• Title/Summary/Keyword: trypsin.

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Drying and Stabilization of Deer Blood (생녹혈의 건조 및 안정화)

  • Ahn, Yong-Geun
    • The Korean Journal of Food And Nutrition
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    • v.22 no.1
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    • pp.20-28
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    • 2009
  • According to traditional oriental medicine, only non-coagulated native deer blood is said to be effective, and coagulated deer blood is ineffective. Thus, a drying and tablet-producing method for deer blood was developed to maintain its physiological and therapeutic activity, and so that after drying, it can be redissolved and protected from coagulation. Proteases such as trypsin, pepsin, chymotrypsin, and aminopeptidase were added to the deer blood indicating that it coagulated in an hour, as shown by the reference. Wax gourd extract, which is high in protease, was added to the blood resulting in anticoagulation for 31 hours. Also, additions of 1% EDTA, 0.38% sodium citrate, 0.16% calcium oxalate, 1.2% ethanol, and 0.006% heparin to the deer blood resulted in anticoagulation for 1 hour, 4 hours, 2 hours, 1 hour, and 31 hours, respectively. In an experiment using 0.19% sodium citrate plus 1% wax gourd extract, and 0.006% heparin plus 1% wax gourd extract, anticoagulation was maintained for up to 72 hours. However, since heparin can not be used in food, the deer blood tablet was made with the addition of 0.19% sodium citrate and 1% wax gourd extract, followed by freeze drying. The dissolution rate for the tablet manufactured in this manner was 96.7%. And the dissolution rates for spray-dried deer blood, vacuum-dried deer blood, and marketed deer blood tablets were 85%, 81%, and 25.5%, respectively. The composition of the tablet produced from the freeze-dried deer blood was 56.4% protein, 18.7% lactose, 1.2% amino acids, 1.0% glucose, 0.7% lipids, 180 mg/100 g of iron, 13 mg/100 g of potassium, 39.1 mg/100 g of calcium, 480 mg/100 g of sodium, 368 mg/100 g of chloride, each.

Fermentation of rice flour with Weissella koreensis HO20 and Weissella kimchii HO22 isolated from kimchi and its use in the making of jeolpyeon (김치유산균(Weissella koreensis HO20, Weissella kimchii HO22)으로 발효한 쌀가루의 이화학적 특성 및 이를 이용한 절편의 제조)

  • Choi, Hyejung;Lee, Hwawon;Yoon, Sun
    • Korean journal of food and cookery science
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    • v.29 no.3
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    • pp.267-274
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    • 2013
  • Demand for a rice cake, a popular traditional food in Korea, is rising, but its industrial-scale production is extremely difficult due to its short shelf-life caused by starch retrogradation and microbial spoilage. By means of the sourdough fermentation technique, we attempt to develop rice cakes with a longer shelf-life. Heterofermentative lactic acid bacteria (Weissella koreensis HO20, Weissella kimchii HO22) isolated from kimchi were used to ferment wet-milled rice flour for their abilities to produce exopolysaccharides and to inhibit the microbial spoilage of rice cakes. After 24 hr of fermentation at $25^{\circ}C$, viable cell counts in rice dough increased from $10^6$ CFU/g to $10^8$ CFU/g and total titratable acidity increased from 0.05% to 0.20%, whereas pH decreased from 6.5 to 5.1. Fermented rice flour showed significantly lower peak, trough, and final viscosities as well as breakdown and setback viscosities measured by rapid viscoanalyzer. Both lactic acid bacteria showed in vitro antifungal activity against Penicillium crustosum isolated from rice cakes. The antifungal activity remained constant after the treatments with heat, proteinase K and trypsin, but fell significantly by increase of pH. Rice cakes made of fermented rice flour were found to retard mycelial growth of P. crustosum. The degree of retrogradation as measured by the hardness of the rice cake was significantly reduced by the use of fermented rice flour. The results suggest that use of fermented rice flour has a beneficial role in retarding starch retrogradation and in preventing fungal growth, hence extending the shelf-life of rice cakes.

The Development of Gastrointestinal Tract and Pancreatic Enzymes in White Roman Geese

  • Shih, B.L.;Yu, B.;Hsu, J.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.6
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    • pp.841-847
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    • 2005
  • The objective of this experiment was to investigate the development of gastrointestinal tract and activities of pancreatic enzymes in White Roman geese. Thirty developing embryos at the 22th, 24th and 26th day of incubation and at hatching, and sixteen or eight goslings, half males and half females, at the 1, 3, 7 or 11, 14, 21 and 28 days of age were sampled, respectively. The weights of the yolk, gastrointestinal tract and intestinal length, and the activities of pancreatic enzymes were measured. Residual yolk weight decreased rapidly during late incubation and was nearly depleted at 3 days of age. The protein and energy contents in the residual yolk of goslings at 3 days of age were significantly (p<0.05) less than those at the late incubation. From 6 days before hatching to 28 days of age, the absolute weights of gizzard, proventriculus, liver, pancreas, small intestine and large intestine in goslings increased by 48, 457, 94, 2334, 89 and 76 times, respectively. The relative weights of proventriculus, gizzard, liver, pancreas, small intestine and large intestine reached peaks at 3, 3, 14, 14, 11 and 11 days of age, respectively, and then decreased gradually. However, the relative lengths of small intestine and large intestine reached peaks at 3 days of age and at hatching, respectively. The activities of pancreatic trypsin and chymotrypsin increased sharply from hatching to 14 day of age, and then decreased gradually until 21 days of age. The activity and specific activity of pancreatic amylase were increased following by age and peaked at 7 to 11 and 21 days of age, respectively. The activity and specific activity of pancreatic lipase reached a plateau from 11 to 28 days of age. These results indicate that the gastrointestinal tract and activities of pancreatic enzymes developed more rapidly than body weight through the early growing period of goslings.

Roasting and Extruding Affect Nutrient Utilization from Soybeans in 5 and 10 kg Nursery Pigs

  • Kim, I.H.;Hancock, J.D.;Hines, R.H.;Gugle, T.L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.2
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    • pp.200-206
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    • 2000
  • Ninety nursery pigs were used in two metabolism experiments to determine the effects of roasting and extruding on the nutritional value of Williams 82 soybeans with (+K) and without (-K) gene expression for the Kunitz trypsin inhibitor. Treatments for both experiments were: 1) soybean meal; 2) +K roasted; 3) +K extruded; 4) -K roasted; and 5) -K extruded. Diets were the soybean preparations (96.5% of the diet) with only vitamins and minerals added as needed to meet or exceed NRC recommendations. Daily feed allowance was 5% of initial BW given as three equal meals. In Exp. 1, 50 weanling pigs (4.7 kg average BW and 21 d average age) were used. Apparent values for N digestibility (p<0.001), biological value (p<0.09), percentage N retention (p<0.05), GE digestibility (p<0.001) and percentage ME (p<0.001) were greater for pigs fed extruded soybeans than pigs fed roasted soybeans. Also, N digestibility (p<0.05), biological value (p<0.03) and percentage N retention (p<0.04) were greater for pigs fed -K soybeans than those fed +K soybeans. In Exp. 2, 40 pigs (9.7 kg average BW and 35 d average age) were allowed to adjust to the nursery environment before use in the experiment. In general, the pigs in Exp. 2 (i.e., the older pigs) had greater utilization of nutrients from all of the soybean products than the younger pigs used in Exp. 1. Digestibilities of DM, N and GE were greater (p<0.001) for pigs fed -K soybeans than those fed +K soybeans and extruded soybeans had greater digestibilities of DM, N and GE than roasted soybeans (p<0.001). Also, percentage N retention (p<0.01) and percentage ME (p<0.001) for pigs fed extruded soybeans were greater than for pigs fed roasted soybeans. In conclusion, extruded and -K soybeans were greater nutritional value than roasted and +K soybeans for 4.7 and 9.7 kg nursery pigs.

Effects of Ulinastatin on Postoperative Blood Loss and Hemostasis in Atrioventricular Valve Surgery with Cardiopulmonary Bypass

  • Park, Jae Bum;Kim, Seong Hyop;Lee, Song Am;Chung, Jin Woo;Kim, Jun Seok;Chee, Hyun Keun
    • Journal of Chest Surgery
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    • v.46 no.3
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    • pp.185-191
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    • 2013
  • Background: Cardiopulmonary bypass (CPB) induces variable systemic inflammatory reactions associated with major organ dysfunction via polymorphonuclear neutrophils (PMNs). Ulinastatin, a urinary trypsin inhibitor, inhibits PMN activity and reduces systemic inflammatory responses. The aim of this study is to evaluate the effect of ulinastatin on postoperative blood loss and laboratory changes in patients undergoing open heart surgery. Materials and Methods: Between January 2008 and February 2009, 110 patients who underwent atrioventricular valve surgery through right thoracotomy were divided into two groups. Patients received either 5,000 U/kg ulinastatin (ulinastatin group, n=41) or the equivalent volume of normal saline (control group, n=69) before aortic cross clamping. The primary end points were early coagulation profile changes, postoperative blood loss, transfusion requirements, and duration of intubation and intensive care unit stay. Results: There were no statistically significant differences between the two groups in early coagulation profile, other perioperative laboratory data, and postoperative blood loss with transfusion requirements. Conclusion: Administration of ulinastatin during operation did not improve the early coagulation profile, postoperative blood loss, or transfusion requirements of patients undergoing open heart surgery. In addition, no significant effect of ulinastatin was observed in major organs dysfunction, systemic inflammatory reactions, or other postoperative profiles.

Ovalbumin Hydrolysates Inhibit Nitric Oxide Production in LPS-induced RAW 264.7 Macrophages

  • Kim, Hyun Suk;Lee, Jae Hoon;Moon, Sun Hee;Ahn, Dong Uk;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.40 no.2
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    • pp.274-285
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    • 2020
  • In this study, ovalbumin (OVA) hydrolysates were prepared using various proteolytic enzymes and the anti-inflammatory activities of the hydrolysates were determined. Also, the potential application of OVA as a functional food material was discussed. The effect of OVA hydrolysates on the inhibition of nitric oxide (NO) production was evaluated via the Griess reaction, and their effects on the expression of inducible NO synthase (inducible nitric oxide synthase, iNOS) were assessed using the quantitative real-time PCR and Western blotting. To determine the mechanism by which OVA hydrolysates activate macrophages, pathways associated with the mitogen-activated protein kinase (MAPK) signaling were evaluated. When the OVA hydrolysates were added to RAW 264.7 cells without lipopolysaccharide (LPS) stimulation, they did not affect the production of NO. However, both the OVA-Protex 6L hydrolysate (OHPT) and OVA-trypsin hydrolysate (OHT) inhibited NO production dose-dependently in LPS-stimulated RAW 264.7 cells. Especially, OHT showed a strong NO-inhibitory activity (62.35% at 2 mg/mL) and suppressed iNOS production and the mRNA expression for iNOS (p<0.05). Also, OHT treatment decreased the phosphorylation levels of Jun amino-terminal kinases (JNK) and extracellular signal-regulated kinases (ERK) in the MAPK signaling pathway. These findings suggested that OVA hydrolysates could be used as an anti-inflammatory agent that prevent the overproduction of NO.

Effects of Dietary Fructooligosaccharide on Digestive Enzyme Activities, Intestinal Microflora and Morphology of Growing Pigs

  • Xu, Z.R.;Zou, X.T.;Hu, C.H.;Xia, M.S.;Zhan, X.A.;Wang, M.Q.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.12
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    • pp.1784-1789
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    • 2002
  • One hundred and twenty-eight growing barrows (Jiaxing Black${\times}$Duroc${\times}$Landrace) at an average BW of 20.8 kg were allocated to four treatments for 42 days, each of which was replicated four times with eight pigs per replicate and used to investigate the effects of fructooligosaccharide (FOS) on digestive enzyme activities, intestinal microflora and morphology of growing pigs. The pigs received the same basal corn-soybean meal diet and FOS was added to the basal diet at 0, 2, 4, 6 g/kg diet at the expense of corn, respectively. As compared to control, supplementation with 4 and 6 g/kg FOS significantly improved average daily gain and feed efficiency. Addition of FOS enhanced the growth of Bifidobacterium and Lactobacillus, but inhibited Clostridium and Escherichia coli in the small intestinal and proximal colonic contents. Supplementation with 4 and 6 g/kg FOS significantly improved the activities of total protease, trypsin and amylase in the small intestinal contents. However, FOS had no significant effect on the activity of lipase in the small intestinal contents as well as the digestive enzymes in pancreas. Morphological measurement of jejunal mucosa did show response to consumption of FOS. Villus height and the villus height to crypt depth ratio at the jejunal mucosa were significantly higher with 4 and 6 g/kg FOS supplementation as compared to control.

Effect of First Line Gastric Cancer Chemotherapy Regime on the AGS Cell Line - MTT Assay Results

  • Alizadeh-Navaei, Reza;Rafiei, Alireza;Abedian-Kenari, Saeid;Asgarian-Omran, Hossein;Valadan, Reza;Hedayatizadeh-Omran, Akbar
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.1
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    • pp.131-133
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    • 2016
  • Background: Combination chemotherapy regimes are common treatments for cancer. The aim of this study was to evaluation the effect of individual chemotherapeutic agents in comparison with a first line chemotherapy regime treatment in the AGS gastric cancer cell line by MTT assay. Materials and Methods: In this experimental study, AGS cells were grown in RPMI-1640 supplemented with 10% fetal calf serum and 100 IU/ml penicillin, and $10{\mu}g/ml$ streptomycinin, under a humidified condition at $37^{\circ}C$ with 5% CO2. All cells were washed with PBS and detached with trypsin, centrifuged and 8000 cells re-plated on to 96- well plates. LD50 doses of Epirubicin, Cisplatin and 5-fluorouracil were added to each well in mono or triple therapy. Anti-proliferative activities were determined by MTT assay after 24, 48 or 72 h. Results: Results of MTT assays showed that there were no significant differences among 3 drugs in monotherapy (p=0.088), but there was significant difference between combination therapy with epirubicin (P=0.031) and 5FU (p=0.013) on cell survival at 24 h. After 48 and 72 hours, cell viability showed significant differences between the 3 drugs (p=0.048 and P=0.000 for 48 and 72 h, respectively) and there was significant difference between combination therapy with epirubicin (P=0.035 and P=0.002 for 48 and 72 h, respectively). Conclusions: The results showed no significant differences between these chemotherapy drugs each given alone, but combination therapy with 3 drugs had significant effects on cell viability in comparison with epirubicin alone.

Expression and Biochemical Characterization of the Bacillus thuringiensis Cry4B ${\alpha}1$-${\alpha}5$ Pore-forming Fragment

  • Puntheeranurak, Theeraporn;Leetacheewa, Somphob;Katzenmeier, Gerd;Krittanai, Chartchai;Panyim, Sakol;Angsuthanasombat, Chanan
    • BMB Reports
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    • v.34 no.4
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    • pp.293-298
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    • 2001
  • Tryptic activation of the 130-kDa Bacillus thuringiensis Cry4B $\delta$-endotoxin produced protease-resistant products of ca. 47 kDa and ca. 21 kDa. The 21-kDa fragment was identified as the N-terminal five-helix bundle (${\alpha}1-{\alpha}5$,) which is a potential candidate for membrane insertion and pore formation. In this study, we constructed the recombinant clone over-expressing this putative pore-forming (PPF) fragment as inclusion bodies in Escherichia coli. The partially purified inclusions were composed of a 23-kDa protein, which cross-reacted with Cry4B antibodies, and whose N-terminus was identical to that of the 130-kDa protein. Dissimilar to protoxin inclusions, the PPF inclusions were only soluble when the carbonate buffer, pH 9.0, was supplemented with 6 M urea. After renaturation via a stepwise dialysis, the refolded PPF protein appeared to exist as an oligomer and was structurally stable upon trypsin treatment. Unlike the 130kDa protoxin, the refolded protein was able to release entrapped glucose from liposomes, and showed comparable activity to the full-length activated toxin, although it lacks larvicidal activity These results, therefore, support the notion that the PPF fragment that consists of ${\alpha}1-{\alpha}5$ of the activated Cry4B toxin is involved in membrane pore-formation.

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Identification of Essential Histidines in Cyclodextrin Glycosyltransferase Isoform 1 from Paenibacillus sp. A11

  • Kaulpiboon, Jarunee;Pongsawasdi, Piamsook
    • BMB Reports
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    • v.36 no.4
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    • pp.409-416
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    • 2003
  • The isoform 1 of cyclodextrin glycosyltransferase (CGTase, EC 2.4.1.19) from Paenibacillus sp. A11 was purified by a preparative gel electrophoresis. The importance of histidine, tryptophan, tyrosine, and carboxylic amino acids for isoform 1 activity is suggested by the modification of the isoform 1 with various group-specific reagents. Activity loss, when incubated with diethylpyrocarbonate (DEP), a histidine modifying reagent, could be protected by adding 25 mM methyl-$\beta$-cyclodextrin substrate prior to the modification. Inactivation kinetics of isoform 1 with DEP resulted in second-order rate constants ($k_{inactivation}$) of $29.5\;M^{-1}s^{-1}$. The specificity of the DEP-modified reaction for the histidine residue was shown by the correlation between the loss of isoform activity and the increase in the absorbance at 246 nm of N-carbethoxyhistidine. The number of histidines that were modified by DEP in the absence and presence of a protective substrate was estimated from the increase in the absorbance using a specific extinction coefficient of N-carbethoxyhistidine of $3,200\;M^{-1}cm^{-1}$. It was discovered that methyl-$\beta$-CD protected per mole of isoform 1, two histidine residues from the modification by DEP. To localize essential histidines, the native, the DEP-modified, and the protected forms of isoform 1 were digested by trypsin. The resulting peptides were separated by HPLC. The peptides of interest were those with $R_t$ 11.34 and 40.93 min. The molecular masses of the two peptides were 5,732 and 2,540 daltons, respectively. When the data from the peptide analysis were checked with the sequence of CGTase, then His-140 and His-327 were identified as essential histidines in the active site of isoform 1.