• Title/Summary/Keyword: Acid protease

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Antioxidative Effects of Food Protein Hydrolysates by Protease (효소(酵素)에 의한 단백질(蛋白質) 가수분해물(加水分解物)의 항산화작용(抗酸化作用))

  • Kim, Seon-Bong;Yeum, Dong-Min;Yeo, Saeng-Gyu;Ji, Cheong-Il;Lee, Yong-Woo;Park, Yeung-Ho
    • Korean Journal of Food Science and Technology
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    • v.21 no.4
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    • pp.492-497
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    • 1989
  • The antioxidant effects against linoleic acid of various protein hydrolysates from fish protein, defatted soybean cake, egg albumin and casein were investigated. Each protein hydrolysate by enzyme hydrolysis exhibited the antioxidative effects by addition of 5mg and 10mg per 1g linoleic arid. Especially, egg albumin and fish protein hydrolysates had a great antioxidative effects. The protein hydrolysates indicated the synergitic effects with ${\alpha}-tocopherol$, and indicated scavenging effects toward metal ion $(Fe^{3+},\;Cu^{2+})$ as prooxidants.

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Isolation and Charaterization of Bioactive Peptides from Hwangtae (yellowish dried Alaska pollack) Protein Hydrolysate

  • Cho, San-Soon;Lee, Hyo-Ku;Yu, Chang-Yeon;Kim, Myong-Jo;Seong, Eun-Soo;Ghimire, Bimal Kumar;Son, Eun-Hwa;Choung, Myoung-Gun;Lim, Jung-Dae
    • Preventive Nutrition and Food Science
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    • v.13 no.3
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    • pp.196-203
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    • 2008
  • Hwangtae, dried Alaska pollack, is a major storage product in the fish processing industry. Hwangtae is prepared by removing the internal organs and drying outdoors during the cold witner months by allowing it to thaw during the daytime and re-freeze at night under sub-zero ($-10^{\circ}C$) conditions and gradually dry from December until the next April for around 5 months from Myungtae. In this study, ground Hwangtae was hydrolyzed using two proteolytic enzymes (pepsin and alcalase) which produced five soluble active peptides from Hwangtae (yellowish dried Pollack, Theragra chalcogramma) protein. Two different peptides with strong antioxidative activity were isolated from the hydrolysate using consecutive chromatographic methods of Sephadex G-25 gel, ion-exchange chromatography on a Sepharose-Sephadex C-25 gel, and high-performance liquid chromatography. The isolated peptides, APO1 and APO2, were composed of 16 and 13 amino acid residues, respectively. Both peptides contained a Gly residue at the C-terminus and the repeating motif Gly-Pro-Hyp. The peptide with a molecular weight less than 1,000 Daltons (APACE) obtained from enzymatic hydrolysates of Hwangtae exhibited the highest ACE inhibitory activity. The APACE peptides was composed of 4 amino acid residues (Gly-Leu-Leu-Pro). These results suggest that Hwangtae hydrolysates could be a good source of peptides with ACE inhibitory activity. Biochemical analysis indicated that two 70 kDa peptides (APG1 and APG2) isolated from the hydrolysate had gelatinoytic activity, which was shown to be a calcium dependent protease type as showed by gelatin SDS PAGE.

Optimization of Enzymatic Treatment for the Production of Hydrolyzed Vegetable Protein (가수분해 식물성 단백질의 효소적 생산을 위한 효소 반응 시스템의 최적화)

  • Chae, Hee-Jeong;In, Man-Jin;Kim, Min-Hong
    • Korean Journal of Food Science and Technology
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    • v.29 no.6
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    • pp.1125-1130
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    • 1997
  • The effects of enzyme combination, pH, acid washing and enzyme treatment sequence were investigated in the hydrolysis of soy protein. Comparing Alcalase vs. Neutrase/Alcalase, it appeared that Neutrase/Alcalase was more efficient than Alcalase alone, as the highest degree of hydrolysis (DH) was seen in Neutrase/Alcalase. A surprisingly high DH (more than 60%) was observed with Flavourzyme in the second hydrolysis. The separation of insolubles from the first hydrolysis had little effect on the second hydrolysis. When the washing water from the first hydrolysis was reused in the next hydrolysis, the DH and protein recovery were increased. The addition of calcium ion showed not so much positive effects by the stabilization of Neutrase on the Protein hydrolysis. The use of carbohydrase and repeated acid washing gave positive effects on DH. The simultaneous treatment using endoprotease and exoprotease with pH adjustment improved DH significantly.

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Effect of Benzalkonium Chloride on Biogas Potential of Pig Slurry (Benzalkonium Chloride가 돈슬러리의 바이오가스 생성에 미치는 영향)

  • Park, H. R.;Choi, H. L.;Suresh, Arumuganainar
    • Journal of the Korea Organic Resources Recycling Association
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    • v.19 no.3
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    • pp.63-72
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    • 2011
  • Benzalkonium chloride is most widely used in S. Korea as a disinfectant in livestock husbandry. Inhibition of biogas potentials were investigated with three different doses of benzalkonium chloride in swine slurry. The system was operated at batch mode. The inhibition rates were 10%, 30~40% and >70% at the dose of 10ppm, 40ppm and 80ppm, respectively assuming it was zero percent in case of no dose. Enzymatic activities were analyzed to determine the enzymatic type which was inhibited by benzalkonium chloride. The acid phosphatase, alkaline phosphatase and protease were shown negatively correlated with biogas potential. Correlation of ${\alpha}$-glucosidase and biogas potentials was observed not high (p<0.01, r=-0.426) while benzalkonium chloride (r=-0.853, p<0.01) and acid phosphatase (p<0.01) with biogas potentials were significantly and negatively correlated. The effect of benzalkonium chloride on Escherichia coli were also evaluated by disc diffusion method. As increase of benzalkonium concentration, inhibition zone of anaerobic bacteria was extended. It revealed that benzalkonium chloride significantly deteriorated biogas potential through inhibition of acetogenic bacteria.

Standardized ileal digestibility of amino acids of protein sources associated with exogenous enzymes for broilers

  • Fortes, Bruno Duarte Alves;Mello, Heloisa Helena de Carvalho;Cafe, Marcos Barcellos;Arnhold, Emmanuel;Stringhini, Jose Henrique
    • Animal Bioscience
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    • v.35 no.7
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    • pp.1030-1038
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    • 2022
  • Objective: Two experiments were conducted to evaluate the effect of enzyme complex (EC) on the standardized ileal digestibility (SID) of amino acids (AA) in corn gluten meal (60%) (CGM), soy protein concentrate (SPC), dried bovine plasma (DBP), and poultry offal meal (POM). Experiments I and II were conducted with broilers in the pre-starter (1 to 7 days of age) and starter (1 to 21 days of age) phases, respectively. Methods: The treatments consisted of a protein-free diet (PFD) containing feedstuffs either supplemented with EC (xylanase, amylase, and protease) or not. In Experiment I, a total of 360 one-day-old male Cobb-500 broiler chicks were randomly housed in 45 pens, resulting in five replicates with eight birds each, totalizing eight treatments and one PFD group. In Experiment II a total of 270 one-day-old male Cobb-500 broiler chicks were randomly housed in 45 pens, resulting in five replicates with six birds each, totalizing eight treatments and one PFD group. The PFD groups were used to assess the endogenous AA losses. The birds were slaughtered to collect the ileal content. Results: In the pre-starter phase, the SID of arginine, branched chain-aminoacids, glycine, serine, aspartate, and glutamic acid increased with EC addition. The EC improved the SID of arginine and glutamic acid of CGM; the SID of valine and cystine of SPC; the SID of leucine, glycine, and aspartate of POM and the SID of isoleucine of DBP. In the starter phase, the SID of isoleucine, phenylalanine and glycine increased in EC-supplemented diets. The EC improved the SID of isoleucine of DBP; the SID of phenylalanine of CGM and POM. The SID of AA of SPC was not influenced by the EC. Conclusion: The addition of an EC to broiler pre-starter and starter diets is efficient in increasing the SID of AA on SPC, POM, and DBP.

Variation of Antimicrobial Peptide in the Extract of the Hard-shelled Mussel Mytilus coruscus Depending on Boiling (가열 유무에 따른 참담치(Mytilus coruscus) 추출물 내의 항균 펩타이드 변화)

  • Lee, Ji-Eun;Seo, Jung-Kil
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.55 no.6
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    • pp.875-885
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    • 2022
  • This study was performed to confirm the optimal extraction method for antimicrobial peptides from the Hard-shelled mussel. Extractions were performed with two processes including 1% HAc/boiling and 1% HAc/non-boiling methods and used extracts for the comparison of the antimicrobial activity, protease stability, action mechanism, AU-PAGE (acid-urea PAGE), and HPLC chromatograms. 1% HAc/boiling extract showed potent antibacterial activities both against Gram-positive and negative bacterium but 1% HAc/non-boiling extract showed antibacterial activity only against Gram-positive bacteria. Treatment of 1% HAc/boiling extract with proteases retained almost antibacterial activity against B. subtilis, but abolished significant antibacterial activity against E. coli D31. Only 1% HAc/boiling extract showed two discrete clearing antibacterial zones including slow migrating and rapid migrating zones. Both extracts showed strong DNA-binding ability but did not show bacterial membrane permeabilizing ability. In comparison of the chromatogram obtained from C18 or cation-exchange HPLC, the eluted peaks from 1% HAc/boiling extract showed high hydrophobic property or absorbance compared to 1% HAc/non-boiling extract, respectively. The concentration of the purified antimicrobial peptide was also higher in 1% HAc/boiling extract than in 1% HAc/non-boiling extract. Our results suggest that the effective extraction condition for antimicrobial peptides from marine invertebrate is boiling process in a weak acetic acid solution (1%).

Antimicrobial Substance against Escherichia coli O157:H7 Produced by Lactobacillus amylovorus ME1

  • Jung, Byung-Moon;Woo, Suk-Gyu;Chung, Kun-Sub
    • Food Science and Biotechnology
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    • v.17 no.3
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    • pp.679-682
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    • 2008
  • A lactic acid bacterium producing an antimicrobial substance against Escherichia coli O157:H7 was isolated from raw milk and identified as Lactobacillus amylovorus ME-1. In addition to E. coli O157 :H7, the antimicrobial substance also inhibited the growth of Bacillus cereus, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus pyrogenes, and Yersinia enterocolitica. The antimicrobial substance was stable at pH 2-12 and $121^{\circ}C$ for 15 min and insensitive to proteinase K, protease, amylase, and catalase. Purification of the antimicrobial substance was conducted through methanol and acetonitrile/ethylacetate extraction, ultrafiltration with a 500 Da cutoff, thin layer chromatography (TLC) with silicagel 60, and high performance liquid chromatography (HPLC) with a $C_{18}$ reverse phase column. The ${\lambda}_{max}$ of the purified antimicrobial substance was determined as 192 nm by ultra violet (UV) scanning, while the molecular weight was estimated as 453 Da based on the mass spectrum. Accordingly, the current results suggest that the antimicrobial substance from the L. amylovorus ME-1 was not a bacteriocin, but rather a new non-proteinaceous substance distinct from acidophilin, acidolin, diacetyl, and reuterin.

Impact of transgenic AFPCHI (Cucumis melo L. Silver Light) fungal resistance melon on soil microbial communities and enzyme activities

  • Bezirganoglu, Ismail;Uysal, Pinar
    • Journal of Plant Biotechnology
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    • v.44 no.2
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    • pp.156-163
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    • 2017
  • A greenhouse experiment was conducted for evaluation of ecological effects of transgenic melon plants in the rhizospheric soil in terms of soil properties, enzyme activities and microbial communities. Organic matter content of soil under transgenic melon plants was significantly higher than that of soil with non-transgenic melon plants. Significant variations were observed in organic matter, total P and K in soil cultivation with transgenic melon plants. There were also significant variations in the total numbers of colony forming units of fungi, actinomycetes and bacteria between soils treated with transgenic and non-transgenic melon plants. Transgenic and non-transgenic melon significantly enhanced several enzymes activities including urease, acid phosphatase, alkalin phosphatase, arysulphtase, ${\beta}$ glucosidase, dehydrogenase, protease and catalase. Soil polyphenoloxidase activity of $T_1$ transgenic melon was lower than that of $T_0$ transgenic melon and a non-melon plant during the same period. The first generation transgenic melon plants ($T_0$) showed significantly greater (p<0.05) effect on the activitiy of arylsulfatase, which increased from $2.540{\times}10^6CFU\;g^{-1}$ (control) to $19.860{\times}10^6CFU\;g^{-1}$ ($T_0$). These results clearly indicated that transgenic melon might change microbial communities, enzyme activities and soil chemical properties.

Development of Methods for Protein Extraction from Three Major Korean Fermented Soy Foods for 2-Dimensional Gel and Mass Spectrometric Analyses

  • Lim, Jin-Kyu
    • Journal of Applied Biological Chemistry
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    • v.51 no.3
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    • pp.88-94
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    • 2008
  • Three different protein extraction methods-phenol extraction, trichloroacetic acid (TCA) precipitation, and desalting/TCA precipitation-were compared to determine the optimal reproducible high resolution 2-dimensional (2-D) electrophoresis for each chungkugjang, doenjang, and kochujang samples. The soluble proteins from Chungkugjang extracted by phenol were separated with high reproducibility and resolution, and gained 1.75- to 3-fold more protein spots on 2-D gel than those from the other methods. On the contrary, the extracted proteins from doenjang and kochujang treated by desalting/TCA precipitation method showed about 1.5- to 3.3-fold more protein spots on 2-D gel. Using the established methods, the changes in the protein profiles of the fermented soy foods were monitored during the fermentation period by 2-DE. One of the major proteins in soy, $\beta$-conglycinin $\alpha$-subuint, and some proteins with unknown functions were localized on 2-D gel as the protease-resistant proteins throughout the fermentation period of doenjang. Changes in the protein profile monitored by the established methods can provide basic information on unfolding the mechanisms of the generation of biofunctional activity in the fermented soy foods.

ORI2 is a Strong Inhibitor of Coxsackievirus B4 Replication (오리방풀로부터 분리된 ORI2의 췌장염 유발 콕사키바이러스B4 증식억제)

  • Lim, Byung-Kwan;Jo, Soyeon;Kim, Jin Hee
    • Korean Journal of Pharmacognosy
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    • v.45 no.4
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    • pp.282-287
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    • 2014
  • The ORI2 (3-[3,4-dihydroxyphenyl]acrylic acid 1-[3,4-dihydroxyphenyl]-2-methoxycarbonylethyl ester) was purified from the extract of Isodon excisus. We confirmed the antiviral effect of ORI2 in a coxsackievirus-induced pancreatitis model. Coxsackievirus B4 (CVB4) is a common cause of pancreatitis and may be reason of the type-1 diabetes. Anti-enteroviral compounds were screened by HeLa cell survival assay. Purified natural compounds were added to HeLa cells cultured 96-well plates after $10^4PFU/ml$ CVB4 pre-incubation for 30 min. ORI2 significantly improved HeLa cell survival in a dose-dependent manner. In addition, ORI2 (1 mM) treatment was dramatically decreased virus protease 2A induced eIF4G-I cleavage and viral VP1 capsid protein production. HeLa cell virus titers and viral RNA replication were significantly decreased in ORI2-treatment in a dose dependent manner (1 mM~0.001 mM). These results demonstrate that ORI2 has a strong antiviral effect. It was significantly decreased virus replication. ORI2 may be developed as a potential therapeutic agent for CVB4.