• Title/Summary/Keyword: P-Fraction

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Studies on the Cellulase produced by Myriococcum of albomyces (Myriococcum albomyces가 생산하는 Cellulase에 관한 연구)

  • Chung, Dong-Hyo
    • Applied Biological Chemistry
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    • v.14 no.1
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    • pp.59-97
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    • 1971
  • As a study on the cellulase of Myriococcum albomyces the culture media for enzyme formation and properties of its crude preparation were investigated and the crude enzyme preparation was further fractionated. The results are summarized as follows: 1. Wheat bran solid culture produced stronger activities of cellulase than rice bran or defatted soy bean meal solid culture. 2. Shaking culture with wheat bran, rice bran or defatted soy bean meal produced higher cellulase activities than solid culture with the corresponding media. 3. The enzyme formation was higher at $45^{\circ}C$ than at $37^{\circ}C$ or $50^{\circ}C$ regardless of the kind of culture medium. 4. The formation of CMCase activity was more promoted by organic nitrogen source than inorganic nitrogen source. 5. The formation of cellulase activities were increased 1.5 to 3.0-fold by adding CMC, Avicel or cellulose powder as an inducer into 5% wheat bran basal medium. 6. Cellulase production using a tank culture procedure with addition of CMC or Avicel as an inducer was the highest at fifth day and thereafter decreased slightly. 7. The crude enzyme preparation showed pH optimum in 4.0 to 4.5, and pH stability in the range of 3.5 to 8.0. Optimum temperature for the activity was $65^{\circ}C$ which was higher than among other cellulases and it was stable at $60^{\circ}C$ for 120 minutes. 8. Dialyzed crude enzyme was activated by $Ca^{++}$ and $Mg^{++}$, but inhibited by $Hg^{++}$, $Cu^{++}$ and $Ag^{+}$. 9. Four different types of cellulase, i. e., fraction I, fraction II-a, fraction II-b, and fraction III were purified from the culture filtrate of Myriococcum albomyces through a sequence of ammonium sulfate fractionation, and elution chromatography on DEAE-Sephadex A-25, Amberlite CG-25 type 2 and hydroxyapatite columns. 10. These four cellulase fractions were showed to be homogenous by electrophoresis and ultracentrifugation and also gave a typical ultraviolet absorption spectrum of protein. 11. Four purified fraction showed different specificity toward substrates, fraction I has a stronger activity toward Avicel, cellulose powder, and gauze than that of other cellulase fractions. Fraction II-a had a powerful activity toward cellobiose but it was almost inactive agaisnt fibrous cellulose contrary to fraction I. On the contrary, the main component fraction II-b had a fairly higher activity on CMC and Avicel. Activity of fraction II-b toward cellobiose was about one-third of that of fraction II-a and activity on Avicel was lower than that of fraction I. Fraction III had a more powerful activity in decreasing viscosity of CMC. 12. Final hydrolysis products of fibrous cellulose by each fraction were cellobiose and glucose. Whereas oligosaccharides were predominant in the early stage of hydrolysis, prolonged reaction produced more glucose than cellobiose. Fraction I and fraction II-a acted synergically on Avicel. 13. Optimum pH for the activities of cellulase fraction I, fraction II-a, fraction II-b and fraction III were found to be 5.5, 5.0, 4.0 and $4.0{\sim}4.5$, respectively. These fractions were found to be stable in the range of pH $3.0{\sim}7.5$. 14. Optimum temperature for the activities of fraction I, fraction II-a, fraction II-b, and fraction III were $50^{\circ}C$, $55^{\circ}C$, $60^{\circ}C$ and $55^{\circ}C$, respectively. No less of activity was found by heating 120 minutes at $55^{\circ}C$ and fraction II-a was more stable than the others at $60^{\circ}C$. 15. Fraction I and fraction II-b were activated by $Ca^{++}$ and $Mg^{++}$ but inhibited by $Hg^{++}$ and $Ag^{+}$.

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Extraction and Purification of Ginseng Oligopeptides with Antilipolytic Activities (Antilipolytic Activity를 보유하는 인삼 Oligopeptide의 추출 및 정제)

  • Kim, Su-Ill;Na, Jee-Yeong;Jo, Do-Hyun;Lee, Chun-Yung
    • Applied Biological Chemistry
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    • v.30 no.1
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    • pp.88-94
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    • 1987
  • To investigate ginseng oligopeptides with biological activities, the water extract was purified by ultra-filtration, gel filtration, ion-exchange and thin layer chromatography. Ultra-filtered water extract exhibited antilipolytic activity, inhibiting epinephrine-induced lipolysis in the isolated fat cells of rat epididymal adipose tissue. The filtrate was separated into 3 fractions by Sephadex G-25 gel filtration. Peptides were found only in the first fraction(S-FI). Saponine and sugars were also detected in tie fraction. S-FI fraction resolved further into 6 fractions by Dowex 50 ion-exchange chromatography. The sugar and saponine depleted fraction(P-F2) from the second chromatography showed antilipolytic activity. The P-F2 fraction revealed 6 spots on TLC. The 6 spots were isolated by TLC and identified as peptides.

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Pharmacological Studies on Ether Fraction of Corni Fructus (산수유 에텔분획물의 약리작용에 관한 연구)

  • 이은방;최병천;조태순
    • YAKHAK HOEJI
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    • v.29 no.1
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    • pp.1-10
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    • 1985
  • The ether fraction obtained from dry fruit of Cornus officinalis was investigated for the anti-inflammatory action, acute toxicity and central nervous system activities. From several pharmacological examinations, it was found that the ether fraction suppressed considerably carrageenin edema at the dose of 600mg/kg p.o. in rats, inhibited the granuloma formation in rats as given 100mg/kg p.o. for 7 days and decreased the swelling of both of complete adjuvant injected and noninjected (contralateral) paws of the rats at the dose of 100mg/kg p.o. given for 14 days. The $LD_{50}$ of the fraction are estimated to be more than 2,000mg/kg p. o. and 642mg/kg i. p. in mice. And the fraction did not show any sedative, stimulative, analgesic and anticonvulsant action but exhibited hypothermic action. These results might be concluded that the ether fraction of Corni fructus showed anti-inflammatory action in both of acute and chronic type models without any considerable central nervous depressant activity and exhibited very weak acute toxicity in mice.

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Neuroprotective Effect of Ethyl Acetate Fraction of Portulaca oleracea L. (마치현 에틸아세테이트 분획물의 뇌세포 보호효과)

  • Im, Nam Kyung;Jeong, Gil Saeng
    • Korean Journal of Pharmacognosy
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    • v.44 no.4
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    • pp.379-383
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    • 2013
  • Portulaca oleracea L. is known to have many biological benefits such as anti-oxidant, anti-inflammatory, anti-allergic and anti-tumor. The objective of this study is to explore the neuroprotective effect of P. oleracea L. against glutamate-induced oxidative stress in mouse hippocampal HT22 cells. P. oleracea L. 70% ethanol extract and solvent fractions have the potent neroprotective effects on glutamate-induced nerotoxicity by induced the expression of heme oxygenase (HO)-1 in HT22 cells. Especially, ethyl acetate fraction showed higher protective effect. In HT22 cell, P. oleracea L. treatment with ERK inhibitor (PD98059) and c-JUN N-terminal kinase (JNK) inhibitor (SP600125) reduced P. oleracea L. ethyl acetate fraction induced HO-1 expression and P. oleracea L. ethyl acetate fraction also increased ERK and JNK phosphorylation. Furthermore, we found that treatment of P. oleracea L. caused the nuclear accumulation of Nrf2. In conclusion, the ethyl acetate fraction of 70% ethanol extract of P. oleracea L. significantly protect glutamate-induced oxidative damage by induction of HO-1 via Nrf2, ERK and JNK pathway in mouse hippocampal HT22. Taken together these finding suggest that P. oleracea L. ethyl acetate fraction is good source for taking active compounds and may be a potential therapeutic agent for brain disorder that induced by oxidative stress and neuronal damage.

Reduction of Allergenicity of Wheat Flour by Enzyme Hydrolysis (효소 분해에 의한 밀가루의 항원성 저감화)

  • Park, Ju-Yeon;Ahn, Jeung-Yeub;Hong, Hee-Ok;Hahn, Young-Sook
    • Korean Journal of Food Science and Technology
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    • v.36 no.1
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    • pp.152-157
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    • 2004
  • Gluten was extracted from domestic wheat flour using UTH buffer (4 M urea in 0.1 M Tris-HCl, pH 8.6) and validated by SDS-PAGE analysis for production of wheat flour products with reduced gluten content.. Anti-gluten polyclonal antibody was made by administering extracted gluten fraction on animal model. Anti-gluten serum titer of extracted gluten fraction was evaluated by ELISA, and that of antibody titer according to administration period. Anti-gluten sera were used for ELISA and immunoblot analysis before and after hydrolysis of gluten fraction at optimal pH and temperature condition for each protease. Gluten fraction separated by SDS-PAGE showed several bands covering 75 to 10 kDa, in which anti-gluten sera were 25, 34, and 45 kDa. Enzyme hydrolysis of gluten fraction revealed protein band sizes to be lower than 15 kDa. Content of pretense from bovine pancreas (b.p. protease) for gluten hydrolysis was estimated as 1 mg in 10 mL gluten fraction extracted for 4 hr.

Detection of Changes of the Population Fraction Nonconforming in the p Control Chart (p관리도의 불량률의 변화 탐지)

  • Chang, Kyung;yang, Moon-Hee
    • Journal of Korean Society for Quality Management
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    • v.25 no.3
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    • pp.74-85
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    • 1997
  • In this paper we calculate the subgroup size necessary for detecting the change of percent defective with several detection probabilities for orginal population fraction nonconforming p, changed population fraction nonconforming $p^*$, and the ratio k=$p^*$/p in the usage of p control charts. From our calculation we can know the error level of normal a, pp.oximation in detection probability calculation and recommend the subgroup size with lower error levels of normal a, pp.oximation, and then we show the reasonable subgroup size necessary for p, $p^*$, k, and the detection probability of the change of fraction nonconforming in a process. The information that we here show in tables will be useful when p control chart users decide the subgroup size in the p control chart users decide the subgroup size in the p control chart.

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Studies on the Naringinase Produced by Aspergillus nidulans (Part I) Purification and Characteristics of Naringinase (Aspergillus nidulans 가 생산하는 Naringinase 관한 연구 (제2보) Naringinase의 정제 및 물리화학적 성질)

  • 변유량;문순옥;유주현
    • Microbiology and Biotechnology Letters
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    • v.6 no.2
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    • pp.65-73
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    • 1978
  • Naringinase extracted from the culture media of Aspergillus nidulans was purified, and the activity was proved to be stronger by 781 fold in the part of precipitation with ammonium sulfate, and column chromatography using DEAE-Sephadex A-25 and Sephadex G-100. Two fractions which had the same enzyme activity were isolated by the purification. Both fractions showed the highest enzymes activity under the reaction conditions of pH 5.0 and 40$^{\circ}C$. Molecular weight of fraction I and fraction II were estimated as 78,000 and 26,000 respectively. This indicated that fraction I would be trimer of fraction II. The enzyme was inhibited by glucose and rhammose, and the Km value was calculated to be 2.3${\times}$ 10$\^$-6/ g/ml.

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Effect of Ginseng Saponin Fraction on Cleavage of pBR322 by Several Restriction Endonuclease (인삼 Saponin분획이 제한효소에 의한 pBR322 절단에 미치는 영향)

  • 강정훈;조영동
    • Journal of Ginseng Research
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    • v.9 no.2
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    • pp.240-247
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    • 1985
  • Attempts were made to see if we could cut more pBR322 in the presence of ginseng saponin fraction in connection with possibly for shortening the enzymatic reaction time and the amount of the enzyme to be used. The following results were obtained restriction endonucleases such as AccI, XhoII, SaII, and HincII were observed to cut pBR322 efficiently at 10-1% ginseng saponin fraction. In case of BamHI, 10-2% ginseng saponin fraction was observed to the most effective concentration. Such cumulative results suggest that ginseng saponin fraction would play important role as far as the cleavage of pBR322 for short period by endonucleases is concerned.

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Cytotoxic Effects of Hot Water Soluble Polysaccharides from Mushroom, Lentinus edodes and Vitamin A & E Supplementation against $P_388$ Cells (표고버섯의 열수추출 다당류 및 비타민 A와 E 첨가가 $P_388$의 세포독성에 미치는 영향)

  • 최미연
    • Journal of Nutrition and Health
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    • v.28 no.11
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    • pp.1091-1099
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    • 1995
  • The cyutotoxic effects of hot water soluble polysaccharides extract(PS) from the mushroom, Lentinus edodes, and in combinations with vitamin A or vitamin E on life span of ICR mice bearing P388 cancer cells and in vitro against P388 cancer cells were examined. The chemical components of PS and fractions were analyzed and survival time and cell number of P388 treated with extract fractions with and without vatamin A or E supplementation were also measured. The results obtained were summarized as follows; The extract of fraction B was shown to have the highest antitumor activity against P388 implanted in ICR mice. The antitumor fraction B was consisted of 82.0% of polysaccharide and 4.2% of protein. All three fractions seemed to have in vivo antitumor activity against P388, and fraction B showed the highest activity, In vitro P388 cell growht was inhibited 76%, 89%, 54% by the addition of fraction A, B and C respectively. Vitamin A or E did not appear to have any accelerating effects on either in vivo or in vitro cell cytotoxicity when each of them was combined with the PS and fractions. All three fractions contained more than 68% of polysaccharides. The fraction B showed the highest value of 88% in polysaccharides. Monsaccharides of the fraction B were identified as galactose(59.1%), glucose(29.2%), fructose(2.8%) and uronic acid(4.2%). Hydrolysis of protein from the fraction B was didentifed to have 17 kinds of defined and 5 undifined amino acids. The inhibitory effects of the hot water extracts from mushroom against cancer cell growth of P388 were stronger than the control group. And the survival time of ICR mice was shown to be 161% between the control group and the experimental groups.

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Antimutagenic Effect of Bacillus natto Isolated from Natto

  • Yun, Soon-Il
    • Journal of Applied Biological Chemistry
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    • v.48 no.3
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    • pp.133-137
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    • 2005
  • In vitro antimutagenicity of Bacillus natto islolated from Natto, Japanese traditional fermented food, was investigated using umu-test. Mutagenicity of S9-activated metabolites of Trp-P-2 and IQ for Salmonella typhimurium TA 1535/pSK1002 was remarkably inhibited by addition of bacterial cells and their cytoplasmic fraction. Desmutagenicity by cytoplasmic fraction increased with increasing concentration of the fraction. Bioantimutagenic effect of cytoplasm on Salmonella typhimurium SD-100 did not show bioantimutagenic activity against mutated bacterial cells induced by Trp-P-2. Cytoplasmic fraction exhibited 17% bioantimutagenicity due to desmutation caused by IQ.