• Title/Summary/Keyword: Aspergillus sp.

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Characteristics of the alkaline protease from the moderate halophile, Halomonas sp. ES 10 (Halomonas sp. ES 10이 생산하는 alkaline protease의 특성)

  • Kim, Chan-Jo;Oh, Man-Jin;Choi, Seong-Hyun
    • Applied Biological Chemistry
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    • v.35 no.4
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    • pp.237-241
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    • 1992
  • The protease from Halomonas sp. ES 10 was purified by methanol precipitation, gel filtration on Sephadex G-150 and G-200, and ion exchange chromatography on DEAE-Sephadex A-50. The purified enzyme was found to be homogeneous by polyacrylamide gel electrophoresis. The specific activity of purified enzyme was 1,014 units/mg protein, and the yield of the total activity from the culture filtrate was 7%. The optimal temperature and pH for the enzyme activity were $35^{\circ}C$, and pH 11.0, respectively. And the enzyme was stable in the range of $pH\;7.5{\sim}11.0$. The residual activity of the enzyme was 70%, when the enzyme was incubated at $50^{\circ}C$ for 40 min. The Km value of the enzyme was 7.4 mg/ml to milk casein. $Li^+$, $Ca^{2+}$, SDS and Tween 80 were appeared to activators, whereas $Hg^{2+}$ and EDTA to inhibitors. The addition of DFP and PMSF showed the relative enzyme activities of 63% and 107%, respectively, suggesting that the enzyme may not belong to serine type protease. When the alkaline protease was treated with 0.5 M and 1 M NaCl, the relative enzyme activities were 95% and 65%, respectively. This enzyme showed 20% and 15% higher enzyme activity than that of Aspergillus oryzae (Sigma Chemical Company product, P4755) in the presence of 0.5 M and 1 M NaCl.

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Purification and Biological Characterization of Wild-type and Mutants of a Levan Fructotransferase from Microbacterium sp. AL-210 (Microbacterium sp. A-210이 생성하는 Levan fructotransferase의 정제 및 생물학적 특성에 관한 연구)

  • Hwang, Eun-Young;Jeong, Mi-Suk;Cha, Jae-Ho;Jang, Se-Bok
    • Journal of Life Science
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    • v.19 no.9
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    • pp.1218-1225
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    • 2009
  • Difractose anhydrides (DFAs) is studied as a sweetener for diabetics because of its structural property. DFAs have four types: DFA I, III, IV (degradation of levan) and V (degradation of inulin). Especially, DFA IV has been shown to enhance the absorption of calcium in experiments using rats. Levan fructotransferase is an enzyme for producing di-d-fructose-2,6':6,2-dianhydride (DFA IV). To identify structural characterization, we purified wild-type and mutants (D63A, D195N and N85S) of levan fructotransferase (LFTase) from Microbacterium sp. AL-210. These proteins were purified to apparent homogeneity by Ni-NTA affinity column, Q-sepharose ion exchange and gel filtration chromatography and detected by SDS-PAGE. They were also analyzed by circular dichroism (CD) measurements, JNET secondary structure prediction, activity measurements at various temperatures, and pH analysis. The optimum pH for the enzyme-catalyzed reaction was pH 7.5 and optimum temperature was observed at $55^{\circ}C$. Along with wild-type LFTase, mutants were analyzed by CD measurement, fluorescence analysis and differential scanning calorimetry (DSC). N85S showed less $\alpha$-helix and more $\beta$ strand than others. Also, N85S showed almost the same curve as wild-type in their steady-state fluorescence spectra, whereas mutant D63A and D195N showed higher intensity than wild-type. The amino acid sequence of wild-type LFTase was compared to the sequences of exo-inulinase from Aspergillus awamori, a plant fructan 1-exohydrolase from Cichorium intybus, and Thermotogo maritime (Tm) invertase and showed a high identity with Exo-inulinase from Aspergillus awamori.

A Study on the Qualitative Properties of Traditional Sake Using Allbanggae (올방개(Eleocharis kuroguwai Ohwi)를 이용한 전통약주의 품질특성에 관한 연구)

  • Cheong, Chul;Rhee, In-Sook;Lee, See-Kyung;Kang, Soon-Ah
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.6
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    • pp.784-791
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    • 2008
  • A new sake fermented using a mixture of Aspergillus sp., Rhizopus sp. and yeast (Saccharomyces cerevisiae) containing allbanggae was developed in this study. The effects of the allbanggae contents ($0{\sim}50%$) in the sake on the qualitative and sensory properties and flavor components were investigated. The fermentation with allbanggae, especially in sake containing 10% allbanggae, showed the highest level of alcohol ($15.3{\sim}16.4%$), whereas the amount of alcohol decreased with increasing allbanggae contents. Amino acidity in sake containing 10% and 20% allbanggae indicated the highest levels ($0.90{\sim}1.20%$). In addition, it was observed that the sake containing 10% allbanggae showed the smallest level of reducing sugars, which were increased with increasing allbanggae ratio. Furthermore, various organic acids in the sake were detected such as citric, succinic, malic and acetic acids, among which the acetic acid showed the highest amount ($160.3{\pm}8.0{\sim}253.3{\pm}20.3mg/100mL$). The major amino acids detected in the sakes were alanine, proline, histidine, phenylalanine, lysine and glutamic acid. The sake containing 20% allbanggae indicated the highest amino acids amount among the allbanggae ratios. Based on these results, it was concluded that the sake containing 20% allbanggae could be successfully commercialized.

6- [(Halopheny1) amino]-7-Bromo-5,8-Quinolinediones Treatment of Candidiasis in Normal Mice

  • Kim, Sung-Hee;Kim, Do-Hee;Lee, In-Kyung;Kim, Dong-Hyun;Ryu, Chung-Kyu
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.167-167
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    • 1996
  • A series of 6-[(N-Halophenyl)amino] -7-Bromo-5,8-quinolinediones (BQs) newly synthesized were tested for antifungal activities, in vitro, against Candida sp., Aspergillus niger and Trichophyton mentagrophytes. The MIC values were determined by the twofold dilution method. Among these derivatives, BQ4 and 7 showed more potent antifungal activities than fluconazole and ketoconazole. BQ 4 and 7 were compared with fluconazole in the treatment of established systemic infections with Candida albicans in normal rats. Intraperitoneally administered BQs for 7 days and 14 days reduced Candida albicans colony count in kidneys and livers as well as fluconazole. The therapeutic potential of BQs has been assessed by evaluating their activities (survival rate) against systemic infections in normal mice with Candida albicans. These compounds, particularly showed activities comparable with fluconazole. BQ4 and 7 had about EO$\_$50/, 0.05, 0.06mg/kg but fluconazole had ED$\_$50/, 6.0mg/kg respectively.

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Synthesis of 6- (Arylamino) -7-Alkylthio -5,8-Quinolinediones for Evaluation of Antifungal Activities

  • Kim, Sung-Hee;Kim, Do-Hee;Lee, In-Kyung;Kwon, Sang-Mee;Jung, Sung-Hee;Ryu, Chung-Kyu
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.166-166
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    • 1996
  • A series of 6-(N-arylamino)-7-alkylthio-5,8-quinolinedione derivatives(SQ1-12) were newly synthesized for the evaluation of antifungal activities. 6-(N-Arylamino)-7-chloro-5,8-quinolinediones (RCKs) were treated with Na$_2$S/(CH$_3$)$_2$SO$_2$ in EtOH In give SQs. RCKs were prepared by regioselective nucleophilic substitution of 6,7-dichloro-5,8-quinolinediones with arylamines. In the presence of CeCl$_3$, the N-arylamino groups were introduced at the 6-position of 5,8-quinolinedione ring by the regioselective substitution. These derivatives 1-12 were tested for antifungal and also antibacterial activities, in vitro, against Candida sp., Aspergillus niger and Trichophyton mentagrophytes. The MIC values were determined by the two-fold dilution method.

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Actinobacteria from Cow Feces: Isolation, Identification and Screening for Industrially Important Secondary Metabolites

  • Semwal, Preeti;Rawat, Vinay;Sharma, Pushpendra;Baunthiyal, Mamta
    • Microbiology and Biotechnology Letters
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    • v.46 no.1
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    • pp.68-76
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    • 2018
  • Actinobacterial strains isolated from Cow feces were studied for their antifungal attributes against phytopathogens and industrially important enzymes. A total of 30 Actinobacterial strains were obtained from 10 samples of cow feces. All the strains were belonging to the genera Streptomyces on the basis of morphological and chemotaxonomic analysis. During preliminary screening, out of 30 strains, 15 strains (50%) showed antifungal activity against five fungal phytopathogens including Aspergillus niger, Fusarium solani, Fusarium oxysporum, Macrophomina phaseolina and Rhizoctonia solani. While, isolate GBTCF-26 was found to be most active against R. solani with 62.2% inhibition of fungal mycelium, GBTCF-09 was prominent against F. solani and F. oxysporum with percent inhibition of 61.1% and 58.8%, respectively. Out of 30 strains, 19 (63.3%), 16 (53.3%), 11 (36.7%), 10 (33.3%), 4 (13.3%) and 8 (26.7%) strains were producing amylase, caseinase, gelatinase, lipase, chitinase and cellulose, respectively. The selected strains, GBTCF-09, GBTCF-21 and GBTCF-26, were identified as Streptomyces sp. on the basis of their 16S rDNA sequence. The study supports the idea that the Actinobacteria from unique niches (Cow feces) possess the production potential of industrially important enzymes including bioactive molecules.

Studies on Changes of Aflatoxin Productivity and Properties by a Pseudomonad (녹농균과 진균류의 생물학적 상호작용에 의한 Aflatoxin 류의 생성능과 성질 변화에 대한 연구)

  • Lee, Young-Nam;Kim, Joo-Deuk
    • The Korean Journal of Mycology
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    • v.8 no.1
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    • pp.59-62
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    • 1980
  • Investigation on modification of aflatoxin structures by Pseudomonads was attempted as a biological detoxifiying process of mycotoxins. Firstly, when any variation of aflatoxin yield of Aspergillus parasiticus in a mixed culture with Pseudomonas aeruginosa was examined, there was no noticible effect by growth of Pseudomonads on aflatoxin yield of Asperillus sp. Secondly, when capacity of Pseudomonas aeruginosa utilizing aflatoxin as a carbon source for its growth was tested, there was some indication that aflatoxin might be used for growth of Pseudomonads. It was also noticed that the residual aflatoxin showed different migrating pattern compared with that of the intact aflatoxin by thin layer chromatography. Thirdly, the cell-free extract prepared from Pseudomonas aeruginosa grown in a glucose minimal medium supplemented with aflatoxin and the intact aflatoxins were incubated in the presence of $Mg^{++}$. After a certain length of incubation period, the reaction mixtures were applied on TLC plates. As a result, aflatoxins in the reaction mixture migrated differently as the control did. Such results may indicate that some changes of aflatoxin properties were induced by Pseudomonads.

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Fungal-Sporulation Suppressing Substances Produced by Pseudomonas aeruginosa KMCS-1

  • Min, Bu-Yong;Shim, Jae-Young;Kim, Kun-Woo;Lee, Jong-Kyu;Choi, Hyung-Tae;Yoon, Kwon-Sang
    • Journal of Microbiology
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    • v.34 no.3
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    • pp.284-288
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    • 1996
  • Among the bacteria isolated from compost piles of cattle excretion in a pasture located at the suburbs of Chunchon city, Pseudomonas aeruginosa KMCS-1 was selected for the test of antifungal substances produced. Six fractions were separated by silica gel column chromatography, and then the antifungal activity of each fraction was assayed against Escherichia coli, Bacillus subtilis, Candida albicans, Rhizopus sp., Aspergillus nidulans, Coprinus cinereus, and Pyricularia oryzae by paper disc method. Two fractions showed significant suppressive activities against A. nidulans, C. cinereus, and P. oryzae however, their mycelial growth was not affected by neither of these fractions. Inhibitory activities of these fractions to sporulation was assayed at the concentration of 50. 25, 12. 5, and 6.25 $\mu$g/ml and the average inhibition rates against sporulation of A. nidulans, C. cinereus, and P. oryzae were 94.0, 98.3, and 77.9%, respectively. Further purification and analysis of active substances are now being conducted.

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Changes in microorganisms and Min Components during Takju Brewing by a Modified nuruk (개량누룩에 의한 탁주양조중 미생물과 중요성분 변화)

  • 이명숙
    • The Korean Journal of Food And Nutrition
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    • v.12 no.3
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    • pp.226-232
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    • 1999
  • To evaluate the characteristics of modified nuruk made by inoculation and cultivation of Rhizopus jap-onicus T2, Aspergillus oryzae L2 and hansenula sp. BC26 three different Takju mashes were made with modified Nuruk commercial Nuruk and rice koji and the changes in microorganisms and major compon-ents of mashes were investigated uring brewing. The numbers of yeast kept higher in the mash of mod-ified Nuruk or rice koji. The numbers of lactic acid bacteria were high in mash of commercial Nuruk but those were not in mash of modified Nuruk or rice koji. all mashes showed stable pH in a day and the pH kept higher in mash of modified Nuruk than in that of commer-cial Nuruk or rice koji. Increase in the content of total acid was moderate in mash of modified Nuruk or rice koji but it was too much in that of commercial Nuruk. increase in the content of amino acid was much highr in mash of modified Nuruk or commercial Nuruk than in that of rice koji. Increase in the content of alcohol was more rapid in mash of modified nuruk or rice koji than in that of commercial Nur-uk. The content of reducing sugar kept highter in mash of commercial Nuruk or modified Nuruk than in that of rice koji. A small amount of modified Nuruk accomplished favorable fermentation showing nor-mal patterns in microbiological and physicochemical changes during brewing.

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Characteristics of a Modified Nuruk Made by Inoculation of Traditional Nuruk Microorganisms (전통 누룩미생물들로 제조한 개량누룩의 특성)

  • 소명환
    • The Korean Journal of Food And Nutrition
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    • v.12 no.3
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    • pp.219-225
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    • 1999
  • In order to improve the quality of nuruk a Korean traditional fermenting agent for brewing nuruk was modified by a new method inoculating Rhizopus japonicus T2 Aspergillus oryzae L2 and hansenula sp. BC26 which had been isolated from traditional Nuruk. And the characteristics of modified Nuruk were investimgated as compared with current fermenting agents such as commercial Nuruk and rice koji of As-pergillus kawachii. The odor of modified Nuruk was savory but those of commercial Nuruk and rice koji were fetid andinodorous repectively. The extracted waters of modified Nuruk commercial Nuruk and rice koji were yellow pale yellow and colorless and showed 6.15, 6.01 and 3.30 of pH respectively. Mod-ified Nuruk had 7.6${\times}$106CFU/g of yeast but commercial uruk and rice koji had no yeast. Commercial Nuruk had 1.0${\times}$102CFU/g of lactic acid bacteria but modified Nuruk and rice koji had no lactic acid bacteria. The amylolytic and proteolytic activities of modified Nuruk were much higher than those of commercial Nuruk or rice koji. Seed mash of modified Nuruk had ester aroma but that of commercial Nuruk rather offensive odor and that of rice koji neither ester aroma or offensive odor. It seemed that if the modified Nuruk is used in seed mashing the supplement of acidulant is need to lower pH. The quality of modified Nuruk was thought to be much better than that of commercial Nuruk or rice koji.

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