• 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).

• The Korean Journal of Mycology
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• v.13 no.4
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• pp.243-247
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• 1985

Experiments were conducted to study effects of steroidal carbamate derivatives upon mycelial growth and aflatoxin production by Aspergillus flavus ATCC 15517. The basal medium was supplemented with various concentrations of these compounds and inoculated with spores. The developing cultures were incubated for 11 days at $28^{\circ}C$ without agitation. Aflatoxins were extracted with chloroform, separated by thin layer chromatography, and quantitated by ultraviolet spectrophotometry. At a concentration of 50 mg per 30 ml of medium., stigmasteryl-N-(2-chloroethyl) carbamate, cholesteryl- N - (2-chloroethyl) carbamate, $5{\alpha}-cholestan-3-one-oximino-N-(2-chloroethyl)$ carbamate and ${\beta}-sitosteryl-N-(2-chloroethyl)$ carbamate were the most effective in reducing aflatoxin production by Aspergillus flavus. However, cholest-4-ene-3-one-oximino-N-(2-chloroethyl) carbamate, at a concentration of 100 mg per 30 ml, significantly decreased aflatoxin production. There was no significant inhibition of mycelial growth by the addition of the various concentrations of these compounds.

• Mycobiology
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• v.31 no.4
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• pp.209-213
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• 2003

Aspergillus flavus K-03 isolated from poultry forming soil in Korea was studied for its ability to produce extracellular proteases on basal medium containing 2%(w/v) chicken feathers. The fungus was observed to be a potent producer of such enzymes. Keratinolytic enzyme secretion was the best at 15 days of incubation period at pH 9 and temperature $40^{\circ}C$. No relationship existed between the enzyme yield and increase of biomass. Enzyme production was suppressed by exogenous sugars in descending order arabinose>maltose>mannose>fructose. But glucose did not influence the enzyme activity. The keratinolytic enzyme released by the fungus demonstrated the ability to decompose keratin substrates as chicken feather when exogenous glucose was present. The keratinolytic activity was inhibited by $HgCl_2$ and serine-protease inhibitors such as phenymethylsulfonyl fluoride(100%), chymostain(88%), crystalline soybean trypsin inhibtor(80%), antipain(45%) and aprotinin(40%), and was not by cystein-protease and aspartyl-protease inhibitors. The enzyme activity is only partially inhibited by metallo-protease inhibitor. Thus, the enzyme secreted by A. flavus K-03 belongs to the alkaline serine-type protease.

• Journal of Microbiology and Biotechnology
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• v.13 no.2
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• pp.269-275
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• 2003

Chitosan-degrading activity induced by chitosan was founf in culture filtrate of Aspergillus flavus IAM2044. Aspergillus flavus IAM2044 had a higher level of chitosanolytic activity when chitosan was used as a carbon source, and yeast extract and peptone were supplemented as nitrogen sources. One of the chitosan-degrading enzymes was purified to homogeneity by ammonium sulfate precipitation followed by cation-exchange and gel filtration chromatographies. The enzyme was monomeric, and its molecular mass was 45 kDa. The optimum pH and temperature of the enzyme were 5.0 and $50^{\circ}C$, respectively. The activity was stable in the pH range of 3.5 to 7.0 and at a temperature below $50^{\circ}C$. Reaction products analyzed by the viscosimetric assay and thin layer chromatography clearly indicated that the enzyme was an exe-type chitosanase, $exo-{\beta}-D-glucosaminidase$, that released GlcN from the nonreducing ends of the oligosaccharide chains.

• Mycobiology
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• v.33 no.2
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• pp.77-83
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• 2005

Seventy-three fungal species belonging to forty-three genera were isolated from 40 samples of Saccharrum officinarum (collected from Naage-Hamadi canal in Qena Governorate, Egypt). Aspergillus, Trichoderma, Mucor and Pythium were the most common genera on the two isolation media. The dominant species of Aspergillus were A. niger, A. flavus, A. ustus, A. terreus and A. wentii. Some species were dominant on 40 g/l sucrose such as Aspergillus niger, A. flavus, Emericella nidulans, Trichoderma viride, Torula herbarum and Mamaria echinoeotryoides, while the dominant species on 10 g/l glucose were Mucor circinelloides, Aspergillus niger, Torula herbarum and Trichoderma viride. Mycotoxins including aflatoxins $B_1,\;B_2,\;G_1\;and\;G_2$, zearalenone and diacetoxyscirpenol were detected in the examined samples of Saccharrum officinarum. The mycelial growth of A. flavus, A. niger, Fusarium moniliforme and Torula herbarum decreased with the increase in Dimethoate concentrations, although 25 ppm was less effective than the higher levels of the insecticide ($75{\sim}200\;ppm$). Dimethoate stimulated the activity of Go-Tin A. niger, F. moniliforme and T. harbarum, while the Go-T activity was inhibited in A. flavus with the Dimethoate treatments.

• Korean Journal of Pharmacognosy
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• v.10 no.3
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• pp.112-118
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• 1979

21 herbal drugs registered in K.P. III were tested for contamination of fungi and isolation of aflatoxin producible strains. Initially contaminated fungi were Aspergillus group (41.28%) and Penicillium (47.26%) and the other fungi were contaminated somewhat. The most frequent isolation of Aspergillus group was Cnidii Rhizoma and that of Penicillium was Piperis Fructus nigri. Cnidii Rhizoma was the most contaminated drug and Cassiae Cortex was the least among them. Aspergillus flavus was isolated from 10 samples and Aspergillus parasiticus was detected in Glycyrrhizae Radix, Phellodendri Cortex. Aspergillus ochraceus was isolated from only Scutelariae Radix, and Fusarium nivale was isolated from Cnidii Rhizoma and Torreya Semen. None of Aspergillus and Penicillium was detected in only Coptidis Rhizoma. No strains of Aspergillus flavus and Aspergillus parasiticus isolated from were produced aflatoxin.

• Mycobiology
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• v.35 no.4
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• pp.219-225
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• 2007

A keratinolytic enzyme secreted by Aspergillus flavus K-03 cultured in feather meal basal medium (FMBM) containing 2% (w/v) chicken feather was purified and characterized. Keratinolytic enzyme secretion was the maximal at day 16 of the incubation period at pH 8 and $28^{\circ}C$. No relationship was detected between enzyme yield and increase of fungal biomass. The fraction obtained at 80% ammonium sulfate saturation showed 2.39-fold purification and was further purified by gel filtration in Sephadex G-100 followed by ion exchange chromatography on DEAE-Sephadex A-50, yielding an active protein peak showing 11.53-fold purification. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and zymograms indicated that the purified keratinase is a monomeric enzyme with 31 kDa molecular weight. The extracellular keratinase of A. flavus was active in a board range of pH ($7{\sim}10$) and temperature ($30^{\circ}C{\sim}70^{\circ}C$) profiles with the optimal for keratinase activity at pH 8 and $45^{\circ}C$. The keratinase activity was totally inhibited by protease inhibitors such as phenylmethylsulfonyl fluoride (PMSF), iodoacetic acid, and ethylenediaminetetraacetate (EDTA) while no reduction of activity by the addition of dithiothreitol (DTT) was observed. N-terminal amino acid sequences were up to 80% homologous with the fungal subtilisins produced by Fusarium culmorum. Therefore, on the basis of these characteristics, the keratinase of A. flavus K-03 is determined to be subtilisins-like.

• Journal of Food Hygiene and Safety
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• v.16 no.3
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• pp.206-211
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• 2001

This experiment was conducted to investigate the effects of mixed culture with mycotoxigenic and non-mycotoxigenic fungi on mycotoxin production. For this work, Aspegillus flavus (aflatoxin producing strain), Aspegillus niger (non-mycotoxigenic strain) and Penicillium griseofulvum (patulin producing strain)were cultured in 5 ml SLS medium for 15 days under single or mixed culture. Aflatoxin was determined by direct competitive ELISA, whereas, patulin was measured by HPLC. The mycelial growth, pH and total acidity were also observed by general methods. The mycelial growth was slightly decreased in the mixed culture, meanwhile total acidity was increased and pH was shown lower than that in single culture. Aspergillus flavus produced 145 $\mu\textrm{g}$/ml of aflatoxin for 12 days single culture, but in mixed culture, aflatoxin was decreased to 93%, and was shown as 10.16$\mu\textrm{g}$/ml level. Patulin production in mixed culture was also decreased to 69.3% and was shown only 23.72$\mu\textrm{g}$/ml level as compared with in single culture.

• 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.

• Applied Biological Chemistry
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• v.29 no.3
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• pp.255-259
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• 1986

The aflatoxin production by Aspergillus ATCC 15517 decreased in the mixed culture with Aspergillus awamori, Aspergillus kawachii, Aspergillus niger, Aspergillus oryzae, or with Aspergillus shirousamii to 1.3%, 13.8%, 1.3%, 0.7%, or 38.5% of that of monoculture respectively. These koji molds degraded $75%{\sim}100%$ of added aflatoxin $B_1\;(791{\mu}g/50ml\;YES medium)$. A. awamori secreted during growth aflatoxin degrading factor(s) which was heat-labile. The degraded aflatoxin by the factor(s) showed no toxicity against Bacillus megaterium NRRL B-1368.