• Title/Summary/Keyword: trypsin.

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Enzymatic Desugarization of Egg White for Drying with Glucose Oxidase (Glucose Oxidase에 의(依)한 건조용(乾燥用) 난백(卵白)의 효소적(酵素的) 탈당(脫糖))

  • Song, Kwang Taek;Oh, Hong Rock;Kwon, Soon Ki;Lee, Bong Duck
    • Korean Journal of Agricultural Science
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    • v.11 no.2
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    • pp.223-232
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    • 1984
  • The influences of some factors involved in removing glucose from egg white by the glucose oxidase system be fore drying were investigated. And the properties between foams prepared from raw and enzyme-treat ed egg white was compared. The results obtained we re summarized as follows; 1. The dianisidine method was found to be suitable for the measurement of egg white glucose in the range up to 100ug/ml. 2. The optimal pH of glucose oxidase activity on glucose was found to be a bout 5.0, and thats activity was most stable in the pH range of about 4.0~5.0 when that enzyme was treat ed for 30 minute at $50^{\circ}C$. 3. The optimal temperature for glucose oxidase reaction on glucose was found to be about $20^{\circ}C$, and that enzyme activity was s table up to $50^{\circ}C$. 4. The removing rate of glucose from egg white with glucose oxidase was influenced by the enzyme concentration, pH and oxygen addition, and the react ion time of the desugarization was about 10 hour sunder the conditions of 0.5% hydrogen peroxide, pH 7.0 and $26^{\circ}C$. 5. All of the each egg white treated with glucose oxidase, glucose oxidase+pancreatin, glucose oxidase+trypsin showed highly foaming ability than that of natural egg white(control), but thats foam stability, on the contrary, was reversed.

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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).

Effect of Environmental Factors on the Growth of Rabbit Oral Keratinocytes (토끼 구강점막 상피세포 성장에 미치는 환경인자의 영향)

  • Yoon, Moon-Young;Park, Hee-Jung;Lee, Doo-Hoon;Jang, In-Keun;Park, Jung-Keug;Kim, Woo-Sik
    • Korean Chemical Engineering Research
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    • v.43 no.1
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    • pp.103-109
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    • 2005
  • Isolation and primary culture technique of rabbit oral keratinocytes, and the study for effect of environmental factors on the cell growth were carried out in T75-flask. $1.92{\pm}0.59{\times}10^6$ viable cells were isolated by trypsin enzymatic digestion method from $0.25cm^2$ biopsy of rabbit oral mucosa. Primary culture with 10 mL of K-SFM containing 50 mg/L BPE, $5.0{\mu}g/L$ EGF and 0.15 mM $Ca^{2+}$ showed confluence after 8 days and doubling time was 2.54 days. Effect of medium types, medium volume and supplement types on the cell growth was investigated after the cultured keratinocytes had been harvested from primary confluence. Serum addition showed adverse effect and the increase of serum concentration didn't have an effect on the cell growth. The increase of medium volume decreased the cell growth. The increase of calcium concentration increased the cell growth and 2.0 mM was optimum value. In conclusion, when rabbit oral keratinocytes was cultured in T75-flask, the most effective conditions was to use 10 mL of K-SFM containing 50 mg/L BPE, $5.0{\mu}g/L$ EGF and 2.0 mM $Ca^{2+}$, and doubling time was 1.32 days. This study can provide the useful informations to develop a process and design a bioreactor for the culture of keratinocytes in human body like skin and cornea, as well as mucosa.

Analyses of Nutrients and Antinutrients of Rice Cultivars (국내 주요 쌀 품종별 영양성분 및 주요 항영양인자 분석)

  • Kyoun, Oh-Yun;Oh, Sang-Hee;Kim, Hyun-Ju;Lee, Jeong-Hee;Kim, Hyoung-Chin;Yoon, Won-Kee;Kim, Hwan-Mook;Kim, Mee-Ree
    • Korean journal of food and cookery science
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    • v.22 no.6 s.96
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    • pp.949-956
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    • 2006
  • Eleven rice varieties, five normal-yielding and six high-yielding, were examined for proximate composition, mineral and fatty acid composition, and antinutrients. The proximate rice compositions for moisture, crude protein, crude fat, carbohydrate, crude fiber and crude ash were 10.70${\sim}$15.88%, 5.03${\sim}$7.73%, 0.45${\sim}$1.33%, 74.56${\sim}$82.05%, 0.30${\sim}$0.73% and 0.31${\sim}$0.91%, respectively. The proximate composition of the various rice varieties was not significantly different except for moisture and carbohydrate. The contents of saturated fatty acid, monounsaturated fatty acid and polyunsaturated fatty acid of the rice varieties were 22.45${\sim}$25.99%, 36.50${\sim}$44.99% and 32.01${\sim}$46.02%, respectively. Several high-yielding rices varieties had higher oleic acid or linoleic acid. Ilmi and Nampyung had high oleic acid contents at ever 40%, and Dongjin No. 1, IlPum, Junam and Seachuchung had high linoleic acid contents at over 37%. The major minerals of rice were K and Mg and their respective contents were 85.62% and 25.55%. High-yielding rice varieties had significantly higher Mn and Na contents and lower Mg and Fe contents than normal-yielding rice varieties (p<0.05). Antinutrients such as lectin, phytic acid and trypsin inhibitor were not detected in any of the rice varieties.

Antigenicity of Whey Protein Hydrolysates against Rabbit Anti ${\beta}-Lactoglobulin$ Antiserum (토끼 항 ${\beta}-Lactoglobulin$ 항혈청에 대한 유청단백질 가수분해물의 항원성)

  • Lee, Soo-Won;Ha, Woel-Kyu;Juhn, Suk-Lak;Kim, Jung-Wan;Shon, Dong-Hwa;Lee, Jae-Young
    • Korean Journal of Food Science and Technology
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    • v.26 no.5
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    • pp.532-538
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    • 1994
  • In order to investigate the lowering effects of in vitro enzymatic hydrolysis by the treatment of chymotrypsin, trypsin, pancreatin, or protease from Aspergillus oryzae on the antigenicity of whey protein(WPI) against rabbit anti ${\beta}-LG$ antiserum, competitive inhibition ELISA(cELISA) and passive cutaneous anaphylaxis(PCA) test using guinea pig were performed. The results of cELISA showed that the monovalent antigenicity of the whey protein hydrolysates(WPH) to the antiserum was decreased to $10^{-1.7}{\sim}10^{-4.1}$ and less by the hydrolysis. Especially, the antigenicity of OUP(hydrolysate by protease from Asp. oryzae with preteatment of pepsin) was found almost to be removed. By the heterologous PCA the polyvalent antigenicity of the WPH was decreased to $1/2{\sim}1/128$ and less. Especially, the polyvalent antigenicity of OUN(hydrolysate by protease from Asp. oryzae without preteatments) was found almost to be removed, although OUN did not have so high degree of hydrolysis(DH) or so low monovalent antigenicity (reduced to $10^{-3.2}$). Therefore, this result was assumed to come from effective destruction of antigenic determinants on ${\beta}-LG$ in WPI, not to produce polyvalent antigenic peptides that are closely associated with induction of allergy. This finding suggested that WPH prepared by the treatment of microorganic protease from Asp. oryzae would be a material for hypoallergenic infant formula due to the removal of the polyvalent antigenicity of ${\beta}-LG$, the major milk allergen in WPI.

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The Proteinase Distributed in the Intestinal Organs of Fish 2. Characterization of the Three Alkaline Proteinases from the Pyloric Caeca of Mackerel, Scomber japonicus (어류의 장기조직에 분포하는 단백질분해효소에 관한 연구 2. 고등어 유문수조직중에 분포하는 3종 알칼리성 단백질분해효소의 특성)

  • KIM Hyeung-Rak;PYEUN Jae-Hyeung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.19 no.6
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    • pp.547-557
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    • 1986
  • The characteristics of the three alkaline proteinases, Enz. A, B and C, from the pyloric caeca of mackerel have been investigated. The optimum condition for the activity of the Enz. A, B and C was pH 9.4, 9.8 and 9.8 at $45^{\circ}C$ for $2\%$ casein solution, and was pH 9.2 10.2 and 9.8 at $45^{\circ}C$ for $5\%$ hemoglobin denatured by urea, respectively. Enz. A, B and C by heat treatment at $50^{\circ}C$ for 5 min were inactivated 90, 33 and $37\%$, respectively, over the original activity. The reaction rate of the three alkaline proteinases was constant to the reaction time to 40 min in the reaction condition of $2{\mu}g/ml$ of enzyme concentration and $2\%$ casein solution. The reaction rate equation and Km value against casein substrate determined by the method of Lineweaver and Burk were: Enz. A, Y=3.6X and $Km=5.0{\times}10^{-3}\%$; Enz. B, Y=6.0X and $Km=1.0{\times}10^{-3}\%$; Enz. C, Y=4.2X and $Km=3.6{\times}10^{-3}\%$. The three alkaline proteinases were inactivated by $Ag^+$ and $Hg^{2+}$, but activated by $Mn^{2+},\;Sn^{2+}\;and\;Pb^{2+}$, Enz. B and C were remarkably inhibited by the soybean trypsin inhibitor. Molecular weight of Enz. A, B and C determined by SDS-polyacrylamide gel electrophoresis and Sephadex G-100 gel filtration was in the range of $27,500{\pm}2,500,\;20,500{\pm}1,500\;and\;15,250{\pm}250$, respectively.

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Effect of Blue Light on the Major Components of Soybean-sprouts (Blue광(光) 조사(照射)가 콩나물의 주요성분(主要成分)에 미치는 영향(影響))

  • Kim, Kwang-Soo;Kim, Soon-Dong;Kim, Jin-Koo;Kim, Ju-Nam;Kim, Kyoung-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.11 no.4
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    • pp.7-12
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    • 1982
  • Growth of soybean sprouts(Glycine Max L.) and amounts of some chemical components were measured when they were exposed to blue light (120lux, 3hrs/day) during their growth. Hypocotyl length of irradiated soybean sprouts exceeded slightly that of control (dark) soybean sprouts, but the tfresh weight of whole sprouts as well as each part of the sprout showed no difference between the two groups. Chlorophyll content of cotyledon under blue light increased significantly with the lapse of days (3.57 and $8.45\;{\mu}g/100g$ fresh weight on the 3rd and 7th day). Bluelight irradiated sprouts contained more vitamin C than control sprouts (21.7% and 30.8% higher for the cotyledon and hypocotyl). Total amount of protein was not affected. Hypocotyl protein content was 8% of that in original soybean. Blue light did not affect the activity of trypsin inhibitor of sprouts. Similar activity of the inhibitor was observed in the cotyledon whereas hypocotyl showed activity corresponding to 23.7% of original bean. Polyacrylamide gel electrophoretogram for the protein showed 10, 9, and 11 bands in the original bean. 5th day cotyledon and hypocotyl respectively. Especially, band 3 of low Rm value was major protein component for the hypocotyl. Band 5 and 11 could be seen only in the protein of hypocotyl from bluelight irradiated sprouts, whereas no effect of blue light on the electrophoretogram was observed for the cotyledon.

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Role of HOXA Gene in Human Endometrial Decidualization (인간 자궁내막의 탈락막화에서 HOXA10 유전자의 역할)

  • Lee, Chang-Se;Park, Dong-Wook;Park, Chan-Woo;Kim, Tae-Jin
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.3
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    • pp.207-216
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    • 2010
  • Objective: This study was performed to clarify the role of HomeoboxA (HOXA) and its related signaling molecules in the decidualization of primary cultured endometrial cells. Methods: Human endometrial tissues were obtained by curettage of hysterectomy specimens from patients with conditions other than endometrial diseases. Tissues were minced and digested with Trypsin-EDTA for 20 min, $37^{\circ}C$. Cells were cultured with DMEM/F12 medium in $37^{\circ}C$, 5% $CO_2$ incubator for 24 hrs. Cells were treated with HOXA10 siRNA and added transforming growth factor (TGF)-${\beta}1$ (10 ng/mL) for 48 hrs to induces decidualization in vitro. Reverse transcription polymerase chain reaction analysis was accomplished to observe the expression of HOXA10, prolactin, cyclooxygenase (COX)-2, peroxisome proliferatoractivated receptor (PPAR)-$\gamma$, and wingless-type MMTV integration site family (Wnt). Results: HOXA10 expression was increased (1.8 fold vs. non-treated control) in TGF-${\beta}1$ treated cells. Decidualization marker, prolactin, was significantly increased in TGF-${\beta}1$ treated cells compared with HOXA10 siRNA treated cells. Endometrial cell differentiation marker, COX-2 was down-regulated by HOXA10 siRNA even if cells were treated with TGF-${\beta}1$. Wnt4 was down-regulated by treated with HOXA10 siRNA, this expression patters was not changed by TGF-${\beta}1$. Expression of PPAR-$\gamma$ was down regulated by TGF-${\beta}1$ in regardless of HOXA10 siRNA treatment. Conclusion: TGF-${\beta}1$ which is induced by progesterone in endometrial epithelial cells may induces stromal cell decidualization via HOXA10 and Wnt signaling cascade.

Effects of Dietary Zinc Supplements on Growth, Feed Efficiency, Organ Weight, Blood Biochemical Profiles, and Activity of Digestive Enzymes in Growing Korean Native Chicks (아연 보충급여에 따른 한국재래계의 성장, 사료이용성, 장기무게, 혈액생화학적 성상, 장기무게 및 소화효소 활성도에 미치는 영향)

  • Jeon, Dong-Gyung;Kim, Min-Jeong;Yoon, Il-Gyu;Ahn, Ho-Sung;Moon, Eun-Seo;Sohn, Sea-Hwan;Lim, Yong;Jang, In-Surk
    • Korean Journal of Poultry Science
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    • v.46 no.2
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    • pp.117-125
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    • 2019
  • The aim of the study was to investigate the effects of dietary supplementation of zinc (Zn) sources (zinc oxide and Zn-methionine) on performance, organ weights, blood biochemical profiles, and digestive enzymes of the pancreas and small intestine in Korean native chicks (KNC). A total of 144 KNC (n=6) were fed a basal diet (CON, 100 ppm of Zn), a basal diet supplemented with 50 ppm of Zn with ZnO (ZNO), or a basal diet supplemented with 50 ppm of Zn with Zn-methionine (ZMT) for 28 days. There was no significant difference in body weight, gain, feed intake, and feed conversion ratio among the three groups. The relative weights of the liver, spleen, and intestinal mucosa were unaffected by the dietary source of Zn, whereas pancreas weight in the ZNO group decreased (P<0.05) compared with that in the CON and ZMT groups. Blood biochemical components including aspartate aminotransferase, and alanine aminotransferase were unaffected by dietary Zn supplementation. Pancreatic trypsin activity in the ZNO and ZMT groups was significantly (P<0.05) enhanced compared with that in the CON group. However, the activities of ${\alpha}$-amylase and carboxypeptidase A were not altered by dietary Zn supplementation. The activities of maltase and sucrase were unchanged, whereas the activity of leucine aminopeptidase tended (P=0.08) to be increased by dietary Zn supplementation. In conclusion, the supplementation with 50 ppm of ZnO or Zn-methionine resulted in an activation of protein digestive enzymes in the pancreas and small intestine without affecting animal performance in KNC.

Effects of Recipient Oocytes and Electric Stimulation Condition on In Vitro Development of Cloned Embryos after Interspecies Nuclear Transfer with Caprine Somatic Cell (수핵난자와 전기적 융합조건이 산양의 이종간 복제수정란의 체외발달에 미치는 영향)

  • 이명열;박희성
    • Reproductive and Developmental Biology
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    • v.28 no.1
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    • pp.21-27
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    • 2004
  • This study was conducted to investigate the developmental ability of caprine embryos after somatic cell interspecies nuclear transfer. Recipient bovine and porcine oocytes were obtained from slaughterhouse and were matured in vitro according to established protocols. Donor cells were obtained from an ear-skin biopsy of a caprine, digested with 0.25% trypsin-EDTA in PBS and primary fibroblast cultures were established in TCM-199 with 10% FBS. The matured oocytes were dipped in D-PBS plus 10% FBS + 7.5 $\mu$ g/ml cytochalasin B and 0.05M sucrose. Enucleation were accomplished by aspirating the first polar body and partial cytoplasm which containing metaphase II chromosomes using a micropipette with an out diameter of 20∼30 $\mu$m. A Single donor cell was individually transferred into the perivitelline space of each enucleated oocyte. The reconstructed oocytes were electric fusion with 0.3M mannitol fusion medium. After the electrofusion, embryos were activated by electric stimulation. Interspecies nuclear transfer embryos with bovine cytoplasts were cultured in TCM-199 medium supplemented with 10% FBS including bovine oviduct epithelial cells for 7∼9 day. And porcine cytoplasts were cultured in NCSU-23 medium supplemented with 10% FBS for 6 ∼8 day at $39^{\circ}C, 5% CO_2 $in air. Interspecies nuclear transfer by recipient bovine oocytes were fused with electric length 1.95 kv/cm and 2.10 kv/cm. There was no significant difference between two electric length in fusion rate(47.7 and 44.6%) and in cleavage rate(41.9 and 54.5%). Using electric length 1.95 kv/cm and 2.10 kv/cm in caprine-porcine NT oocytes, there was also no significant difference between two treatments in fusion rate(51.3 and 46.1%) and in cleavage rate(75.0 and 84.9%). The caprine-bovine NT oocytes fusion rate was lower(P<0.05) in 1 pulse for 60 $\mu$sec(19.3%), than those from 1 pulse for 30 $\mu$sec(50.8%) and 2 pulse for 30 $\mu$sec(31.0%). The cleavage rate was higher(P<0.05) in 1 pulse for 30 $\mu$sec(53.3%) and 2 pulse for 30 $\mu$sec(50.0%), than in 1 pulse for 60 $\mu$sec(18.2%). The caprine-porcine NT oocytes fusion rate was 48.1% in 1 pulse for 30 $\mu$sec, 45.2% in 2 pulse for 30 $\mu$sec and 48.6% in 1 pulse for 60 $\mu$sec. The cleavage rate was higher(P<0.05) in 1 pulse for 30 $\mu$sec(78.4%) and 1 pulse for 60 $\mu$sec(79.4%), than in 2 pulse for 30 $\mu$sec(53.6%). In caprine-bovine NT embryos, the developmental rate of morula and blastocyst stage embryos were 22.6% in interspecies nuclear transfer and 30.6% in parthenotes, which was no significant differed. The developmental rate of morula and blastocyst stage embryos with caprine-porcine NT embryos were lower(P<0.05) in interspecies nuclear transfer(5.1%) than parthenotes(37.4%).