• 제목/요약/키워드: enzyme hydrolysis

검색결과 969건 처리시간 0.024초

Cellulase 성분 중 Endo-gluanasec의 반응 및 흡착특성에 관한 연구 (Reaction Kinetics and Absorption Property of Low Molecular Weight Endo-glucanase Component of Cellulase)

  • 류왕식;유두영
    • 한국미생물·생명공학회지
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    • 제8권1호
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    • pp.41-46
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    • 1980
  • 저분자량의 효소인 셀루라아제가 Sephadex G-100 gel chromatography를 사용하여 정제되었다. 정제된 성분의 생화학적 성질들이 조사되었는데 최적 PH와 온도가 각각 6.0과 5$0^{\circ}C$이었다. 서로 다른 결정도(crystallinity)를 갖고 있는 4가지 섬유소 기질의, 효소에 의한 가수분해가 측정되었다. 무정형(amorphous) 부분의 가수분해가 일어난 후에 결정화되어 있는 부분의 가수분해가 뒤따라온다는 것을 알 수 있었다. 효소가 섬유소 기질에 흡착되는 정도가 가수분해 반응에 미치는 영향을 보기 위해 흡착에 대한 연구가 수행되었다. 시간에 따라서 소분자량의 endo-glucanase가 여러가지섬유소 기질에 흡착되는 정도의 변화가 25분간 측정되었다. 무정형의 섬유소에 흡착되는 속도와 정도가 결정형 섬유소에 대한 그것들 보다 더욱 크게 나타났다. 이러한 결과는 endo-glucanase가 섬유소의 결정화 부분의 가수분해보다는 무정형부분의 가수분해에 대해서 더욱 중요한 역할을 한다는 것을 시사해 준다.

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Affinity Labeling of E. coli GTP Cyclohydrolase I by a Dialdehyde Derivative of Guanosine Triphosphate

  • Ahn, Chi-Young;Park, Sang-Ick;Kim, Ju-Myeong;Yim, Jeong-Bin
    • BMB Reports
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    • 제28권1호
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    • pp.72-78
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    • 1995
  • Time-dependent inactivation of E. coli GTP cyclohydrolase I with a 2',3'-dialdehyde derivative of GTP (oGTP) was directed to the active site of the enzyme, and was dependent on the concentration of oGTP. The kinetics of inactivation were biphasic with a rapid reaction occurring immediately upon exposure of the enzyme to oGTP followed by a slow rate of inactivation. The $K_i$ value of oGTP for the enzyme was 0.25 mM. Inactivation was prevented by preincubation of the enzyme with GTP, the substrate of the enzyme. At 100% inactivation, 2.3 mol of [8.5'-$^3H$]oGTP were bound per each enzyme subunit, which consists of two identical polypeptides. The active site residue which reacted with the affinity label was lysine. oGTP interacted selectively with the ${\varepsilon}$-amino group of lysine in the GTP-binding site to form a morpholine-like structure which was stable without sodium borohydride treatment. However, triphosphate group was eliminated during the hydrolysis step. To identify the active site of the enzyme, [8.5'-$^3H$]oGTP-labeled enzyme was cleaved by endoproteinase Lys-C, and the $^3H$-labeled peptide was purified by HPLC. The amino acid sequence of the active site peptide was Pro-Ser-Leu-Ser-Lys, which corresponds to the aminoterminal sequence of the enzyme.

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Kinetic Modeling of Simultaneous Saccharification and Fermentation for Ethanol Production Using Steam-Exploded Wood with Glucose- and Cellobiose-Fermenting Yease, Brettanomyces custersii

  • Moon, Hyun-Soo;Kim, Jun-Seok;Oh, Kyeong-Keun;Kim, Seung-Wook;Hong, Suk-In
    • Journal of Microbiology and Biotechnology
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    • 제11권4호
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    • pp.598-606
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    • 2001
  • A mathematical model is proposed that can depict the kinetics of simultaneous saccharification and fermentation (SSF) using steam-exploded wood(SEW) with a glucose- and cellobiose-fermenting yeast strain. Brettanomyces custersii. An expression to describe the reduction of the relative digestibility during the hydrolysis of the SEW is introduced in the hydrolysis model. The fermentation model also takes two new factors into account, that is, the effects of the inhibitory compounds present in the SEW hydrolysates on the microorganism and the fermenting ability of Brettanomyces custersii, which can use both glucose and cellobiose as carbon sources. The model equations were used to simulate the hydrolysis of the SEW, the fermentation of the SEW hydrolysates, and a batch SSF, and the results were compared with the experimental data. The model was found to be capable of representing ethanol production over a range of substrate concentrations. Accordingly, the limiting factors in ethanol production by SSF under the high concentration of the SEW were identified as the effect of inhibitory compounds present in the SEW, the enzyme deactivation, and a limitation in the digestibility based on the physical condition of the substrate.

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Antioxidative Activities of Hydrolysates from Duck Egg White Using Enzymatic Hydrolysis

  • Chen, Yi-Chao;Chang, Hsi-Shan;Wang, Cheng-Taung;Cheng, Fu-Yuan
    • Asian-Australasian Journal of Animal Sciences
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    • 제22권11호
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    • pp.1587-1593
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    • 2009
  • Duck egg white (DEW) hydrolysates were prepared by five enzymes (papain, trypsin, chymotrypsin, alcalase, and flavourzyme) and their antioxidant activities investigated in this study. DEW hydrolyzed with papain (DEWHP) had the highest peptide content among the five enzymatic treatments. Besides, the peptide content of DEWHP increased when the enzyme to substrate ratio (E/S ratio) increased. It was suggested that higher E/S ratio contributed to elevate the degree of hydrolysis of DEW effectively. Similar results were also obtained by Tricine-SDS-PAGE. In addition, SDS-PAGE patterns indicated papain was the only one amongst all enzymes with the ability to hydrolyze DEW. In antioxidant properties, DEWHP showed more than 70% of inhibitory activity on linoleic acid peroxidation and superoxide anion scavenging. Moreover, the $Fe^{2+}$ chelating effect of DEWHP was greater than 90%, while no significant difference was observed in DPPH radical scavenging and reducing ability. The results of peptide contents, antioxidant activities and electrophoresis suggested that the higher the peptide content, the stronger the antioxidant activities in DEWHP.

Physicochemical Properties of Gamma-Irradiated Corn Starch

  • Lee, Yong-Jin;Kim, Sun-Young;Lim, Seung-Taik;Han, Sag-Myung;Kim, Hye-Mi;Kang, Il-Jun
    • Preventive Nutrition and Food Science
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    • 제11권2호
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    • pp.146-154
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    • 2006
  • Structural modification of corn starch by gamma irradiation was evaluated for under dry conditions at varied intensities from 0 to 40 kGy. Under scanning electron microscopy, the granule shape of corn starch was not significantly affected by the irradiation up to 40 kGy. In addition, X-ray diffraction and melting patterns of the irradiated starches were similar to those of the native starch, indicating that crystalline regions in the starch granules were not changed by irradiation. However, the pattern of gel permeation column chromatography showed a significant increase in partial hydrolysis of gamma irradiated starch samples. The degree of polymerization and the paste viscosity of irradiated starch samples dose-dependently decreased significantly with irradiation, and increased solubility and clarity were observed in the irradiated starch solution. In addition, the degree of retrogradation decreased as irradiation dose increased. Irradiation of corn starch has advantages over the ordinary acid or the enzyme hydrolysis modification methods. It does not affect the granular shape and crystalline phase of starch during hydrolysis, and the process can be carried out in dry state.

차 부산물로부터 효소를 이용한 캠페롤 생산 (Production of kaempferol by enzymatic hydrolysis of tea seed extract)

  • 임윤영;김은기
    • KSBB Journal
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    • 제23권2호
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    • pp.131-134
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    • 2008
  • 캠페롤은 항산화, 항염등의 성질을 가지고 있지만 생산이 쉽지가 않다. 차의 씨를 가공하고 발생하는 부산물 (tea seed extract)에는 캠페롤과 당이 결합되어 있는 형태인 camelliaside A와 camelliaside B가 포함되어 있다. 본 실험에서는 캠페롤을 효소의 가수분해에 의하여 생산하였다. 가장 효과적인 효소는 $pectinex^{(R)}100L$ 이며, 효소 반응에서 캠페롤은 48시간내에 생산되었다. 반응 최적 조건은 $40^{\circ}C$, pH 4이며 생산의 최적조건하에서 차 부산물 1 kg당 1.6 g의 캠페롤이 생산되었고 약 80%의 캠페롤이 침전되었다. 효소반응에 의한 캠페롤 생산법은 생산물의 회수가 용이한 효과적인 방법임을 알 수 있다.

해바라기씨 단백질에서 plastein의 합성 (Plastein formation from sunflower seed protein)

  • 노재문;김재욱
    • Applied Biological Chemistry
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    • 제34권1호
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    • pp.1-7
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    • 1991
  • 해바라기씨 단백질에서 plastein을 합성 이용하기 위해 먼저 해바라기씨를 pepsin을 이용해 가수분해하는 최적 조건과, plastein을 합성하는 최적 조건을 구하였다. 최적 가수분해 조건은 pH 1.5, $45^{\circ}C$, 2% 기질농도와 2% pepsin 농도에서 24시간 반응시키는 것이었고 plastein 합성의 최적조건은 기질농도 50%, pH 4.5, $50^{\circ}C$, 0.25% pepsin 농도에서 18시간 반응시키는 것이었다. Plastein의 생성을 확인하기 위해 thin layer chromatography를 실시한 결과 해바라기씨 농축가수분해물의 TLC pattern과 plastein의 TLC pattern이 하나의 spot를 제외하고는 다르게 나타났는데 이는 plastein과 기질이 다른 것임을 표시하는 것이었다.

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Effect of Enzymatic Hydrolysis of 7S Globulin, a Soybean Protein, on Its Allergenicity and Identification of its Allergenic Hydrolyzed Fragments Using SDS-PAGE

  • Keum, Eun-Hee;Lee, Sang-Il;Oh, Sang-Suk
    • Food Science and Biotechnology
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    • 제15권1호
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    • pp.128-132
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    • 2006
  • This study was undertaken to investigate the effect of peptic and chymotryptic hydrolyses of 7S globulin, the major allergen of soybean protein, on its allergenicity, as measured by enzyme linked immunosorbent assay (ELISA), and to identify the allergenic hydrolyzed fragments of 7S globulin using SDS-PAGE. When 7S globulin was hydrolyzed by pepsin, the allergenicity was reduced by over 50%. However, the allergenicity of 7S globulin reduced by peptic hydrolysis was recovered in the sera from 5 out of 10 patients following sequential chymotryptic hydrolysis. Two fragments, with molecular weights 20-25 and 13-16 kDa, among the hydrolysate of 7S globulin by sequential pepsin and chymotrypsin showed reactivity with sera from 10 soybean-allergenic patients. As a result of the theoretical hydrolyses of ${\beta}$-conglycinin, which is a major protein of 7S globulin, it is suggested that the 20-25 kDa fragments were the fragments of the ${\alpha}$-subunit of ${\beta}$'-conglycinin and that the 10-16 kDa fragments were from the ${\alpha}$'-subunit.

Optimization of Enzymatic Pretreatment for the Production of Fermented Ginseng using Leaves, Stems and Roots of Ginseng

  • Cho, Kyung-Lae;Woo, Hye-Jin;Lee, In-Sook;Lee, Jun-Won;Cho, Young-Cheol;Lee, Il-Nam;Chae, Hee-Jeong
    • Journal of Ginseng Research
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    • 제34권1호
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    • pp.68-75
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    • 2010
  • This study sought to optimize the extraction and enzymatic treatment conditions of Panax ginseng leaves, stems, and roots for the production of fermented ginseng. The optimization enhanced the extraction of total saccharide, a nutrient and growth-activating factor for Lactobacillus bacteria. The hydrolysis of ginseng leaves, stems, and roots was tested with eight enzymes (Pentopan, Promozyme, Celluclast, Ultraflo, Pectinex, Ceremix, Viscozyme, and Tunicase). The enzymatic hydrolysis conditions were statistically optimized by the experimental design. Optimal particle size of ginseng raw material was <0.15 mm, and optimal hydrolysis occurred at a pH of 5.0-5.5, a reaction temperature of 55-$60^{\circ}C$, a Ceremix concentration of 1%, and a reaction time of 2 hr. Ceremix produced the highest dry matter yield and total saccharide extraction. Ginseng leaves were found to be the most suitable raw material for the production of fermented ginseng because they have higher carbohydrate and crude saponin contents than ginseng roots.

유청단백질의 분리 및 단백질 분해 효소에 의한 유청단백질의 가수분해 양상 (Isolation of whey protein and hydrolysis pattern of whey protein by proteolytic enzyme)

  • 렌친핸드;배형철;정석근;남명수
    • 농업과학연구
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    • 제39권4호
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    • pp.561-568
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    • 2012
  • The aim of this study was to introduce a simple method for isolation of ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin and bovine serum albumin from cow's milk, and peptides produced by enzymatic hydrolysis of ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin and bovine serum albumin with alcalase. Whey protein were precipitated from whey by ammonium sulfate and, ${\alpha}$-lactalbumin and ${\beta}$-lactoglobulin were isolated using Hi Prep 26/60 Sephacryl S-100 column gel filtration chromatography. Bovine serum albumin and ${\beta}$-lactoglobulin were isolated by Mono-Q 5/50 GL column anion exchange chromatography of the 50% Ammonium Sulfate-supernatant. Isolated whey proteins were hydrolyzed by proteolytic alcalase. Tricine SDS-PAGE and reverse-phase HPLC analyses revealed that almost hydrolyzed all the ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin and bovine serum albumin with alcalase. Molecular weight of various peptides derived from alcalase hydrolysate were small molecular weight than 3.5 kDa.