• Title/Summary/Keyword: trypsin inhibition.

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Characteristics of Trypsin-like Protease and Metalloprotease Associated with Mycelium Differentiation of Streptomyces albidoflavus SMF301

  • Kang, Sung-Gyun;Kim, In-Seop;Jeong, Byung-Cheol;Ryu, Jae-Gon;Rho, Yong-Taik;Lee, Kye-Joon
    • Journal of Microbiology
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    • v.33 no.4
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    • pp.307-314
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    • 1995
  • Trypsin like protease (TLP) and metalloprotease (MTP) were induced in associated with the mycelium differentiation in Streptomyces albidoflavus SMF301. TLP and MTP were purified and characterized from the culture. The molecular mass of TLP and MTP were estimated to be 32 kDa and 18 kDa, respectively. The molecular mass of TLP and MTP were estimated to be 32 kDa and 18 kDa, respectively. The optimum pH and temperature of TLP were 10 and 40.$^{\circ}C$ Those of MTP were 8 and 55 $^{\circ}C$ TLP was stable at alkaline pH (6-9) and unstable above 45.$^{\circ}C$and MTP was stable at alkaline pH and unstable above 80.$^{\circ}C$ Km and Vmax values with benzoyl-arginyl p-nitroanilide of TLP were 139 $\mu$M, and 10 nmole of nitroanilide released per min per$\mu\textrm{g}$ protein, respectively. Km, and Vmax values with a synthetic substrate, leucine p-nitroanilide, or MTP were 58.9 $\mu$M, 3.47 nmol of nitroanilide released per min per$\mu\textrm{g}$protein, respectively. TLP was inhibited competitively by leupeptin; the inhibition constant was 0.0031 $\mu$M. MTP was inhibited by EDTA, phenonthroline and bestatin.

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Plant Phenolics as ${\beta}$-Secretase (BACE1) Inhibitors

  • Jun, Mi-Ra;Lee, Seung-Ho;Choi, Sun-Ha;Bae, Ki-Hwan;Seong, Yeon-Hee;Lee, Kyung-Bok;Song, Kyung-Sik
    • Food Science and Biotechnology
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    • v.15 no.4
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    • pp.617-624
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    • 2006
  • Various plant phenolics were assessed for (${\beta}$-secretase (BACE1) inhibitory activity in order to screen for anti-dementia agents. Among 39 phenolics, eight compounds, 1,2,3-trigalloyl glucopyranoside, acetonyl geraniin, euphorscopin, furosine, helioscopinin A, helioscopinin B, jolkinin, and rugosin E exhibited strong inhibition of BACE1 with $IC_{50}$ values of $5.87{\times}10^{-8}-54.93{\times}10^{-6}\;M$. Among them, rugosin E was the most potent ($IC_{50}$ $5.87{\times}10^{-8}\;M$). The active compounds were shown to be non-competitive inhibitors by Dixon plot. All the phenolic BACE1 inhibitors except furosin also suppressed prolyl endopeptidase (PEP) activity. However, these phenolic compounds caused less inhibition of ${\alpha}$-secretase (tumor necrosis factor a converting enzyme; TACE) and no significant inhibition of other serine proteases such as trypsin, chymotrypsin, and elastase was seen, demonstrating that they are relatively specific to both BACE1 and PEP. No significant structure-activity relationships were found.

Reduction of the Antigenicity of Whey Protein by Enzymatic Hydrolysis (효소가수분해에 의한 유청단백질의 항원성 저하)

  • Ha, Woel-Kyu;Juhn, Suk-Lak;Kim, Jung-Wan;Lee, Soo-Won;Lee, Jae-Young;Shon, Dong-Hwa
    • Korean Journal of Food Science and Technology
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    • v.26 no.1
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    • pp.74-80
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    • 1994
  • As a preliminary study about the reduction of the antigenicity of whey protein isolate(WPI) by treatment of chymotrypsin, trypsin, pancreatin, and protease from Aspergillus oryzae, the properties and antigenicities of whey protein hydrolysates(WPH) were investigated. When degrees of hydrolysis (DH) were measured by use of trinitrobenzensulfonic acid(TNBS), the DH of the WPH treated by pancreatin or protease from Aspergillus oryzae$(5.05{\sim}11.47)$ were much higher than those of the tryptic or chymotryptic WPH$(15.67{\sim}20.20)$. And the pretreatments of heat$(75^{\circ}C)$, 20 min and/or pepsin resulted in higher DH of WPH, generally. When the molecular distributions of the WPH were determined by high performance size exclusion chromatography(HPSEC), the ratios of polypeptides with molecular weight more than 10kDa ranged from 12% to 36%, and the average molecular weights and the average peptide lengths of the WPH were $4,252{\sim}9,132$ dalton and $38{\sim}83$ amino acids, respectively. And there was no bitter taste in all of the WPH. Results of SDS-PAGE showed that most of intact native proteins were eliminated by the enzymatic hydrolysis but there were a few bands of peptides larger than 14.2 kDa in some WPH. When antigenicity was assayed by competitive inhibition enzyme-linked immunosorbent assay(cELISA), monovalent antigenicity of WPH to rabbit anti-WPI antiserum were lowered to $10^{-1.7}-10^{-4.9}$ times and less by the enzymatic hydrolysis. And the pretreatments of heat and pepsin resulted in the lowest antigenicicy within a group of enzymatic hydrolysis, especially in case of the pancreatic hydrolysate(PDP) whose antigenicity was found almost to be removed.

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Inhibitory Substance Produced by Aspergillus sp. on the Snake Venom Proteinase - Isolation of Microorganism and Biological Activities of the Inhibitor - (Aspergillus 속 균주가 생성되는 사독 Proteinase에 대한 저해물질 - 균의 분리 및 저해물질의 생물학적 작용상 -)

  • Hyun, Nam-Joo;Seu, Jung-Hwn
    • Microbiology and Biotechnology Letters
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    • v.15 no.2
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    • pp.129-134
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    • 1987
  • Aspergillus sp. (MK-24) producing a biological active substance that inhibited the venom proteinase activity was isolated from soil. The substance also inhibited the activity of trypsin and coagulation of blood, but did not inhibit papain, $\alpha$-chymotrypsin and pepsin. The substance was partially purified from culture filtrate by precipitaion with acetone, and by chromatography of DEAE-Sepadex A-50 column and Amberlite IRC-50 ion exchange. The inhibitory substance was stable in the wide pH range from 2.0 to 12.0 at 37$^{\circ}C$, but not stable at $65^{\circ}C$ in the alkaline pH. Only 12% of the activity was decreased by the heat treatment at 10$0^{\circ}C$ for two hours. The inhibition on venom proteinase (Agkistrodon bromohoffi brevicaudus) was a mixed type. The inhibitory activity depended on the preincubation time and completely depressed by cupric, zinc and cobalt ions. The inhibition on the venom proteinase was appeared strongly on casein but not on ovalbumin or hemoglobin as a substrate.

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Antimicrobial Characteristics of Metabolites of Lactic Acid Bacteria Isolated from Feces of Newborn Baby and from Dongchimi (신생아 분변 및 동치미에서 분리한 젖산균 대사산물의 항균특성)

  • Lee, Ji-Young;Park, Yeong-Soo;Kim, Yong-Suk;Shin, Dong-Hwa
    • Korean Journal of Food Science and Technology
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    • v.34 no.3
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    • pp.472-479
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    • 2002
  • The antimicrobial effects of metabolites from isolated strains from feces of Korean newborn baby and from Dongchimi against six food-borne microorganisms, and characteristics of the metabolites were investigated.. The metabolites from isolated strains adjusted pH to 3.5, 4.0, and 4.5 showed strong growth inhibition against S. Typhimurium, and S. Enteritidis. The metabolites has kept its inhibition activities to the pathogens after catalase, trypsin or pepsin treatment. In addition, antimicrobial activity of metabolites was not decreased by heat treatment at $121^{\circ}C$ for 15 min. D2 and F35-2 strains were confirmed homofermentative and F20-3 was heterofermentative bacteria identified by final organic acid and gas production. The amount of lactic acid produced by D2 and F35-2 strains after 24 h of incubation was 1.84 and 1.85% respectively, but F20-3 strain produced acetic acid (0.22%) and lactic acid (0.91%).

Amino Acid, Phytic Acid, and Trypsin Inhibitor Contents of Korean Soybeans Before and After Fermentation by Rhizopus oligosporus (Rhizopus oligosporus 발효에 따른 국산 콩의 아미노산, 피틴산 및 트립신 저해제 함량)

  • Hye-Young Park;Hana Song;Hyun-Joo Kim;Min-Jung Seo;Hye Sun Choi;Jiyoung Park;Eun-Yeong Sim;Hong-Sik Kim
    • The Korean Journal of Food And Nutrition
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    • v.36 no.5
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    • pp.415-424
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    • 2023
  • This study investigated the nutritional characteristics of before and after fermentation of domestic soybean (Glycine max L.) by Rhizopus oligosporus. The soybean storage proteins, β-conglycinin (11S globulin) and glycinin (7S globulin), were the most abundant in Seonyu (SY) and Danbaegkong (DBK), with concentrations of 253.4 mg/g and 193.0 mg/g, respectively. For 11S/7S related to sulfur-containing amino acid, DBK had a value of 0.95, making it the most excellent nutritionally among all the cultivars. The free amino acid content significantly increased from 0.04~10.45 mg/g before fermentation to 1.37~16.95 mg/g after fermentation, and the essential amino acid composition increased, confirming an improvement in protein quality after fermentation. Phytic acid, known as a nutritional inhibitor of soybeans, decreased from 1.66~2.13 g/100 g before fermentation to 0.90~1.58 g/100 g after fermentation, suggesting that mineral absorption inhibition was alleviated. In addition, the trypsin inhibitor content is suppressed by 76.20% to 81.25% after fermentation, which is expected to improve protein utilization in the body. This study confirmed some properties of fermented products by Rhizopus oligosporus using domestic soybeans, and these results are presented to serve as the basic data for establishing new uses of Korean soybean cultivars.

Anti-inflammatory Effects, Analgesic Effects, Antipyretic Effects, Anti-oxidative Effects, and Histamine Permeation Inhibition Effects of Modified Gamgil-tang per Extraction Method (추출 방법에 따른 감길탕가미방(甘桔湯加味方)의 항염증(抗炎症), 진통(鎭痛), 해열(解熱), 항산화(抗酸化) 및 Histamine 투과 억제효과)

  • Jin, Hyung-Joon;Kim, Nam-Jae;Choi, Hyuck-Jai;Jung, Hee-Jae;Jung, Sung-Ki
    • The Journal of Internal Korean Medicine
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    • v.29 no.3
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    • pp.554-566
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    • 2008
  • Objectives : Modified Gamgil-tang is a prescription commonly used for respiratory diseases. This thesis was carried out to check the treatment effects and diversity of drug formulation by comparing extraction method of ethanol and water of modified Gamgil-tang. Methods : All experiments were carried out with water and 50% ethanol extraction for comparison. In vivo experiment, hyaluronidase inhibitory effects and trypsin inhibitory effects were tested to measure the anti-inflammatory effects activity. Scavenging effects of DPPH free radical, xanthine oxidase inhibitory effects and inhibition on TBA-RS formation were experimented to measure anti-oxidative effects. With the in vivo experiment, ICR group mice and SD group rats were used as experimental animals. An anti-inflammatory effects experiment were carried out to measure the action on carrageenin-induced hind paw edema: analgesic effects were measured using writhing syndrome induced by 0.7% acetic acid in mice: antipyretic effect was measured using endotoxin, and inhibitory effects of increase vascular permeability induced by 0.5% histamine were measured. Results : For extraction of glycyrrhizin contents, ethanol extract was extracted 2 times of that of water extract. Anti-inflammatory effects showed high in ethanol extract. Anti-oxidative effects measured high in ethanol extract. No significant result was found in inhibition on TBA-RS formation. Analgesic effects were found to be similar in water and ethanol extract. Antipyretic effects were found to be stronger in water extract. Inhibitory effects of increase vascular permeability induced by 0.5% histamine showed stronger in ethanol extract. Conclusion : By measuring anti-inflammatory effects, analgesic effects, antipyretic effects, anti-oxidative effects, and histamine permeation inhibition effects both in water extract and ethanol extract after adding agents such as Mentha Herba, Gardenias Fructus, and propolis to existing Gamgil-Tang, ethanol extract was found to be more effective in anti-inflammatory effects, analgesic effects, anti-oxidative effects, and histamine permeation inhibition effects. The converse was found for antipyretic effect.

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Inhibition of Lipid Accumulation in 3T3-L1 Adipocytes by Different Enzymatic Hydrolysates of Dried Red Sea Cucumber Stichopus japonicus (건조 방법에 따른 홍해삼(Stipchopus japonicus) 효소 가수분해물의 지방 축적 억제 효과)

  • Kim, Seo-Young;Oh, Jae-Young;Kim, Eun-A;Heo, Soo-Jin;Kim, Kil-Nam;Jeon, You-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.53 no.5
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    • pp.707-716
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    • 2020
  • Red sea cucumber Stichopus japonicus, was dried using three methods-far-infrared ray, vacuum, and freeze drying and then enzymatically hydrolyzed using nine proteases: Alcalase, Flavourzyme, Kojizyme, Neutrase, Protamex, trypsin, α-chymotrypsin, and papain. In addition, the potential ability of hydrolysates to inhibit lipid accumulation in 3T3-L1 adipocytes was evaluated. The yield of hydrolysates from red sea cucumbers dried using each method was higher than that of the distilled water extract, and protein contents were either similar or higher. The hydrolysates that exhibited inhibitory effects on lipid accumulation, as demonstrated via Oil red O staining, were those obtained by far-infrared ray drying coupled with Alcalase, Flavourzyme, Kojizyme, or Neutrase treatment. In addition to the advantages of far-infrared drying and the characteristics of Flavourzyme, the Flavourzyme hydrolysate of far-infrared-dried red sea cucumber showed the highest inhibitory effect on lipid accumulation. In addition, this hydrolysate significantly decreased the expression of the protein factor fatty acid-binding protein 4, which is related to the late differentiation of 3T3-L1 adipocytes. Taken together, these results suggest that Flavourzyme hydrolysates from farinfrared-dried red sea cucumber may be used as a functional food and/or a pharmaceutical ingredient for the inhibition of lipid accumulation.

Production of staphylokinase in Staphylococcus hyicus subsp. hyicus strains of swine, poultry and bovine origin (돼지, 닭 및 소유래 Staphylococcus hyicus subsp. hyicus의 staphylokinase 산생능)

  • Park, Joon-seo;Park, Cheong-kyu
    • Korean Journal of Veterinary Research
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    • v.37 no.2
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    • pp.359-365
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    • 1997
  • Staphylococcus hyicus subsp. hyicus strains isolated from pigs, chickens and cattle were examined for the production of staphylokinase after inhibition of staphylococcal proteases by two procedures with EDTA(disodium). In one, EDTA was added to the bovine fibrin-dog plasminogen agar medium in concentration of 0.07% and paper strips soaked in 2mg/ml soy bean trypsin inhibitor were then applied on the agar plates. In the other, paper strips soaked in 5% EDTA solution were applied on the bovine fibrin-dog plasminogen agar plates and the strains to be tested were then streaked at right angles with the strip. By these procedures, staphylokinase activity was detected in 8(88.9%) of 9 strains from diseased pigs and in 57(80.3%) of 71 strains from skin of healthy pigs, but not in any strains from skin of healthy chickens and milk samples of mastitic cattle. Additionally kinase activity in 9 Staphylococcus species and subspecies isolated from bovine intramammary infections was also tested by these procedures. Staphylokinase activity was detected in 74.2% of Staph aureus strains and in 25% of Staph xylosus strains.

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The Physicochemical Properties of $\alpha$-Amylase Inhibitors from Black Bean and Naked Barey in Korea (한국산 검정콩 및 쌀보리 $\alpha$-Amylase 저해물질의 이화학적 특성)

  • 심기환;문주석;배영일
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.3
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    • pp.367-375
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    • 1998
  • The physicochemical properties of the $\alpha$-amylase inhibitors from black bean and naked barley is Korea were investigated. Preincubation time for maximum inhibition was 30min and no activity change was seen after that time. Optimum pH of the $\alpha$-amylase inhibitors from the black bean and naked barley was pH 7.0 and the inhibitory activities were stable in the range of pH 6.0~8.0 in both phosphate and Tris-HCI buffer solutions. Both inhibitors maintained more than 50% of activity after incubation for 17 min at 7$0^{\circ}C$. The inhibitors from the black bean and naked barley maintained more than 50% of activities after treatment for 40 min and 30 min with pepsin, and 30 min and 50 min with trypsin, respectively. Both inhibitors functioned via a noncompetitive mechanism and were active against porcine pancreatic and human salivary $\alpha$-amylases. The activities of both inhibitors were linear for the ionic stength ranging from 0 to 0.9. The addition of 70 mM maltose to the reaction mixture caused a maximum increase in the relative activities of both inhibitors, but it did not affect the dissociation of the EI complex. The activities of both inhibitors were significantly enhanced by adding 1mM of K+ or Mg2+.

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