    • 2007
  • This study investigated the effect of chemical treatments on tannins (condensed and hydrolysable) and on the trypsin inhibitor (TI) activity in salseed meal. Triplicate samples of ground salseed meal (1 kg) were mixed with 820 ml of either distilled water (pH 5.3), 0.67 M acetic acid (pH 2.4), 0.67 M sodium bicarbonate (pH 8.2) or 2% polyvinyl-pyrrolidone (PVP) solution. The material was placed in airtight plastic containers and incubated at $37^{\circ}C$ for 0, 3, 6, 12, 24, 48 and 72 h. Samples of untreated salseed meal which had not been subjected to soaking or incubation were run through the analysis to serve as control. Addition of water, acetic acid, sodium bicarbonate and PVP solutions to salseed meal and subsequent anaerobic incubation at $37^{\circ}C$ significantly reduced chemically detectable tannins. At each incubation time, alkali solution was more effective than its counterparts. The effect of acidic solution on hydrolysable tannin was least among the treatments. All the treatments reduced TI activity of salseed meal. The reduction in TI activity by these treatments was similar and ranged between 80-84%. Treatment time effected a decrease in the contents of antinutritional substances. However, the effect of the treatment with the reagents, even for zero incubation time, was quite pronounced. It may be concluded from the present results that the treatment of salseed meal with sodium bicarbonate (0.67 M) is more effective in reducing hydrolysable and condensed tannin contents than PVP, water and acid solutions. Treatment with sodium bicarbonate solution is more economical and easier to handle than acid and PVP treatments. Incubation of the treated material for 12 h is reasonably effective, economical and safe from any mould growth.

Antimicrobial and Antioxidative Activities of Solvent Fraction from Humulus japonicus (환삼덩굴의 용매분획별 항균성 및 항산화성)

  • Park, Seung-Woo;Woo, Cheol-Joo;Chung, Shin-Kyo;Chung, Ki-Taek
    • Korean Journal of Food Science and Technology
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    • v.26 no.4
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    • pp.464-470
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    • 1994
  • The biological activities of Humulus japonicus were extracted by water and methanol. Methanol was better solvent than water in the extraction for antimicrobial activities against six different species of bacteria and two yeasts. The methanol extract was systematically fractionated with various organic solvents which have different polarities. From the result of antimicrobial activities against six species of bacteria and two species of yeasts, methanol extract was superior to water extract. The methanol extract of Humulus japonicus showed antimicrobial activity against the all species of microorganisms tested except Escherichia coli . The butanol fraction of methanol extract showed antimicrobial effect on the all species tested. The minimal inhibition concentration(MIC) of the butanol fraction on the growth of microorganisms was ranged between $0.1{\sim}0.4%$. The water extract of Humulus japonicus did not show inhibition of the activity of trypsin but methanol extract showed inhibitory activity. The chloroform fraction of methanol extract showed comparatively higher trypsin inhibitory activity than other fractions. The concentration of 50% inhibition$(IC_{50})$ by chloroform fraction was 1.0 mg/ml. Enzyme-inhibitor complex formation was above 90% of the while for 20 min. It was revealed that methanol extract of Humulus japonicus inhibited peroxide production of lard and soybean oil as substrate by antioxidative test. The chloroform fraction of methanol extract had the highest activity. When 0.2% of chloroform fraction was added, induction period of soybean oil and lard were extended 15, 9 days, respectively.

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Purification and characterization of a trypsin inhibitor from egg of skipjack tuna, Katsuwonus pelamis

  • Kim, Se-Kwon;Park, Joung-Ho;Je, Jae-Young
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2001.05a
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    • pp.148-149
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    • 2001
  • Protein inhibitors are proteins or peptides capable of inhibiting catalytic activities of proteolytic enzymes. They are grouped primarily as either serine, cysteine, aspartic or metallto-proteinase inhibitors. Pretense inhibitors have been hewn since the end of the last century in nematodes and human blood serum, and their ubiquitous distribution in microorganisms, animals and plants has been widely documented. (omitted)

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Identification of Proteins in Human Follicular Fluid by Proteomic Profiling

  • Sim, Young-Jin;Lee, Mi-Young
    • Molecular & Cellular Toxicology
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    • v.4 no.3
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    • pp.253-259
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    • 2008
  • Human follicular fluid (HFF) is the in vivo microenvironment for oocyte maturation and includes a variety of proteins that could be involved in oocyte development and fertilization. We therefore used a proteomic approach to identify new HFF proteins. HFF from mature human follicles was obtained from five women following oocyte collection for in vitro fertilization (IVF). Ethanol-precipitated HFF run on two-dimensional gel electrophoresis (2DE) produced approximately 250 Coomassie brilliant blue-stained spots, 64 of which were identified using matrix-assisted laser desorption/ionization-mass spectrometry (MALDIMS). In this study, several proteins including complement factor H, inter-${\alpha}$ (globulin) inhibitor H4, inter-${\alpha}$-trypsin inhibitor heavy chain H4 precursor, human zinc-${\alpha}$-2-glycoprotein chain B, PRO2619, PRO02044, and complex-forming glycoprotein HC were new proteins that have not been previously reported in HFF using proteomic methods. Additionally, we identified alloalbumin venezia for the first time from trichloroacetic acid (TCA)-precipitated HFF. These HFF proteins could serve as new biomarkers for important human reproductive processes.

Purification and Properties of Protease Inhibitor from Streptomyces sp. SK-862 (방선균이 생성하는 단백질 가수분해효소 저해물질의 정제 및 특성)

  • 김중배
    • The Korean Journal of Food And Nutrition
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    • v.11 no.6
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    • pp.678-682
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    • 1998
  • A strain of Streptomyces sp. SK-862, isolated from soil in Wonju city, was able to prodce a biologically active substance that has a strong inhibitory activity against proteolsis by trypsin. The inhyibitory substance was extracted by n-butanol, and then purified by the adsorption chromatography followed by the reverse-phase high performacne liquid chromatography. The purified substance was stable over the pH range from 2 to 10, but was unstable when treated at 8$0^{\circ}C$ for 60 min. This substance was soluble in water, methanol, ethanol nd butanol, but insoluble in chlorofrom and ethylacetate. The Rf value of the purified substance on the thin layer chromatography were 0.56 in n-butanol : methanol : water(5 : 3 : 1v/v) solvent system compare dto 0.23 in ethanol : ammonium hydoxide : water(8 : 1 : 1v/v) solvent system. This substance has maximum absorption at 259 nm. The chemical reaction of the substance was negative for sugar but positive for ninhydrine and iodine reaction.

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The Contents of Antinutritional Factors and Lipoxygenase Activity of the Recommended Soybean Varieties in Korea (장려품종 콩의 영양저해 인자 및 리폭시게나아제 특성)

  • Kim, Dong-Man;Baek, Hyung-Hee;Kim, Kil-Hwan
    • Korean Journal of Food Science and Technology
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    • v.22 no.4
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    • pp.393-397
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    • 1990
  • The contents of the several antinutritional factors and lipoxygenase activity of 19 soybean varieties recommended for cultivation in Korea were analyzed. The ranges of raffinose and stachyose contents, which are the flatulence factors, were $0.74{\sim}1.58%\;d.b.\;and\;3.34{\sim}5.30%d.b.$, respectively and the total amount of these sugars was high in the varieties of Hill, Baekun and Jangbaek. The contents of trypsin inhibitor and phytate phosphorus in the soybean varieties ranged from 21.2 to 37.0 TI/g, d.b. and from 337 to 605mg%, d.b. respectively. The lipoxygenase activity of $163.6{\sim}403.5unit$ was shown in the 19 soybean varieties, and Padal, Jangbaek and S-133 were the varieties with the higest activity.

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Accumulation of triple recessive alleles for three antinutritional proteins in soybean with black seed coat and green cotyledon

  • Kang, Gyung Young;Choi, Sang Woo;Chae, Won Gi;Chung, Jong Il
    • Journal of Plant Biotechnology
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    • v.47 no.2
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    • pp.118-123
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    • 2020
  • The black seed coat of soybeans contain anthocyanins which promote health. However, mature soybean seeds contain anti-nutritional factors like lipoxygenase, lectin and Kunitz Trypsin Inhibitor (KTI) proteins. Furthermore, these seeds can be used only after the genetic elimination of these proteins. Therefore, the objective of this study was to develop novel soybean genotypes with black seed coat and triple recessive alleles (lx1lx1lx2lx2lx3lx3, titilele) for lipoxygenase, lectin, and KTI proteins. From a cross of parent1 (lx1lx2lx3/lx1lx2lx3, ti/ti, Le/Le) and parent2 (lx1lx2lx3/lx1lx2lx3, Ti/Ti, le/le), 132 F2 seeds were obtained. A 3:1 segregation ratio was observed during F2 seed generation for the inheritance of lectin and KTI proteins. Between a cross of the Le and Ti genes, the observed independent inheritance ratio in the F2 seed generation was 9: 3 : 3 : 1 (69 Le_Ti_: 32 leleTi_: 22 Le_titi: 9 leletiti) (χ2=2.87, P=0.5 - 0.1). From nine F2 seeds with triple recessive alleles (lx1lx1lx2lx2lx3lx3, titilele genotype), one novel strain posessing black seed coat, and free of lipoxygenase, lectin and KTI proteins, was selected. The seed coat color of the new strain was black and the cotyledon color of the mature seed was green. The weight of 100 seeds belonging to the new strain was 35.4 g. This black soybean strain with lx1lx1lx2lx2lx3lx3, titilele genotype is a novel strain free of lipoxygenase, lectin, and KTI proteins.

Screening System for Chitin Synthase II Inhibitors from Natural Resources and its Inhibitor Prodigiosin

  • Hwang, Eui-Il;Kim, Young-Kook;Lee, Hyang-Bok;Kim, Hong-Gi;Kim, Sung-Uk
    • Journal of Microbiology and Biotechnology
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    • v.10 no.2
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    • pp.251-257
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    • 2000
  • Chitin synthases are identified as key enzymes of chitin biosynthesis in most of the fungi. Among them, chitin synthase II has been reported to be and essential enzyme in chitin biosynthesis, and exists as a membrane-bound form. To search and screen new antifungal agents from natural resources to inhibit chitin synthase II, the assay conditions were established using the enzyme isolated from Saccharomyces cerevisiae ECY38-38A(pAS6) that overproduces only chitin synthase II. This enzyme was activated only by partial proteolysis with trypsin. Its actibity reached the maximum at $80{\;}\mu\textrm{g}/ml$ of trypsin and was strongly stimulated by 2.0 mM $Co^{2+}$, 1.0 nM UDP-[$^{14}C$]-GicNAc, and 32 mM free-GlcNAc. Under these assay conditions, the highest chitin synthase II activity was observed by incubation at $30^{\circ}C$ for 90 min. However, and extremely narrow range of organic solvents up to as much as 25% of DMSO and 25% of MeOH was useful for determining optimal assay conditions. After a search or potent inhibitors of chitin synthase II from natural resources, prodigiosin was isolated from Serratia marcescens and purified by solvent extration and silica gel column chromatographies. The structure of prodigiosin was determined by UV, IR, Mass spectral, and NMR spectral analyses. Its molecular weight and formula were found to be 323 and $C_{20}H_{25}N_{3}O$, respectively. Prodigiosin ingibited chitin synthase II by 50% at the concentration of $115{\;}\mu\textrm{g}/ml$.

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