• 제목/요약/키워드: enzymatic activity

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Polysaccharide Degrading Enzyme을 이용한 참모자반 효소분해 추출물의 생리활성 연구 (Biological Analysis of Enzymatic Extracts from Sargassum fulvellum Using Polysaccharide Degrading Enzyme)

  • 조은경;강수희;최영주
    • KSBB Journal
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    • 제28권6호
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    • pp.349-355
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    • 2013
  • SC092 strain, producing a polysaccharide degrading enzyme, was isolated from the seawater. This strain was identified as Microbulbifer sp. using the comparative sequence analysis against known 16S rRNA sequence. A polysaccharide degrading enzyme from this strain was used to acquire the enzymatic extracts of Sargassum fulvellum. DPPH radical scavenging and SOD activity of the enzyme extracts of S. fulvellum were about 61.9% and 82.9% at 2 mg/mL, respectively. Nitrite scavenging activities was 52.5% at 2 mg/mL on pH 1.2. In addition, ${\alpha}$-glucosidase inhibitory activity was also increased in a dose-dependent manner and was about 52.7% at 2 mg/mL. To determine the influence of enzyme extracts of S. fulvellum on alcohol metabolism, the generating activity of reduced-nicotinamide adenine dinucleotide (NADH) by alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) were measured. ADH and ALDH activities were 118.0% and 177% at 2 mg/mL, respectively. ${\alpha}$-glucosidase inhibitory activity of enzyme extracts of S. fulvellum was remarkably increased in a dose-dependent manner and was about 52.7% at 2 mg/mL. These results indicate alcoholizing and ${\alpha}$-glucosidase inhibitory activities can be enhanced by the enzymatic extracts of S. fulvellum.

Correlation between pr1 and pr2 Gene Content and Virulence in Metarhizium anisopliae Strains

  • Rosas-Garcia, Ninfa M.;Avalos-de-Leon, Osvaldo;Villegas-Mendoza, Jesus M.;Mireles-Martinez, Maribel;Barboza-Corona, J.E.;Castaneda-Ramirez, J.C.
    • Journal of Microbiology and Biotechnology
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    • 제24권11호
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    • pp.1495-1502
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    • 2014
  • Metarhizium anisopliae is a widely studied model to understand the virulence factors that participate in pathogenicity. Proteases such as subtilisin-like enzymes (Pr1) and trypsin-like enzymes (Pr2) are considered important factors for insect cuticle degradation. In four M. anisopliae strains (798, 6342, 6345, and 6347), the presence of pr1 and pr2 genes, as well as the enzymatic activity of these genes, was correlated with their virulence against two different insect pests. The 11 pr1 genes (A, B, C, D, E, F, G, H, I, J, and K) and pr2 gene were found in all strains. The activity of individual Pr1 and Pr2 proteases exhibited variation in time (24, 48, 72, and 96 h) and in the presence or absence of chitin as the inductor. The highest Pr1 enzymatic activity was shown by strain 798 at 48 h with chitin. The highest Pr2 enzymatic activity was exhibited by the 6342 and 6347 strains, both grown with chitin at 24 and 48 h, respectively. Highest mortality on S. exigua was caused by strain 6342 at 48 h, and strains 6342, 6345, and 6347 caused the highest mortality 7 days later. Mortality on Prosapia reached 30% without variation. The presence of subtilisin and trypsin genes and the activity of these proteases in M. anisopliae strains cannot be associated with the virulence against the two insect pests. Probably, subtilisin and trypsin enzyme production is not a vital factor for pathogenicity, but its contribution is important to the pathogenicity process.

Characterization of Bone Marrow Cell Proliferating Arabinogalactan through Peyer`s Patch Cells from Rhizomes of Atractylodes lancea DC

  • Yu, Kwang-Won;Hwang, Jong-Hyun
    • Preventive Nutrition and Food Science
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    • 제6권3호
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    • pp.180-186
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    • 2001
  • Bone marrow cell proliferating arabinogalactan-like polysaccharide (ALR-3IIa-1-1) has been purified from rhizomes of Atractylodes lancea DC. In order to characterize the essential structure of ALR-3IIa-1-1 for expression of the activity, sequential enzymatic digestion using ego-$\alpha$-L-arabinofurasidase (AFase) and ego-$\beta$-D-(1longrightarrow3)-galactanase (GNase) was employed. After ALR-3IIa-1-1 was digested with the AFase, the GNase digestion cleaved only 10% and 23% of 3-linked and 3,6-branched galactose, respectively, from arabinose-trimmed ALR-3IIa-1-1 (AT-ALR-3IIa-1-1), and gave small amounts of intermediate size (AT-G-2) and shorter oligosaccharides (AT-G-3) fractions in addition to a large amount of the GNase resistant fraction (AT-G-1). When AT-G-1 was redigested gradually with the AFase and GNase, it released trace amounts of oligosaccharides in addition to a large amount of the resistant fraction. When the final enzyme-resistant fraction from AT-G-1 was digested simultaneously with both AFase and GNase, the resistant fraction was significantly degraded into two long fragments (3AT-3G-1 and 2). The mixture of digestion products from the first GNase digestion of AT-ALR-3IIa-1-1 showed a significantly decreased bone marrow cell proliferation activity to about 30% of the activity of ALR-3IIa-1-1, but the GNase resistant fraction (AT-7-1) still had significant activity. Although the second gradual enzymatic digestion of AT-G-1 showed a marginal decrease in activity, the resulting fragments (3AT-3G-1 and 2) by the final simultaneous enzymatic digestion lost most of the activity. Component sugar, methylation and FAB-MS analyses indicated that the digestion products (AT-G-21 AT-G-31 2AT-2G-2 and 2AT-2G-3) released from AT-ALR-3IIa-1-1 by the sequential enzymatic digestion contained galactose-containing oligosaccharides mainly comprising 6-linked galactose, that some of which were partially arabinosylated, and these oligosaccharides were attached to $\beta$-D-(1longrightarrow3)-galactan backbone in its non-reducing terminal side as side chains.

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재래종 황색자두효소 갈변반응 생성물의 돌연변이 억제작용 (Desmutagenicity of the Enzymatic Browning Reaction Products Which Obtained from Prunus salicina (yellow) Enzyme and Polyphenol Compounds)

  • 함승시
    • Applied Biological Chemistry
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    • 제30권1호
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    • pp.71-76
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    • 1987
  • 재래종 황색계자두 효소와 4종류의 polyphenol 화합물과 반응시켜 얻어진 효소적 갈변반응 생성물에 대하여 변이원성 및 변이원 억제작용에 관한 실험결과 B. subtilis H17과 M45 두 균주를 이용한 rec-assay에서 4종류의 갈변반응 생성물 모두 변이원성은 없었다. rec-assay에서 금속이온의 영향을 실험한 결과 pyrogallol의 갈변반응 생성물의 경우 $Fe^{3+},\;Mn^{2+},\;Zn^{2+},\;Ni^{2+}\;Al^{2+}$에서 약한 영향을 받았으며 hydroxyhydroquinone의 갈변반응 생성물에서는 $Cu^{2+},\;Pb^{2+}$, 그리고 catechol의 갈변반응 생성물에서는 $Mn^{2+}$ 공존하에서 약한 영향을 받았다. DNA절단실험 결과 갈변반응 생성물 자체에 의해서는 4종류의 시료 모두 DNA절단능력은 없었다. 금속이온의 영향에서는 $Fe^{2+}$의 경우 그 자체에 의해 절단되었으며 $Cu^{2+}$은 DNA 절단을 촉진하는 것으로 나타났다. S-9 mix를 첨가한 돌연변이 유발실험에서는 4종류의 갈변반응 생성물 모두 농도증가에도 변이원성은 없었으며 변이원 물질인 $benzo\;{\alpha}\;pyrene$에 대한 돌연변이원 억제반응에서는 4종류의 갈변반응 생성물 모두 농도증가에 따라 대조구에 비해 강한 억제작용을 나타냈다. 따라서 재래종 황색계 자두로부터 추출한 polyphenol oxidase와 4종류의 polyphenol 화합물과 반응시켜 얻어진 효소적 갈변반응 생성물의 들연변이원성 실험결과 갈변반응 생성물자체에 의한 돌연변이원성은 없었으며 금속이온의 영향도 받지 않는 것으로 나타났으나 강력한 발암물질인 $benzo\;{\alpha}\;pyrene$을 사용한 돌연변이 억제실험에서는 4종류의 갈변반응 생성물 모두 강한 돌연변이 억제작용이 있음을 알 수 있었다.

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카페인산의 효소적 산화반응으로부터 췌장 지방분해효소 저해 물질의 분리 (Secondary Metabolites from Enzymatic Oxidation of Caffeic Acid with Pancreatic Lipase Inhibitory Activity)

  • 김태훈;김명권
    • 한국식품영양과학회지
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    • 제44권12호
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    • pp.1912-1917
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    • 2015
  • 천연식물에 광범위하게 존재하는 대표적인 페닐프로파노이드 화합물인 caffeic acid에 대해 배 유래의 polyphenol oxidase로 산화반응을 수행하여 상대적으로 높은 pancreatic lipase 저해 활성($IC_{50}$; $161.2{\pm}2.8{\mu}g/mL$)을 확인하였으며, 이는 caffeic acid와 비교하였을 경우 활성이 상승함을 알 수 있었다. Caffeic acid 산화반응물에 대해서 $C_{18}$ 겔을 활용한 column chromatography를 수행하여 4종의 리그난 화합물을 분리하였고, 각 화합물의 화학구조는 NMR 스펙트럼 데이터 해석 및 표품과의 HPLC 직접 비교를 통하여 phellinsin A(2), caffeicinic acid(3), isocaffeicinic acid(4), 7,8-erythro-caffeicin(5)으로 동정하였다. 이들 화합물중 phellinsin A(2)는 $IC_{50}$ 값이 $66.3{\pm}2.6{\mu}M$로 가장 강한 효능을 나타내었으며, 다음으로 caffeic acid 2분자의 산화 결합을 통해 생합성된 caffeicinic acid(3)의 $IC_{50}$ 값이 $109.6{\pm}3.7{\mu}M$의 저해능을 나타내었다. 배에 존재하는 polyphenol 산화효소에 의해 생합성된 caffeic acid 이량체가 pancreatic lipase 저해 활성 물질임을 확인하였으며, 이들 활성은 caffeic acid가 결합 양상에 따른 화합물의 구조에 따라 다름이 시사되었다. 향후 이들 활성물질의 활성 기작에 대한 연구가 필요하며 본 연구 결과는 보다 우수한 pancreatic lipase 저해능을 가지는 새로운 선도화합물 발굴을 위한 기초자료로 이용될 수 있을 뿐만 아니라 항비만 물질의 상업화를 위한 기초자료로 이용될 수 있을 것으로 사료된다.

Purification and Characterization of Soybean Cotyledonary Spermidine Dehydrogenase

  • Park, Sung-Joon;Cho, Young-Dong
    • BMB Reports
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    • 제28권5호
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    • pp.408-413
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    • 1995
  • Decrease in the amount of cotyledonary spermidine in Glycine max under anaerobic conditions related to an increase in spermidine dehydrogenase. Under the same conditions, no enzymatic activity of diamine oxidase was observed. Exposure of Glycine max both to spermidine and 1,3-diaminopropane under anaerobic conditions resulted in a decrease in spermidine contents. Correlated with the decrease in spermidine contents, there was a drastic increase in spermidine dehydrogenase. The molecular weight of the purified enzyme estimated by Sephacryl S-300 gel column and SDS gel electrophoresis were 130,000 dalton and 65,000 dalton, respectively, indicating that the enzyme is a dimer. The optimal pH for activity was 9.3. The $K_m$ value for spermidine was 0.61 mM. Neither metal ions nor polyamine and derivatives affected enzymatic activity, but the enzyme was inhibited by DTNB, NEM and PCMB, suggesting that a cysteine residue of the enzyme is associated with or involved in enzyme activity. To our knowledge, this is the first report describing properties of the enzyme from plants. Considered together, the data in this paper indicate that both spermidine and 1,3-diaminopropane, novel activators, enhance the spermidine dehydrogenase activity and control the intracellular spermidine contents.

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오미자 효소가수분해물에 의한 장내 세균총 개선 및 장관 염증 억제 효과의 효소학적 평가 (Enzymological Evaluation of Bowel Inflammation Inhibitory Activity and Intestinal Microbial Flora Improvement by Enzymatic Hydrolysate of Schizandrae Fructus)

  • 류일환;권태오;이강수;윤용갑
    • 생약학회지
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    • 제38권4호
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    • pp.363-371
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    • 2007
  • Schizandrae Fruits has been used as a traditional Oriental medicine for treatment of many stress-induced diseases. In the present study, we investigated inhibitory activity of enzymatic hydrolysate of Schizandrae fructus (SC-EX) in growth of tested intestinal microorganism and activity of bowel inflammation related enzyme. SC-EX was added to the proteose peptone-yeast extract-fildes (PYF) media to investigation the effect on the growth of type culture of intestinal microorganism. The growth of lactic acid bacteria such as Bifidobacterium species and Lactobacillus species was accelerated by more than 3% concentration of SC-EX. But, growth of harmfulness bacteria such as E.coli, Clostridium sp. Staphylococcus sp. Streptococcus sp. was inhibited by more than 3% concentration of SC-EX. Also, SC-EX was exhibited a concentration-dependent inhibitory activity of the bowel inflammation related enzymes. The SC-EX was showed 76% and 92% inhibitory activity of 5-lipoxygenase and cyclooygenase at 5% additional concentration respectively. Our results indicated that SC-EX may possess improvement effect on the intestinal flora and Anti-inflammatory effect on the bowel.

Effect of Tween 80 on Hydrolytic Activity and Substrate Accessibility of Carbohydrolase I (CBH I) from Trichoderma viride

  • Kim, Wanjae;Gamo, Yuko;Sani, Yahaya Mohammed;Wusiman, Yimiti;Ogawa, Satoru;Karita, Shuichi;Goto, Masakazu
    • Asian-Australasian Journal of Animal Sciences
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    • 제19권5호
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    • pp.684-689
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    • 2006
  • The present study examined the effects of Tween 80 on the attachment and hydrolytic activity of a cellulase enzyme against ball-milled cellulose (BMC), using the whole component (native CBH I) and the catalysis module (core CBH I) of carbohydrolase I purified from Trichoderma viride (Meicelase, Meiji Seika, Tokyo, Japan). The effects were evaluated as protein concentrations in the supernatant after mixing enzyme and substrate with Tween 80 at room temperature. Tween 80 decreased the adsorption of native CBH I and core CBH I onto BMC (p<0.001) and increased the amount of reducing sugars released from BMC by native CBH I (p<0.001). However, Tween 80 did not enhance the hydrolytic activity of core CBH I. Observations using SEM revealed that Tween 80 caused cellulose filter paper to swell and enhanced surface cracks and filaments caused by native CBH I but not by core CBH I. These results suggested that Tween 80 decreases enzyme adsorption to its substrate but enhances enzymatic activity.

Computational identification of significantly regulated metabolic reactions by integration of data on enzyme activity and gene expression

  • Nam, Ho-Jung;Ryu, Tae-Woo;Lee, Ki-Young;Kim, Sang-Woo;Lee, Do-Heon
    • BMB Reports
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    • 제41권8호
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    • pp.609-614
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    • 2008
  • The concentrations and catalytic activities of enzymes control metabolic rates. Previous studies have focused on enzyme concentrations because there are no genome-wide techniques used for the measurement of enzyme activity. We propose a method for evaluating the significance of enzyme activity by integrating metabolic network topologies and genome-wide microarray gene expression profiles. We quantified the enzymatic activity of reactions and report the 388 significant reactions in five perturbation datasets. For the 388 enzymatic reactions, we identified 70 that were significantly regulated (P-value < 0.001). Thirty-one of these reactions were part of anaerobic metabolism, 23 were part of low-pH aerobic metabolism, 8 were part of high-pH anaerobic metabolism, 3 were part of low-pH aerobic reactions, and 5 were part of high-pH anaerobic metabolism.

Effects of Chitosan and Lactic Acid on Enzymatic Activities and Bioactive Compounds during Germination of Black Rice

  • Kim, Kwan-Soo;Jang, Hae-Dong
    • Preventive Nutrition and Food Science
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    • 제9권3호
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    • pp.199-205
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    • 2004
  • The effect of chitosan on enzymatic activities and on bioactive compounds was characterized during germination at $25^{\circ}C$ for 7 days to search for a method to produce a germinated black rice. The germination rate was reduced by the addition of lactate and chitosan. The rotting rate was greatly decreased by chitosan, suggesting that the addition of chitosan into a germination solution might be an effective method for controlling fungal contamination during the germination of cereals. The addition of 100 and 200 ppm chitosan increased $\alpha$-amylase activity after 7 days by up to 152 % and 197 %, respectively. The activities of $\beta$-amylase and $\beta$-glucosidase were lower with 200 ppm chitosan than in distilled water and 100 ppm lactate. The amount of total soluble phenolics and total flavonoids decreased rapidly for four days and thereafter remained constant until the seventh day. The antioxidant activity of germinated black rice, in terms of hydrogen-donating activity, increased slowly and did not correspond to the changes of total soluble phenolics and total flavonoids. The amount of phytic acid was reduced by the addition of 200 ppm chitosan compared to distilled water, indicating that chitosan could be used as an elicitor for the increase of phytase activity during the germination of black rice.