• Title/Summary/Keyword: Protease stability

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Application of Temperature Gradient Gel Electrophoresis To cAMP Receptor Protein (온도 기울기 전기영동장치의 CAMP 수용성 단백질에 응용)

  • Gang, Jong-Back;Cho, Hyun-Young
    • Journal of Life Science
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    • v.14 no.2
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    • pp.309-314
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    • 2004
  • Cyclic AMP receptor protein (CRP) is involved in the transcriptional regulation of more than 100 genes in E. coli. CRP dimer is converted into active form via the sequential conformation change of cAMP binding pocket, hinge region and HTH DNA binding motif by binding of cAMP. The temperature gradient gel electrophoresis (TGGE) was applied to CRP protein to know whether it was an efficient technique to study the conformational transitions and the thermal stability. TGGE showed the unfolding process of wild-type and S83G CRP proteins with the temperature gradient set from 29 to 71$^{\circ}C$ on nondenaturing polyacrylamide gel. Melting temperature (Tm) was 57$\pm$1 and 55$\pm$1$^{\circ}C$ for wild-type and S83G CRP, respectively in acidic buffer[89.8 mM Glycine and 24 mM Boric acid (pH 5.8)].

Processing of Squeezed-Type Cockle Shell By-Product Paste (새조개 처리동결 가공부산물을 이용한 페이스트 젓갈 제조)

  • 배태진;강훈이
    • The Korean Journal of Food And Nutrition
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    • v.10 no.4
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    • pp.528-532
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    • 1997
  • A fermented squeezed-type paste was processed in order to highly effective utilization of cockle shell by-product, and improvement on rheological properties and texture of hydrolysate by used additives. The cockly shell by-products were homogenized with addition of water and enzymatically hydrolyzed at 5$0^{\circ}C$ for 8 hours added 4% Protease N.P.(Pacific Chemical Co.). And the hydrolysate was thermally treated for the purpose of flavor improvement, enzyme inactivation and pasteurization product at 10$0^{\circ}C$ for 1 hour, with 4% glucose. To make improvement of rheological properties, used complex additive with 0.5% alginic acid, 1% pectin and 0.2% agar were very effective. And stability of mixing was 98.1% after centrifuged at 10,000 rpm for 60 minutes. The chemical composition of moisture, total carbohydrate, total nitrogen and amino type nitrogen in the fermented squeeze-type cockle shell by-product paste were 57.7%, 20.6%, 1,458mg% and 1,187mg%, respectively. And the ratio of amino type nitrogen in total nitrogen was 81.4%.

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The Separation of Transglutaminase Produced from Streptomyces mobaraensis and Its Application on Model Food System (Streptomyces mobaraensis로부터 생산되는 transglutaminase 분리 및 모델식품 적용)

  • Yoo, Jae-Soo;Shin, Weon-Sun;Chun, Gie-Tack;Kim, Young-Soo;Jeong, Yong-Seob
    • Korean Journal of Food Science and Technology
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    • v.35 no.2
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    • pp.260-265
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    • 2003
  • Transglutaminase (TG) was prepared from Streptomyces mobaraensis to improve texture and self-life of food. In preliminary experiments, texture of the dough was not improved due to the interference in microbial TG reaction by proteases present in the crude enzyme. Among the cation exchange resins tested for the removal of proteases, MonoPlus S 100 was the most efficient. Further purification steps with a quaternary ammonia salt resin and gel permeation chromatography effectively removed proteases from crude enzyme. Molecular weight of purified enzyme was about 38,000 on SDS-polyacrylamide gel electrophoresis. Farinograph data showed the addition of purified enzyme to wheat flour gave higher stability and lower weakness values those that of crude enzyme.

Characterization of the Bacteriocin from Enterococcus faecium CJNU 2008 (Enterococcus faecium CJNU 2008 균주 생산 박테리오신의 특성 규명)

  • Seo, Souk-Jin;Yang, Jung-Mo;Moon, Gi-Seong
    • Journal of Food Hygiene and Safety
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    • v.33 no.6
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    • pp.516-520
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    • 2018
  • Bacteriocin is a proteinaceous compound produced by microorganisms showing antimicrobial activities. In this study, the physicochemical properties of the bacteriocin produced by Enterococcus faecium CJNU 2008 strain were characterized. Partially purified bacteriocin showed stabilities against heat treatments at $100^{\circ}C$ for 30 min and $121^{\circ}C$ for 15 min and against solvents treatments such as methanol, ethanol, acetone, acetonitrile and chloroform. The bacteriocin also exhibited stabilities against lipase and ${\alpha}-amylase$ treatments but the stability was abolished at protease treatment, indicating that the antimicrobial agent from E. faecium CJNU 2008 was a proteinaceous bacteriocin. The bacteriocin also showed bactericidal mode of action against Listeria monocytogenes. The molecular mass of the bacteriocin was estimated to be under 6.5 kDa by a tricine-SDS-PAGE analysis. The bacteriocin was purified by HPLC. Further studies toward biochemical analysis of the bacteriocin are needed in near future.

Hydrolysis of Egg Yolk Protein in a Packed Bed Reactor by Immobilized Enzyme (충진층 반응기에서 고정화 효소에 의한 난황 단백질의 가수분해)

  • Kang, Byung-Chul
    • Journal of Life Science
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    • v.20 no.11
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    • pp.1656-1661
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    • 2010
  • Alkaline protease for the hydrolysis of egg yolk protein was immobilized on five carriers - Duolite A568, Celite R640, Dowex-1, Dowex 50W and Silica gel R60. Duolite A568 showed a maximum immobilization yield of 24.7%. Optimum pH for the free and immobilized enzyme was pH 8 and 9, respectively. However, no change was observed in optimum temperature ($50^{\circ}C$). Thermal stability was observed in immobilized enzymes compared to free enzymes. The immobilized enzyme retained 86% activity after 10 cycle operations in a repeated batch process. The effect of flow rate on the stability of enzyme activity in continuous packed-bed reactor was investigated. Lowering flow rate increased the stability of the immobilized enzyme. After 96 hr of continuous operation in a packed-bed reactor, the immobilized enzyme retained 83 and 61% activity when casein and egg yolk were used as a raw materials, respectively.

Effective Components of Commercial Enzyme Food Products and Their HACCP Scheme (시판 효소식품의 유용성분과 HACCP 관리 방안에 관한 연구)

  • Lee, Eun-Joo;Lee, Cherl-Ho
    • Korean Journal of Food Science and Technology
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    • v.33 no.4
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    • pp.461-468
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    • 2001
  • The effectiveness and safety of Enzyme Food, a group of dietary supplements designated by Korean Food Law, were evaluated and the possibility of HACCP (Hazard Analysis Critical Control Point) application was investigated. Chemical composition, enzyme activities and the degree of bacterial contamination in 12 samples of different brands sold in Korean market were measured. The chemical composition of the selected products varied and inconsistent to those claimed in the label description. It is known that effectiveness of Enzyme Food depends on enzyme activity, but enzyme activities of ${\alpha}-amylase$ varied from $1,793\;{\mu}g/min$ g to $159\;{\mu}g/min$ g and those of ${\beta}-amylase$ ranged from $171\;{\mu}g/min$ g to $11\;{\mu}g/min$ g. The protease activities varied from $27.57\;{\mu}g/min$ g to $0.18\;{\mu}g/min$ g. In coli-form bacterial test, positive reactions were appeared in the 50 % of the samples. Numbers of bacteria ranged from $1.3\;{\times}\;10^5\;to\;1.2\;{\times}\;10^9$. Five CCPs were identified; heating, inoculation, cultivation, drying and granulation. Consideration of HACCP system indicated that the pretreatment of raw material, checking of bacterial contamination and stability of enzyme activity during fermentation process were important factors for the quality of Enzyme Foods.

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Preparation of Branched-chain Amino Acid (BCAA)-enriched Hydrolysates from Corn Gluten (고 분지아미노산 함유한 옥수수 단백가수물의 제조조건 탐색)

  • Chung, Yong-Il;Bae, In-Young;Lee, Hyeon-Gyu
    • Korean Journal of Food Science and Technology
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    • v.42 no.1
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    • pp.39-44
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    • 2010
  • The process of the preparation of branched-chain amino acid (BCAA)-enriched hydrolysates from corn gluten was optimized through the parameters of pre-treatment (heating and cellulosic hydrolysis), hydrolysis method (acid, protease, and microbe plus protease), concentration, and spray drying condition. The protein yield of corn gluten was increased by heating and cellulase treatments. Among three different hydrolysis methods, the combined use of microbes and protease was the most effective in terms of free amino acid (FAA) and BCAA content of the corn gluten hydrolysates. In addition, the FAA and BCAA content in the hydrolysates prepared by microbial and enzymatic combined treatment were improved by a concentration process. Spray drying conditions for the preparation of the powder from the hydrolyzed reactant were an inlet temperature of $185^{\circ}C$, outlet temperature of $80^{\circ}C$, and the use of maltodextrin as an anticaking agent. Thus, this study established an economical process for preparation of value-added hydrolysates of excellent productivity and quality, in terms of high BCAA content and product stability.

Changes of Chemical Properties in Processing of Low Salted and Fermented Shrimp Using Gamma Irradiation immediately before Optimum Fermentation (저염 새우젓 제조를 위한 최적 숙성직전의 감마선 조사시 화학적 품질변화)

  • Lee, Kyong-Haeng;Ahn, Hyun-Joo;Lee, Cherl-Ho;Kim, Yeung-Ji;Byun, Myung-Woo
    • Korean Journal of Food Science and Technology
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    • v.32 no.5
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    • pp.1051-1057
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    • 2000
  • Gamma irradiation was applied to develop fermented shrimp product with lower salt concentration, high sensory quality and storage stability. Shrimp was prepared with 15 and 20% of salt and fermented at $15^{\circ}C$. The sample was irradiated for 15% added salt at the 4th week and for 20% at the 6th week during fermentation with 0, 5 and 10 kGy absorbed doses. The irradiation was applied at optimum stage of fermentation determined when the content of amino nitrogen(AN) arrived approximately 400 mg%. Chemical properties such as amino nitrogen(AN), volatile base nitrogen(VBN), trimethylamine(TMA) and neutral protease activity were examined during whole fermentation. The AN, VBN, TMA and protease activity were not affected immediately after gamma irradiation. The more NaCl concentrations and irradiation dose, the less content of chemical compounds and protease activity was found. From the results of chemical properties, it was concluded that fermented shrimp with 15% of salt and irradiated at 10 kGy before optimum fermentation, or 20% and 5 kGy or above were maintained the sound quality during storage compared with the control.

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Incubation Conditions and Physico-Chemical Factors Affecting Aflatoxin B1 Binding of Lactic Acid Bacteria (Aflatoxin B1에 대한 유산균의 결합력에 영향을 미치는 배양조건과 물리화학적 인자)

  • Lim, Sung-Mee;Ahn, Dong-Hyun
    • Korean Journal of Microbiology
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    • v.49 no.3
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    • pp.253-261
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    • 2013
  • The purpose of this study was to investigate the aflatoxin $B_1$ binding of lactic acid bacteria (LAB) isolated from Korean traditional soybean paste and to evaluate the effect of incubation conditions and physico-chemical factors on the binding ability of LAB to this mutagen. The amount of aflatoxin $B_1$ bound by Enterococcus faecium DJ22, Lactobacillus fermentum DJ35, Lactobacillus rhamnosus DJ42, and Lactobacillus pentosus DJ47 was strain specific with the percent bound ranging from 19.3% to 52.1%. However, Enterococcus faecalis DJ14, Lactobacillus panis DJ29, and Pediococcus halophilus DJ50 strains did not exhibit any of the binding ability to aflatoxin $B_1$. For most strains, the binding ability was significantly affected by the environmental conditions such as the aflatoxin $B_1$ level, incubation time and temperature, and the initial cell count of LAB. The stability of the aflatoxin $B_1$-bacteria complexes was significantly more unstable after washing. In addition, the binding stability between viable and nonviable cells was not statistically significant. Treatment with heating, acidic pH, ${\alpha}$-amylase, protease, lysozyme, or sodium metaperiodate caused a significant (P<0.05) decrease in aflatoxin $B_1$ binding for the tested strains, suggesting that carbohydrates or proteins in the cell walls may be involved in aflatoxin $B_1$ binding ability. Since the aflatoxin $B_1$ binding of LAB was significantly reduced (P<0.05) by the pretreatment of the urea, the binding force observed in this study may have resulted from hydrophobic interaction.

Effects of Pretense Treatment on Functional Properties of Soymilk Protein (단백분해 효소처리가 두유단백질의 기능성에 미치는 영향)

  • 변진원;황인경
    • Korean journal of food and cookery science
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    • v.11 no.1
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    • pp.26-32
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    • 1995
  • This study was carried out to investigate the effect of protease on the functionality of soymilk protein. The protease from Bacillus polymyxa was selected because of the least production of bitter taste and calcium-aggregation. The results are summarized as follows: 1. Solubility of SMP(soymilk protein) and SPI(soyprotein isolate) were lowest at pH 4.7 and increased as the pH value reached closer to either ends. PT-SMP(pretense treated soymilk protein) showed higher solubility at all pH range, especially at pH 4.7 than SMP, SPI. 2. Emulsion activity of three samples was lowest at pH 4.7 and significantly increased as pH approched higher acidic or alkaline regions. PT-SMP showed similar activity to other samples, but less stability. 3. Foam capacity of PT-SMP was lowest at pH 8 and increased in acidic, alkaline pH. PT-SMP showed higher foam capacity at all pH range, but lower foam stability than SMP and SPI. 4. PT-SMP showed higher heat coagulability than other samples at all pH range except pH 4.7.

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