• Title/Summary/Keyword: aspergillus

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Detection of Aspergillus and Penicillium genera by Enzyme-Linked Immunosorbent Assay Using a Monoclonal Antibody

  • Kwak, Bo-Yeon;Shon, Dong-Hwa;Kwon, Byung-Joon;Kweon, Chang-Hee;Lee, Ke-Ho
    • Journal of Microbiology and Biotechnology
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    • v.11 no.1
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    • pp.21-28
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    • 2001
  • Enzyme linked-immunosorbent assay (ELISA) for a rapid detection of fungi, Aspergillus and Penicillium genera in food, were developed and their efficiencies were approved by detecting artificially contaminated agricultural commodities. Mice were immunized with partially purified Aspergillus flavus extracellualr polysaccharide (EPS) and lymph node cells of the mice were fused with the myeloma cells for production of monoclonal antibodies. Mab 1G11, one of the antibodies, was selected and purified. A sandwich ELISA was established and its detection limit toward A. flavus EPS was 1mg/ml. Among the 59 strains tested (including 18 species of Aspergillus, 16 of Penicillium, 11 of Fusarium, 1 of Absidia, 2 of Alternaria, 2 of Candida, 2 of Cladosporium, 2 of Geotrichum, 2 of Mucor, 2 of Rhizopus, 1 of Trichoderma), species of Aspergillus and penicillium had a high reactivity with Mab 1G11 even up to 10,000 times dilution of culture broths. The other genera except Cladosporium resinae showed no reactivity, thus Mab 1G11 was specific to the genera of Aspergillus and Penicillium. The epitope of A. flavus EPS against monoclonal Mab 1G11 was on the carbohydrate moiety when 1 to 100$\mu g/g$ A. flavus EPS were put into rice, potato, and mandarin orange, the average recoveries detected by sandwich ELIA were 123, 59, and 76%, respectively. Correlation was found to be linear between the EPS, and mycelium of A. flavus and Penicillium citrinum grown in a liquid medium (r=0.87 and 0.96), and also between the EPS and colony forming unit in solid media of rice of potato (r=0.91-0.99).

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Intergeneric Protoplast Fusion between Rhizopus oryzae and Aspergillus oryzae (Rhizopus oryzae와 Aspergillus oryzae의 속간 원형질체융합)

  • Lee, Soo-Youn;Jung, Sung-Won;Kim, Seong-Han;Lee, Yung-Nok
    • Korean Journal of Microbiology
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    • v.31 no.3
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    • pp.218-223
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    • 1993
  • Conditions for the release and regeneration of protoplasts form Rhizopus oryzae and intergeneric protoplast fusion between Rhizopus oryzae and Aspergillus oryzae were studied. High yields of protoplast fusion between Rhizopus oryzae and Aspergillus oxyzae were studied. High yield of protoplasts from young germilings of R. oryzae were obtained by using lytic enzymes containing chitosanase (3 mg/ml), chitinase (3 mg/ml) and Novozym 234 (5 mg/ml). 0.5M glucose was used as the osmotic stabilizer and optimum pH of buffer was determined to be pH 7.5-8.0. Under these conditions, protoplasts were formed after about 3-4 hrs incubation. Approximately, 1.0%-4.9% of these protoplasts were formed after about 3-4 hrs incubation. Approximately, 1.0%-4.9% of these protoplasts regenerated on solid medium with a soft agar overlay. We have also carried out protoplasts fusion between R. oryzae and A. oryzae and have succeeded in obtaining three types of intergeneric fusants. In these experiments, 35% PEG-4000 and 10 mM CaCl$_{2}$ were used as fsogenic agents, and auxotrophic properties were used as a genetic marker to select fusants. Complementation frequency be protoplasts fusion of A. oxyzae and R. oryzae was 4.4% * 10$^{-5}$ . The fusant strains of the first type were prototrophs showing an Aspergillus type morphology with dark-yellow sporulation, those of the second type were also Apergillus type morphology but showed no sporulation. And the strains of the third type stopped growing when fusion products grown on regeneration minimal medium were transferred to fresh minimal medium. The formation of fusion products was observed by fluorescent vital stains for complementary labelling of protoplats from R. oryzae and A. oryzae. Rhodamine 6G and fluorescein diacetate wer useful complementary vital stains of Rhizopus and Aspergillus protoplasts for visualization of requency and type (dicell, multicell) of fusion.

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Solubilization of Insoluble Phosphates by Aspergillus sp. PS-104 Isolated from Soil (토양에서 분리한 Aspergillus sp. PS-104 균주에 의한 난용성 인산염 분해)

  • Kang, Sun-Chul;Shin, Seung-Yong
    • Korean Journal of Environmental Agriculture
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    • v.26 no.1
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    • pp.36-41
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    • 2007
  • Phosphate-solubilizing microorganisms were isolated from soil around Kyungnam and Kyungbook regions using potato dextrose agar-calcium phosphate medium. A fungus with the greatest phosphate-solubilizing activity was selected and identified to Aspergillus sp. PS-104, based on the morphological characteristics of conidiophore and conidia; unbranching type of conidiophore, terminally swelling of conidiophore and septate of mycelium, in malt extract agar and potato dextrose agar media. The optimum temperature and initial pH to solubilize rock phosphate in potato dextrose broth-rock phosphate medium were $30^{\circ}C$ and pH 7.0, respectively. In these optimum conditions, phosphate-solubilizing activities of Aspergillus sp. PS-104 against four twos of insoluble phosphate, tricalcium phosphate, aluminium phosphate, hydroxyapatite and rock phosphate, were quantitatively determined. As result, the maximum phosphate-solubilizing activity was obtained with tricalcium-phosphate (1,900 ppm) while minimum activity was obtained with hydroxyapatite (320 ppm). Futhermore, phosphate-solubilizing activity of Aspergillus sp. PS-104 was found higher when treated with nitrates as compared to the ammonium salts as a nitrogen sources.

Characterization of Xylanase from an Hybird between Aspergillus oryzae var. oryzae and Aspergillus Nidulans 514 by Nuclear Transfer (핵전이에 의한 Aspergillus oryzae var. Oryzae와 Aspergillus nidulans 514의 잡종으로부터 생산된 Xylanase의 특성)

  • Yang, Young-Ki;Moon, Myeng-Nim;Park, Hyung-Nam;Lim, Chae-Young
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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    • pp.50-58
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    • 1996
  • Interspecific hybrids between Aspergillus oryzae var oryzae and A. nidulans 514 were obtained by nuclear transfer technique. Several autotrophic mutants isolated from conidiospores of the two strains were mutagenized with ultraviolet and N-methyl-N-nitrosoguanidine. Optimal conditions for formation of intergeneric hybrids were investigated. Frequencies of hybrid formation by nuclear transfer were $3{\times}10^{-5}{\sim}1{\times}10^{-5}$. From observation of genetic stability, conidial size, DNA content, and nuclear stain, it was suggested that their karyptypes are aneuploid. The hybrids showed 1.1~1.4 fold higher xylanase activities than parental strains did. The xylanase of Aspergiilus sp. TAVD514-3 was purified and some of it's enzymatc characteristics were investigated. The enzyme was purified about 85 fold with an overall yield of 17% from the culture medium by ammonium sulfate fractionation, Sephadex G-75 gel permeation chromatography, and CM-sephadex A-50 ion exchange chromatography. The purified enzyme functions optimally at pH 9.0 and 80$^{\circ}C$. The enzymatic activity was increased by the presence of $Mg^{2+}$ and $Mn^2$ ions.

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Formation and Fusion of Protoplasts from the Cellulolytic Fungi, Aspergillus niger MAN-831 and Aspergillus wentii MAW-538 (Cellulase를 생산하는 Aspergillus niger MAN-831과 Aspergillus wentii MAW-538의 원형질체 형성 및 융합)

  • 박석규;이상원;문일식;손봉수;강성구
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.6
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    • pp.964-969
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    • 1995
  • For the effective utilization of cellulosic biomass, conidial protoplast fusion between Aspergillus niger MAN-831(${\beta}-glucosidase$) and A. wentii MAW-538(CMCase and avicelase), which produced potently cellulolytic enzymes was carried out. Optimal conditions for formation and regeneration of protoplast were conidiospore age-5 dyuas. $2-DG-30\mu\textrm{g}/ml$, preincubation time-4 hours, osmotic stabilizer-0.7M KCl, novozyme(7mg/ml)+driselase(2.5mg/ml) and reaction time of enzyme-5 hours. Optimal conditions for protoplast fusion were obtained by treatment of protoplasts with 15mM CaCl2 and 25% polyethylene glycol 4000(pH 6~7) as fusogenic agent at $36^{\circ}C$ for 25~30 minutes. The frequency was then $7.94{\times}10^{-4}$. CMCase, avicelase and ${\beta}-glucosidase$ activity of fusant F-208 strain was 1.5, 1.3, 1.2 times higher than those of parental strains, respectively.

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Effect of Meju Shapes and Strains on the Chemical Composition of Soybean Paste (Bacillus속과 Aspergillus oryzae로 만든 메주가 개량식 된장의 성분에 미치는 영향)

  • Seo, Jeong-Sook;Man, Eun-Mi;Lee, Taik-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.15 no.4
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    • pp.1-9
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    • 1986
  • The mashes of soybean paste were preparea using the conventional meju fermented naturally by wild microoganisms or the new types of meju fermented by pure cultures of Aspergillus oryzae, Bacillus natto and B. subtilis to elucidate changes during the aging period. The results obtained are as follows ; The soybean paste made with conventional meiu and Asp. oryzae meju showed higher content of amino nitrogen than those of B, natto and B. subtilis meju. Soybean paste made with conventional meju contained a little more content of total and reducing sugars than other soybean pastes. ph during aging period was higher than 5.0 for the Asp. oryzae paste while less than 4.5 for B. subtilis paste. Aspartic acid. threonine, serine, glutamic acid, glycine, alanine, cystine, valine, methionne, leucine and histidine for Asp. oryzae paste ; tyrosine, arginine and proline for conventional meju paste; and isoleucine and phenylalanine for B. subtilis paste were found to be peak amount 90 days after the preparation. The content of total free amino acid was high in the order of Asp. oryzae paste, conventional paste, B. natto paste and B. subtilis paste.

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Characterization of the Bacterial Cell Wall Lytic Enzyme Produced by Aspergillus sp. HCLF-4 (Aspergillus sp. HCLF-4에 의해 생성되는 세균세포벽 분해효소의 특성)

  • 임진하;민병례;최영길
    • Korean Journal of Microbiology
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    • v.37 no.1
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    • pp.15-20
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    • 2001
  • In this study, we have isolated bacterial cell wall lytic enzyme in the culture supernatant of Aspergillus sp. HCLF-4. This hydrolase showed cell wall lytic activity against Anabaena cylindrica. The extracellular enzyme was produced by Aspergillus sp. HCLF-4 when it was grown in a PDB media containing 0.05% heat killed Micrococcus luteus cells. The molecular weight of lytic enzyme was about 14.3 kDa. The optimal pH and temperature for the activity of this enzyme were 3.0~4.0 and $30^{\circ}C$, respectively. This hydrolase activity was reduced by $Na^{+}$, $Li^{+}$, $Ca^{2+}$, $Cu^{2+}$, $Fe^{3+}$, EDTA, and PMSF, whereas it was increased by $Mg^{2+}$, $Mn^{2+}$>. The enzyme has N-acetylmuramyl-L-amidase or endopeptidase activity.

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Physicochemical Characteristics of the Mash Quality of Cheongju Prepared Using Different Nuruks (누룩종류를 달리한 청주 술덧의 이화학적 특성)

  • Jeong, Myeong Eun;Kang, Soon Ah;Cheong, Chul
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.15 no.8
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    • pp.5150-5158
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    • 2014
  • This study examined the physicochemical characteristics of mashing prepared using different Nuruks (traditional Nuruk, Nuruk added Aspergillus oryzae, Nuruk added Aspergillus kawachii, and Nuruk added Aspergillus oryzae and Aspergillus kawachii), and to obtain the basic data for Korean Cheongju production. The four different mashes were compared for their brewing characteristics and indicated a normal fermentation pattern. The Nuruk made from A. oryzae or A. kawachii showed higher fermentation ability than the other samples tested. In addition, the analyzed results of fermented mashing showed that the Nuruk made from A. oryzae or A. kawachii had a significantly higher aroma profile, such as higher alcohol and ester than the other samples tested, whereas the Nuruk made from a mixture of A. oryzae and A. kawachii had a much higher organic acid content. In conclusion, the Nuruk prepared using A. oryzae or A. kawachii had a positive effect on the complete fermentation and enhanced aroma compounds for Korean Cheongju production.

Fungal Microflora on Korean Home-made Meju (재래식(在來式) 메주에 분포(分布)하고 있는 진균(眞菌)에 관한 조사(調査) 연구(硏究))

  • Park, Kyoung-Ja;Kim, Young-Mi;Lee, Bae-Ham;Lee, Bok-Kwon
    • The Korean Journal of Mycology
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    • v.5 no.1
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    • pp.7-12
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    • 1977
  • For this study 3 samples of the home-made meju and 3 samples of the improved meju were selected according to their characteristics. A total of 25 strains of true fungi were isolated from the samples of the home-made meju and identified by the Key of Alexopoulos and Raper, using a strain Aspergillus oryzae A-113 as a control. Amylolytic and proteolytic enzyme activities of the isolated strains were investigated ana the results obtained were as follows. 1. The 25 strains from the home-made meju were identified into 2 Aspergillus oryzae, 14 Asp. flavus, 6 Penicillum spp. 1 Candida sp 1 Spicaria sp and 1 Rhizopus sp. 2. The 3 strains from the improved meju were all identified as Aspergillus oryzae. 3. Aspergillus flavus, A-B, from the home-made meju was found to he the strongest strain in ${\alpha}-amylase$ activity and also to be similar to the strains of Aspergillus orzae from improved meju. 4. Aspergillus flavus, A-7, from the home-made meju was found to be the strogest strain in ${\beta}-amylase$ activity and stronger than that from the improved meju. 5. Aspergillus flavus B-3, was found to be the strongest strain in protease activity and stronger than that from the improved meju. 6. Some of the strains from the home-made meju turned out to be harmful strains, such as Penicillium spp. which secrete antibiotics, Asp. flavus which secretes mycotoxin, Candida sp which causes skin diseases, Spicaria sp. which is a insect pathogen. 7. Rhizopus sp was also found but it has not been proved to be harmful.

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