• Title/Summary/Keyword: trypsin activity

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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.

Hyaluronidase Inhibitory and Antioxidant Activities of Enzymatic Hydrolysate from Jeju Island Red Sea Cucumber (Stichopus japonicus) for Novel Anti-aging Cosmeceuticals

  • Ding, Yuling;Jiratchayamaethasakul, Chanipa;Kim, Eun-A;Kim, Junseong;Heo, Soo-Jin;Lee, Seung-Hong
    • Journal of Marine Bioscience and Biotechnology
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    • v.10 no.2
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    • pp.62-72
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    • 2018
  • An active ingredient with hyaluronidase (HAse) inhibitory effect is one of the anti-aging approaches in cosmeceuticals. Here, red sea cucumbers (RSCs), Stichopus japonicus, from Jeju Island were evaluated to examine their HAse inhibitory and antioxidant activity effect. In this study, RSCs were extracted by six enzymatic hydrolysis (Alcalase; Al, Trypsin; Try, Neutrase; Neu, Pepsin; Pep, Alpha-chymotrypsin; Chy and Protamex; Pro). Alcalase hydrolysate (AlH) showed the highest antioxidant capacities for both of oxygen radical absorbance capacity (ORAC) and trolox equivalent antioxidant capacity (TEAC) methods, compared to those of other hydrolysates, at $66.59{\pm}0.78{\mu}M\;TE/mg$ and $135.78{\pm}3.24{\mu}M\;TE/mg$, respectively. Furthermore, AlH performed the highest capacity of HAse inhibitory with $IC_{50}$ value of 3.21 mg/ml. Thus, RSCs hydrolyzed by Al were chosen to determine the cellular antioxidant activity and hyaluronic acid (HA) production effect on Human immortalized keratinocyte cell line (HaCaT). The results showed that AlH improved the cell viabilities and intracellular reactive oxygen species (ROS) induced by 2,2'-Azobis(2-amidinopropane) dihydrochloride (AAPH) were significantly decreased. In addition, AlH increased HA amount by regulating HYAL2 and HAS2 expressions in the HaCaT cells. Taken together, AlH of RSCs collected from Jeju Island showed HAse inhibitory and antioxidant activities against skin-aging which shows its potentials can be an optional natural bioactive ingredient for novel cosmeceuticals.

Effect of Sodium Caseinate Hydrolysates on Angiotensin-I Converting Enzyme Inhibition Activity (Sodium Caseinate 가수분해물의 Angiotensin-I Converting Enzyme 저해효과에 관한 연구)

  • Lee, Keon-Bong;Shin, Yong-Kook;Baick, Seung-Chun
    • Food Science of Animal Resources
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    • v.32 no.5
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    • pp.652-658
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    • 2012
  • This study was carried out to identify the ACE (Angiotensin converting enzyme) inhibitory activity of casein hydrolysates for development of anti-hypertensive hydrolysates. Sodium caseinate was treated with six kinds of commercial proteases such as Flavourzyme, Protamex, Neutrase 1.5, Alcalase, Protease M, and Protease S for 8 h individually, and was then treated with the enzyme combination for 4 h at $45^{\circ}C$. The hydrolysate which had the highest ACE inhibitory effect was then hydrolysed successively with three digestive enzymes: pepsin, trypsin, and ${\alpha}$-chymotrypsin, at $37^{\circ}C$ for 4 h under conditions mimicking those of the gastrointestinal tract. UF (ultra filtration) treatment was applied to one of the secondary hydrolysates to determine ACE inhibitory activity. When sodium caseinate was hydrolysed by commercial proteases, the degree of hydrolysis (DH) showed 2.54 to 4.25% and after secondary hydrolysis, DH showed 4.30 to 5.22%. ACE inhibitory activity and $IC_{50}$ values decreased, and inhibition rates increased during hydrolysis. Protamex treatment showed the lowest $IC_{50}$ value ($516{\mu}g/mL$) and Flavourzyme hydrolysate showed the highest $IC_{50}$value ($866{\mu}g/mL$). As the first hydrolysate was treated with Flavourzyme, the ACE inhibitory activity increased. Neutrase hydrolysate had the highest activity with an $IC_{50}$ value ($282{\mu}g/mL$). When Neutrase plus Flavourzyme treatment was hydrolyzed by digestive enzymes, the $IC_{50}$ value ($597{\mu}g/mL$) was decreased statistically (p<0.05). As Neutrase plus Flavourzyme hydrolysate is treated by UF with MW cut-off 10,000, permeate showed $273{\mu}g/mL$ of $IC_{50}$ value, showed no difference, but retentate which has over MW 10,000 showed statistically different $IC_{50}$ value, $635{\mu}g/mL$ (p<0.05).

Quality Changes of Dried Lavers during Processing and Storage 3. Changes in Pigments, Trypsin Indigestible Substrates(TIS) and Dietary Fiber Content during Roasting and Storage (김의 가공 및 저장중의 품질변화 3. 배소 및 저장중의 색소, Trypsin 저해물질(TIS) 및 Dietary Fiber의 변화)

  • LEE Kang-Ho;RYUK Ji-Hee;JEONG In-Hak;JUNG Woo-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.23 no.4
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    • pp.280-288
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    • 1990
  • Quality stability of dried lavers during roasting and storage was investigated by measuring the changes of pigment contents including chlorophyll a, carotenoids and biliproteins, the content of trypsin indigestible substrates(TIS), in vitro apparent protein digestibility, and dietary fiber. In heat treatment or roasting of dried laver, carotenoids and chlorophyll a were found to be more stable than biliproteins. Chlorophyll a and carotenoids were retained more than $85\%$ during roasting for 1 hour at $120^{\circ}C$ while biliproteins were retained only $10\%$ at the same temperature. The in vitro digestility of dried layers tended to increase with raising the roasting temperature. The in vitro digestibility of $85\%$ for the roasted laver at $100^{\circ}C$ was higher than that observed in the control of $80\%$. There was a correlation between the decrease in TIS and biliproteins as the laver was roasted. The soulble dietary fiber(SDF) content was substantially increased by heat treatment. The extent of protein digestiblility appeared to be related to the increase of SDF content. In the storage of roasted lavers under both water activities 0.1 and 0.65, the loss of the pigments and TlS were markedly retarded at Aw 0.1. Chlorophyll a was retained about $20\%$ at aw 0.65 and $75\%$ at aw 0.1 after 20 week sto-rage. At worst, more than $90\%$ of the carotenoids were lost at aw 0.65 after 20 week, while biliproteins were comparatively stable at the same water activity. TIS decreased about $15\%$ and in vitro apparent protein digestibility increased up to $92\%$ at aw 0.65 during storage.

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Effects of fermented soybean meal with Bacillus velezensis, Lactobacillus spp. or their combination on broiler performance, gut antioxidant activity and microflora

  • Tsai, C.F.;Lin, L.J.;Wang, C.H.;Tsai, C.S.;Chang, S.C.;Lee, T.T.
    • Animal Bioscience
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    • v.35 no.12
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    • pp.1892-1903
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    • 2022
  • Objective: A series of experiment were conducted to evaluate the effects of replacing a part of soybean meal (SBM) at 6% of broiler diets with fermented soybean meal (FSBM) obtained by single or two-stage fermentation by measuring growth performance, antioxidant activity in the jejunum and distal intestinal microflora. Methods: Soybean meal samples were prepared by single-stage fermentation using Bacillus velezensis (Bv) (FSBMB), or Lactobacillus spp. (as commercial control) (FSBML). Additional SBM sample was prepared by two-stage fermentation using Bv and subsequently using Lactobacillus brevis ATCC 367 (Lb) (FSBMB+L). Enzyme activity, chemical composition, trichloroethanoic acid-nitrogen solubility index (TCA-NSI) and antioxidant activity were measured. Then, in an in vivo study, 320 Ross308 broilers were divided into four groups with ad libitum supply of feed and water. Four groups were fed either a corn-soybean meal diet (SBM), or one of fermented SBM diets (FSBMB+L, FSBMB, and FSBML). Growth, serum characteristics, microflora, and the mRNA expression of selected genes were measured. Results: Compared to SBM, FSBMB+L contained lower galacto-oligosaccharide, allergic protein, and trypsin inhibitor, and higher TCA-NSI by about three times (p<0.05). Reducing power and 1,1-diphenyl-2-picrylhydrazyl free radical scavenging ability correlated positively with the TCA-NSI content in FSBM. Growth performances were not significantly different among four groups. In jejunum of 35-day-old broilers, partial replacement of SBM by FSBMB+L increased the activity of superoxide dismutase and catalase (CAT), and the FSBMB group had the highest catalase activity (p<0.05). Partial replacement of SBM by FSBM increased relative mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and peptide transporter 1 (PepT1) (p<0.05); however, FSBMB+L increased CAT mRNA level to 5 times of the control (p<0.05). Conclusion: Using Bv- and Lb-processed SBM through two-stage fermentation to partially replace 6% of diets will improve the gut's antioxidant activity under commercial breeding in broilers.

Antimicrobial Activity of Lactobacillus reuteri Against Major Food-Borne Pathogens (Lactobacillus reuteri의 주요 식품 위해 미생물에 대한 항균 효과)

  • Kwon, Nam-Hoon;Kim, So-Hyun;Bae, W.K.;Kim, J.Y.;Lim, J.Y.;Noh, K.M.;Kim, J.M.;Ahn, J.S.;Hur, J.;Park, Y.H.
    • Journal of Food Hygiene and Safety
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    • v.16 no.4
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    • pp.264-273
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    • 2001
  • Antimicrobial activities of ave different probiotics (Lactobacillus reuteri, L. acidophillus, L. bulgaricus, L. casei and Bifidobacterium longum) against 8 bacterial pathogens were determined on the Mueller Hinton Agar containing supernatant of probiotics obtained from 3 different growth conditions (MRS without glycerol, MRS with 0.5 M glycerol or 0.25 M glycerol solution). Though antimicrobial activity of L. reuteri in the first two conditions was not better than the others', the activity was significantly higher than that of others in 0.25 M glycerol solution. This prominent effect might be attributable to reuterin, produced by L. reuteri using glycerol. We could detect the presence of reuterin in the supernatant of 0.25 M glycerol solution with 500 MHz Nuclear Magnetic Resonance (NMR). The result of minimum bactericidal concentration (dilution fold) has revealed that reuterin showed pan-bactericidal effects against 8 major food-borne pathogens. To examine any changes of antimicrobial activities of the probiotics, the probiotics were treated with different pH conditions, pepsin or trypsin digestion. Antimicrobial activity of reuterin was not entirely affected by any of these treatments, while the activities of the other probiotics were significantly decreased.

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Calcium-binding Peptides Derived from Tryptic Hydrolysates of Cheese Whey Protein

  • Kim, S.B.;Lim, J.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.10
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    • pp.1459-1464
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    • 2004
  • The purpose of this research was to investigate the potential use of cheese whey protein (CWP), a cheese by-product. The physiological activity of calcium-binding peptides in CWP may be used as a food additive that prevents bone disorders. This research also examined the characteristics of calcium-binding peptides. After the CWP was heat treated, it was hydrolyzed by trypsin. Then calcium-binding peptides were separated and purified by ion-exchange chromatography and reverse phase HPLC, respectively. To examine the characteristics of the purified calcium-binding peptides, amino acid composition and amino acid sequence were analyzed. Calcium-binding peptides with a small molecular weight of about 1.4 to 3.4 kDa were identified in the fraction that was flowed out from 0.25 M NaCl step gradient by ion-exchange chromatography of tryptic hydrolysates. The results of the amino acid analysis revealed that glutamic acid in a calcium-binding site took up most part of the amino acids including a quantity of proline, leucine and lysine. The amino acid sequence of calcium-binding peptides showed Phe-Leu-Asp-Asp-Asp-Leu-Thr-Asp and Ile-Leu-Asp-Lys from $\alpha$-LA and Ile-Pro-Ala-Val-Phe-Lys and Val-Tyr-Val-Glu-Glu-Leu-Lys from ${\beta}$-LG.

PIGEON PEA AS A RUMINANT FEED

  • Cheva-Isarakul, B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.5 no.3
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    • pp.549-558
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    • 1992
  • The study on the potential use of pigeon pea (PP) as a ruminant feed was carried out with sheep in 3 experiments. Digestibility of dry pigeon pea leaves (PPL) and pigeon pea seeds (PPS) determined by differential and regression methods respectively, with rice straw (RS) as a basal diet, revealed that PPS contained higher nutritive value and palatability than PPL. On dry matter (DM) basis, PPL and PPS contained 19.8 and 20.0% CP, 7.3 and 2.3% EE, 6.0 and 4.4% ash, 61.1 and 51.7% NDF, and 29.4 and 17.5% ADF, respectively. The trypsin inhibitor activity in the seed was 3 times of that in the leaves (19.5 vs 7.0 mg TIA/g DM). The digestibility of PPL and PPS were 50.2 and 72.2% in DM, 52.7 and 73.3% in OM, 51.0 and 65.1% in CP respectively. DM intake as well as the digestibility of most nutrients increased with the increasing level of PPS. Digestible energy (DE), Total digestible nutrient (TDN) and N-balance of sheep fed solely PPS, estimated by regression method, was 3.2 kcal/g, 71.1% and 6.3 g/d respectively. Pigeon pea seeds can be well used to substitute soybean meal in concentrate rations for ruminants or directly supplemented to low quality roughages.

Characterization of an antioxidant peptide from katsuobushi (dried bonito) protein hydrolysates

  • Lee, Jung Kwon;Jeon, Joong-Kyun;Byun, Hee-Guk
    • Journal of Marine Bioscience and Biotechnology
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    • v.7 no.1
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    • pp.19-27
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    • 2015
  • The objective of the current study was to evaluate the inhibitory and antioxidant activities of powdered katsuobushi (dried bonito) protein hydrolysates and their corresponding fractions. The powdered katsuobushi (dried bonito) hydrolysates were obtained by enzymatic hydrolysis using Alcalase, ${\alpha}$-chymotrypsin, Neutrase, pepsin, papain, and trypsin. The antioxidant efficacy of the respective hydrolysates were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl, superoxide, and alkyl radical-scavenging activities. Among the hydrolysates, the peptic-derived hydrolysate exhibited the highest antioxidant activity compared to other enzymatic hydrolysates. Therefore, the peptic-derived hydrolysate was further analyzed, and was found to contain an active peptide with an amino acid sequence identified as Pro-Met-Pro-Leu-Asn-Ser-Cys (756 Da). The purified peptides from powdered katsuobushi (dried bonito) had an $EC_{50}$ value of $105.82{\mu}M$, and exhibited an inhibitory effect against DNA oxidation induced by hydroxyl radicals. Taken together, these results suggests that powdered katsuobushi (dried bonito) could be used as a natural antioxidant in functional foods and prevent oxidation reactions in food processing.

Tissue- and Reproductive Organ-specific Expression of Protease Nexin-1 in Sprague-Dawley Rat (흰쥐에서 단백질 분해효소 저해제, Nexin-1의 조직 및 생식기관 특이적 유전자 발현)

  • 고정재;김남근;김진규;최명진;정형민;서승염;김윤희;이현환;차광열
    • Development and Reproduction
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    • v.2 no.2
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    • pp.135-140
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    • 1998
  • Protease Nexin-1 (PN-1) inhibits the activity of several serine proteases including thrombin, urokinase (uPA)-type plasminogen activator and trypsin. Tissue- and reproductive organ-specific mRNA levels of the PN-1 were investigated in Sprague-Dawley adult rat. PN-1 mRNA expression in rats was found in brain (forebrain, hindbrain), heart, liver, lung, ovary and oviduct. The level of PN-1 mRNA in male and female among the tissues was the highest in forebrain of the female. PN-1 expression in reproductive organs was found only in ovary and oviduct. These results suggest that PN-1 expression is dependent on the sex and may be related to folliculogenesis and early embryogenesis.

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