• Title/Summary/Keyword: Protease digestion

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Effect of Meju Shapes and Strains on the Quality of Soy Sauce (Aspergillus oryzae 및 Aspergillus sojae를 이용한 개량(改良)메주의 형상에 의한 장류(醬類)의 품질비교)

  • Kim, Sang-Soon
    • Korean Journal of Food Science and Technology
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    • v.10 no.1
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    • pp.63-72
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    • 1978
  • Effect of shapes (noodle, grain and brick types) and strains (Aspergillus oryzae and Aspergillus sojae) of Meju (microorganism inoculated soybean substrate for fermentation) on the quality of soy sauce was investigated. Generally, the highest protease activity was found in the noodle type-Meju inoculated Asp. sojae during Meju preparation and soy sauce brewing, however, the lowest value was noted in brick type. Meju inoculated Asp. oryzae. Similar tendency was found on the contents of total solid, total nitrogen, amino-nitrogen, nitrogen digestion yield and amino-nitrogen ratio during soy sauce brewing. No effect was shown on the reducing sugar content, alcohol formation, pH, buffer action and salt content according to different Meju types during soy sauce brewing. Organoleptic tests on the color, taste and flavor of soy sauce after 3 months brewing with various Meju scored in the order of noodle type-Asp. sojae soy sauce (best quality), noodle type-Asp, orzae soy sauce, brick type-Asp. sojae soy sauce and brick type-Asp. oryzae-soy sauce (worst quality).

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Processing of Enzymatic Hydrolysates from Conger eel Scrap (붕장어 가공잔사를 이용한 효소분해소재의 가공)

  • Kang Su Tae;Kong Chung Sik;Cha Yong Jun;Kim Jong Tae;Oh Kwang Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.3
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    • pp.259-264
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    • 2002
  • In order to develope nutritional and flavoring intermediate products, the optimal processing conditions for two stage enzyme hydrolysate (TSEH) from low-utilized conger eel scrap such as head and intestine were investigated. The optimal processing conditions for TSEH were revealed in temperature at $55^{\circ}C$ 3$\~$4 hours digestion with alcalase at the 1st stage, and 4 hours at $45{\~}50^{\circ}C$ digestion with neutrase at the 2nd stage. Among water extract, steam extract and enzyme hydrolysates of conger eel scrap, the present TSEH was superior to other extracts in terms of yield ana organoleptic taste such as harmonic umami and inhibition of fishy and greasy taste formation. From the results of chemical experiments and sensory evaluation, we may conclude that TSEH of conger eel scrap could be utilized as the flavoring intermediate materials for the fisheries products such as flavoring sauces, drinkable beverage and instant food materials.

Electrophoretic Patterns of Myofibrillar Proteins by Sugar Addition and Heat Treatment (당첨가 및 가열처리에 의한 근원섬유 단백질의 전기영동 Pattern 변화)

  • Yang, Jong-Beom;Yoon, Won-Ho;Ko, Myung-Soo;Kim, Chang-Han
    • Korean Journal of Food Science and Technology
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    • v.22 no.6
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    • pp.640-645
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    • 1990
  • Changes of the electrophoretic patterns of myofibrillar proteins by sugar audition and heat treatment was studied. In the electrophoretic patterns of myofibrills prepared from no sugar added meat, as the intensity of higher molecular weight band such as myosin heavy chain showed a remarkable decrease by heating, that of lower molecular weight band such as actin showed no change. That from sugar added meat showed more remarkable decrease in the intensity of higher molecular weight band than that from no sugar added meat and this tendency was most noticeable in case of glucose addition. The effect of digestion with proteases after sugar addition and heat treatment on the electrophoretic patterns exhibited the descending order of trypsin >chymotrypsin >peptidase. By digestion with these three enzymes at one time myosin produced 27.000 dalton and 32.000 dalton components, and actin showed 16,000 dalton component. in the case of heat treatment, a part of actin was not digested. And in the case of glucose addition the myosin aggregates was not digested with these three enzymes at a time.

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Effects of Capsosiphon fulvescens Extracts on Essential Amino Acids Absorption in Rats (매생이 열수추출물이 흰쥐의 필수아미노산 흡수에 미치는 영향)

  • Kim, Hyo-Young;Kim, In-Hye;Nam, Taek-Jeong
    • Journal of Life Science
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    • v.19 no.11
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    • pp.1591-1597
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    • 2009
  • The aim of this study was to examine the effects of Capsosiphon fulvescens extract (CFE) on levels of essential amino acids in serum and the rate of protein digestion. CFE contains approximately 68% carbohydrate as fiber, 17% crude ash, and 14% crude protein. Experimental rats were divided two groups, one group receiving a normal diet and the other a normal diet containing 5% CFE. To analyze the absorption of essential amino acids into the hepatic portal vein, blood from the hepatic portal vein was collected between 10 a.m. and 5 p.m. for 10 days. In vitro protein digestion was decreased in the group fed 5% CFE; these data indicated that CFE affected protease activity. We measured the absorption of essential amino acids in the serum of the hepatic portal vein, at 30-, 60-, 90-, and 120 minutes after feeding. Although there was no difference in the concentration of total essential amino acids between the two groups, the 5% CFE-fed animals had a decreased rate of absorption. Absorption of Lys and Thr into the hepatic portal vein was lower in the CFE-fed group than in the control group. The rate of absorption of Met was delayed nearly 50% in the CFE-fed group compared to the control group. On the other hand, the rate of absorption of Leu, Ile, and Val was increased; Phe showed the same. Therefore, we suggest that CFE could affect protein metabolism by increasing or decreasing the absorption rate of essential amino acids.

Total Folate Contents of 15 Edible Plants Consumed in Korea Using Trienzyme Extraction Method (국내 소비되는 봄나물의 Trienzyme 추출법을 적용한 엽산 함량 분석)

  • Kim, Bo Min;Kim, So-Min;Oh, Ji Yeon;Cho, Young-Sook;Kim, Se-Na;Choi, Youngmin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.11
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    • pp.1796-1800
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    • 2014
  • Trienzyme digestion (AOAC Official Method 2004.05) procedure using protease, ${\alpha}$-amylase, and chicken pancreas conjugase was evaluated to determine its usefulness in the microbiological quantitation of total folate in foods. Folate values obtained by alkali hydrolysis (Korean Food Standards Codex) were compared to those obtained by the trienzyme method for four certified reference materials (CRM) representing diverse matrixes. Trienzyme treatment increased measurable folate from most CRM compared to levels found after alkali hydrolysis. The largest increases were observed with CRM 487 (pig liver, 5.8-fold) and CRM 121 (whole meal flour, 3.1-fold) after trienzyme digestion. Using trienzyme digestion method, total folate contents of raw and blanched edible plants were determined. Eleutherococcus senticosus ($146.9{\mu}g/100g$) showed the highest total folate content, followed by Aster glehni F. Schmidt ($142.8{\mu}g/100g$) and Ledebouriella seseloides H. Wolff ($140.4{\mu}g/100g$) on a wet weight basis. Blanching of samples resulted in an insignificant decrease in folate content for five samples and 11~63% reduction for nine samples. Our finding suggests that trienzyme digestion method is accurate for the determination of food folate in leafy vegetables.

Apoptosis of Human Hepatocarcinoma (HepG2) and Neuroblastoma (SK-N-SH) Cells Induced by Polysaccharides-Peptide Complexes Produced by Submerged Mycelial Culture of an Entomopathogenic Fungus Cordyceps sphecocephala

  • Oh, Jung-Young;Baek, Yu-Mi;Kim, Sang-Woo;Hwang, Hye-Jin;Hwang, Hee-Sun;Lee, Sung-Hak;Yun, Jong-Won
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.512-519
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    • 2008
  • Three different polysaccharide-peptide complexes (PPC, named as Fr-I, Fr-II, and Fr-III) were produced by submerged mycelial culture of an entomopathogenic fungus Cordyceps sphecocephala, and their anticancer activities were investigated in human hepatocarcinoma (HepG2) and neuroblastoma (SK-N-SH) cells. The highest inhibitory effects of PPC on both HepG2 and SK-N-SH cells were achieved with Fr-I, whereas Fr-III with low molecular mass showed lower inhibition effects. Interestingly, the inhibitory effects of the three fractions were increased after protease digestion, suggesting that the inhibitory effects resulted mainly from the carbohydrate moiety, at least in the case of Fr-II and Fr-III, of PPC. The results of DNA fragmentation in PPC-induced apoptotic cells were confirmed by both DNA ladder assay and comet assay. Our investigation also showed that PPC-induced apoptosis of both cancer cells was associated with intracellular events including DNA fragmentation, activation of caspase-3, and modulation of Bcl-2 and Bax. We conclude that PPC has potential as a novel therapeutic agent for the treatment of both HepG2 and SK-N-SH cancer cells without any cytotoxicity against normal cells.

Anti-Complementary Properties of Polysaccharides Isolated from Fruit Bodies of Mushroom Pleurotus ostreatus

  • Kweon, Mee-Hyang;Jang, Hyo;Lim, Wang-Jin;Chang, Hyo-Ihl;Kim, Chan-Wha;Yang, Han-Chul;Hwang, Han-Joon;Sung, Ha-Chin
    • Journal of Microbiology and Biotechnology
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    • v.9 no.4
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    • pp.450-456
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    • 1999
  • A high molecular-weight water-soluble fraction(PO) obtained by the ethanol precipitation of 0.1 N NaOH extracts of the mushroom Pleurotus ostreatus showed 82% anti-complementary activity for complement consumption hemolysis. The PO consisted of 42% carbohydrate (w/w), 50% protein (w/w), and 3% uronic acid (w/w). Fifty-eight percent of the anti-complementary activity decreased by periodate oxidation and 22% by protease digestion, suggesting that the sugar and protein moieties are essential for this activity. Two polysaccharide fractions, PO-IIIa-1 and PO-IIIa-2, with anti-complementary activity were isolated from the PO using DEAE-Sepharose FF followed by Sephadex G-75 and Sepharose CL-6B gel permeation chromatographies. The PO-IIIa-2 was found by HPLC to be nearly homogeneous, with the molecular mass of 531 kDa, and showed 96% $ITCH_{50}$ (inhibition against the total complement hemolysis of deionized water as the control) at a concentration of 1 mg/ml. This fraction contained galactose, mannose, fucose, and glucose with molar ratios of 1.75:1:0.65 and 0.59, respectively. The majority of galactose and mannose units in the PO-IIIa-2 were composed of TGalp1 ->, ->6Galp1->, ->2,6Galp1->, and ->Manp1->. The PO-IIIa-1 (molecular mass of 2000 kDa), exhibiting higher activity than the PO-IIIa-2, was further purified into two fractions, unbound proteoglycan (PO-IIIa-1A) and bound glucomannan (PO-IIIa-lB), by affinity chromatography using ConA-Sepharose CL-4B. The anti-complementary activity of each affinity purified fraction decreased as compared to that of the native PO-IIIa-1 fraction, indicating that the formation of complex between both polysaccharide fractions was necessary for full anti-complementary activity.

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Novel Preparation and Characterization of the α4-loop-α5 Membrane-perturbing Peptide from the Bacillus thuringiensis Cry4Ba δ-endotoxin

  • Leetachewa, Somphob;Katzenmeier, Gerd;Angsuthanasombat, Chanan
    • BMB Reports
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    • v.39 no.3
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    • pp.270-277
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    • 2006
  • Helices 4 and 5 of the Bacillus thuringiensis Cry4Ba $\delta$-endotoxin have been shown to be important determinants for mosquito-larvicidal activity, likely being involved in membrane-pore formation. In this study, the Cry4Ba mutant protein containing an additional engineered tryptic cleavage site was used to produce the $\alpha4$-$\alpha5$ hairpin peptide by an efficient alternative strategy. Upon solubilization of toxin inclusions expressed in Escherichia coli and subsequent digestion with trypsin, the 130-kDa mutant protoxin was processed to protease-resistant fragments of ca. 47, 10 and 7 kDa. The 7-kDa fragment was identified as the $\alpha4$-loop-$\alpha5$ hairpin via N-terminal sequencing and mass spectrometry, and was successfully purified by size-exclusion FPLC and reversed-phase HPLC. Using circular dichroism spectroscopy, the 7-kDa peptide was found to exist predominantly as an $\alpha$-helical structure. Membrane perturbation studies by using fluorimetric calcein-release assays revealed that the 7-kDa helical hairpin is highly active against unilamellar liposomes compared with the 65-kDa activated full-length toxin. These results directly support the role of the $\alpha4$-loop-$\alpha5$ hairpin in membrane perturbation and pore formation of the full-length Cry4Ba toxin.

Production of Leptin in E. coli and Its Effect on Glucose and Acetate Transport and Expression of Uncoupling Protein-2 Gene in Adipose Tissues of Korean Cattle (Hanwoo)

  • Kim, K.S.;Baik, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.8
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    • pp.1062-1068
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    • 2004
  • Leptin has a major role in the regulation of food intake and energy homeostasis. In addition, leptin participates in many physiological functions including regulation of lipid metabolism. Bovine recombinant leptin protein was produced in E. coli cells in order to understand function of leptin in the regulation of lipid metabolism. The leptin expression vector was constructed in pGEX-4T-3 vector and transformed into E. coli BL21 cells. Expression of the GST-leptin fusion protein was induced with IPTG. The fusion protein was purified using glutathione sepharose 4B batch method, and the recombinant leptin was eluted after thrombin protease digestion. The effect of leptin on glucose transport was examined in the differentiated adipocytes of 3T3-L1 cells. Leptin had no effect on basal and insulin-stimulated glucose transport in 3T3-L1 cells (p>0.05). Effect of recombinant leptin on glucose and acetate transport was examined in adipose tissues of Korean cattle (Hanwoo). Insulin stimulated glucose transport in both intramuscular and subcutaneous adipose tissues (p<0.05), but leptin did not affect glucose transport in both adipose tissues (p>0.05). Insulin stimulated acetate transport in bovine adipose tissues (p<0.05), but leptin did not affect acetate transport (p>0.05). Northern and RT-PCR analyses showed that mRNA levels of uncoupling protein-2 were increased by leptin treatment in 3T3-L1 cells without statistical difference (p>0.05). In conclusion, bovine recombinant leptin did not affect glucose and acetate transport in both 3T3-L1 adipocytes and bovine adipose tissues, while it stimulates UCP-2 mRNA expression in 3T3-L1 cells.

Effects of Enzyme Supplementation on Growth, Intestinal Content Viscosity, and Digestive Enzyme Activities in Growing Pigs Fed Rough Rice-based Diet

  • Wang, M.Q.;Xu, Z.R.;Sun, J.Y.;Kim, B.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.2
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    • pp.270-276
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    • 2008
  • The purpose of the present study was to investigate the effects of exogenous non-starch polysaccharides (NSP) enzymes on performance, intestinal content viscosity and digestive enzyme activities of growing pigs fed a rough rice-based diet. A total of 60 crossbred barrows with an initial body weight of 35.16 kg (SD = 0.82) were blocked by body weight and randomly assigned to two treatments with three replications. Each group was fed the diet based on rice with or without exogenous NSP enzymes (2 g/kg of diet). During the 70 days of the feeding trial, all pigs were given free access to feed and water. At the end of the feeding trial, six pigs from each treatment were randomly selected and slaughtered to collect intestinal digesta, intestinal mucosa, and pancreas. The addition of NSP enzymes improved average daily gain (p<0.05) and feed:gain (p<0.05), and decreased viscosity of digesta in the jejunum (p<0.001) and ileum (p<0.01) of pigs. The supplementation of NSP enzymes increased activities of protease (p<0.01), trypsin (p<0.01) and ${\alpha}$-amylase (p<0.05) in duodenal contents. However, digestive enzymes in the pancreas, jejunal and ileal mucosa were unaffected by the supplemental NSP enzymes (p>0.10). The results indicate that the addition of NSP enzymes to rough rice-based diets improved performance of pigs, reduced viscosity and increased digestive activity in the small intestine.