• Korean Journal of Microbiology
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• v.27 no.3
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• pp.216-220
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• 1989

As the bsic study about protoplast fusion of amylolytic fungus Aspergillus oryze and nonamyloytic sugar fermenter, Saccaromyces cerevsisae, the intraspecific protoplast fusion of A. oryzae was carried out and the properties of the obtained fusants were investigated. For protoplast fomation from mycellia of auxotrophs, Novozyme 234 as lytic enzyme was the most effective and optimal pH was determined to be pH 5.5-6.0. When the two types of protoplasts were treated with a fusogen including 30% PEG4000, they fused effectively and most of fusants were heterokaryons. Protoplasts aggregated with 30% PEG4000 after fusion treatment were observed by the microscope. Protoplast regeneration frequency was 1.46 to 13.8% and complementation frequency of fusion was 0.12 to 0.16. Fusant strains had a 1.5-fold DNA content compared to that of parent strain. And amylase activity was intermediate between those of parent strains.

• The Korean Journal of Mycology
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• v.42 no.4
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• pp.282-288
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• 2014

In this study, we attempted to isolate fungi from soybean fermented foods produced in Sunchang County and to identify Aspergillus oryzae from fungal isolates. Ten fungal isolates were identified with ${\beta}$-tubulin gene. According to the sequences of ${\beta}$-tubulin gene, ten fungal isolates were identified as A. oryzae/flavus complex. For further identification of the ten of fungal isolates, omtA gene, one gene of the aflatoxin biosynthesis gene cluster, was sequenced and the sequences were compared with those of A. oryzae and A. flavus strains from the GenBank database. In addition, identification of the ten fungal isolates was further confirmed using the PCR amplicon of norB and cypA intergenic region, in which a deletion was recognized relative to A. flavus and A. parasiticus. The amplicon size of the ten fungal isolate strains was smaller than those of A. flavus and A. parasiticus, but the same as that of the reference A. oryzae strain. These results indicated that the ten isolates should be identified as A. oryzae. The protease activity in rice koji made with 6, 13, 17, 27, 37 and 38 of strain, respectively was twice higher than that in control. The kojis made with nine of the A. oryzae isolates, respectively, did not produce aflatoxin, suggesting that the strains could possibly be used as starters for soybean products.

• Korean Journal of Microbiology
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• v.31 no.1
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• pp.54-62
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• 1993

Intracellular adenosine deaminase (ADA) from Aspergillus oryzae was purified using ammonium sulfate fractionation, a DEAE-Sephadex A-50 anion exchange chromatography, an ultrafiltration using a PM 10 membrane and two times of Sephadex G-100 gel filtration chromatography. The enzyme was purified 151 fold with a 9% recovery. Purified enzyme gave a single protein band with a molecular weight of 105,000 delton. The enzyme was reasonably stable. The enzyme activity was kept even after 1 hr incubation at 55.deg.C, but decreased significantly at 60.deg.C. The pH optimum was found to be from 6.5 to 7.5. Among tested compounds, the substrate activity was found with adenosine, adenine arainofuranoside, formymcin A, 2'-deoxyadenosine, 3'-deoxyadenosine, 2', 3'-isopropylidene adenosine, 2,6-diaminopurine deoxyriboside, .betha.-nicotinamide adenine dinucleotide (reduced form), 6-chloropurine riboside, 2'-adenine monophosphate (AMP), 3'-AMP and 5'-AMP. The values of Km of adenosine and 2'-deoxyadenosine were calculated to be 500 and .$710\mu$m, respectively. ADA was sensitivite to $Zn^{2+}$, $^Cu{2+}$ and $Fe^{3+}$, p-chloromercuribenzoate and mersalyl acid inactivated the enzyme. The activity of enzyme was not changed when ADA was incubated with dithiothreititol, 2-mercaptoethanol, N-ethylmaleimide, iodoacetic acid and iodoacetamide.

• Korean Journal of Microbiology
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• v.27 no.3
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• pp.221-224
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• 1989

Amylolytic filamentous fungus, Aspergillus oryzae and nonamylolytic sugar fermentable yeast, Saccharomyces cerevisiae were fused by protoplast fusion in order to develope microorganisms having their intergrated function. Aminoacid auxotrophic properties were used as a genetic marker of protoplast fusion, and 35% PEG 4000 was used as a fusogenic agent. Complementation frequengy of fusion was $4.6\times 10^{-6}$ Obtained fusants showed the morphology of yeast strains, the amylase activity and the ethanol productivity. Among the properties of the fusants, morphology and prototrophic property were sustained stably but their ethanol productivity from starch was reduced. Although fusant strains had 0.5-fold ethanol productivity compared to that of S. cerevisiae in glucose medium, they produced ethanol from strach by direct fermentation.

• Journal of Life Science
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• v.16 no.1
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• pp.136-140
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• 2006

Ginseng was fermented by Aspergillus oryzae to search metabolites on the basis of increased biological activity and modified structure. From this research, two biotransformed compounds (WC2-2-1 and WC2-2-2) were detected and isolated through several chromatographic techniques. WC2-2-2 was confirmed to biologically active compound K by TLC, HPLC, and mass spectroscopy, while WC2-2-1 was going to be identified until now. In biological activity, both WG2-2-1 and WG2-2-2 exhibit the cytotoxicity on PC-3 cells, but WG2-2-2 was more active than WG2-2-1. It is supposed that WG2-2-1 is an intermediate metabolite transforming to final WG2-2-2, compound K.

• Korean Journal of Food Science and Technology
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• v.31 no.6
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• pp.1570-1576
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• 1999

Quality characteristics of 3 kinds of kochujang were investigated during 6 months of fermentation in order to improve the quality of industrial kochujang to that of traditional one. Three different kinds of kochujang were prepared using Aspergillus oryzae, Aspergillus oryzae plus Bacillus licheniformis and Aspergillus oryzae, Bacillus licheniformis plus Saccharomyces rouxii. Kochujang prepared with all three microorganisms contained lesser amounts of free sugar than the other preparations and glucose was the most abundant free sugar found. Ethanol was the most abundant amount among alcohols in kochujang. Kochujang prepared with A. oryzae, B. licheniformis plus S. rouxii has more amino nitrogen content than that prepared with A. oryzae only. Capsaicin contents of kochujang decreased slightly in all three kochujang, but there was no significant differences. The results of sensory evaluation showed statistically significant difference among three samples and panel members preferred the taste and flavor of kochujang prepared with all three microorganisms to those of the other samples.

• KSBB Journal
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• v.14 no.3
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• pp.259-263
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• 1999

Fish oil was hydrolyzed with Aspergillus oryzae lipase, Lipolase-100T. The hydrolysis characteristics of Lipolsae-100T were investigated. Lipolase-100T showed 1,3-positional specificity which hydrolyzed acyl chains combined on the 1 or 3 position of triglyceride into free fatty acids. Lipolase-100T represented another property that the saturated fatty acids composing the triglyceride were hydrolyzed more easily that the polyunsaturated fatty acids(PUFAs). n-3 PUFAs, such as C16:4, C20:5 and C22:6, were hardly hydrolyzed, so that the concentrations of those in the mixture of glycerides were increased according to hydrolysis time. Especially docosahexaenoic acid(DHA), C22:6 showed the highest increase in the concentration. This result explained that n-3 PUFAs were combined on 2-position of triglyceride. When the hydrolysis of fish oil with Lipolase-100T 0.4 wt% was performed for 120 hr, n-3 PUFAs wt% was increased to 50 wt% in the mixture of glycerides. This result was obtained due to the 1,3-positional specificity of Lipolase-100T and positional specificity of n-3 PUFAs.

• Journal of Physiology & Pathology in Korean Medicine
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• v.38 no.1
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• pp.16-21
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• 2024

Haepyoijin-tang and its main components have been used for phlegm, cough and dyspnea. Using a respiratory inflammation model, we intend to reveal the anti-inflammatory effect and pharmacological mechanism of Haepyoijin-tang. We induced the respiratory inflammation model by Aspergillus oryzae protease and ovalbumin administration. Female Balb/c mice (8 weeks old) were classified into four groups as follows: saline control group, aspergillus oryzae protease and ovalbumin induced respiratory inflammation group (vehicle), inflammation with Haepyoijin-tang (200 mg/kg) administration group, inflammation with dexamethasone (5 mg/kg) administration group (n=7). To identify the anti-inflammatory effects of Haepyoijin-tang water extracts, we measured the inflammatory cell number in bronchoalveolar lavage fluid (BALF) and total live lung cell number. In addition, we checked eosinophil ratio and number in BALF. And Interleukin (IL)-5 level was also measured in lung cell culture supernatant. To confirm the mechanism of anti-inflammatory effects, we analyzed the activated helper T cell (CD4⁺CD25⁺ cell) and Th2 cell (CD4⁺GATA3⁺ cell) ratio and number in lung by using flow cytometry. Finally, we attempted to confirm the immune mechanism by measuring the ratio and number of regulatory T cells (CD4⁺Foxp3⁺ cell). Haepyoijin-tang extracts treatment diminished inflammatory cell, especially, eosinophil number in BALF and total live lung cell number. Moreover, IL-5 level was reduced in Haepyoijin-tang treated group. Surprisingly, Haepyoijin-tang extracts administration not only decreased the activated helper T cell but also Th2 cell population in lung. Additionally, regulatory T cell population was increased in Haepyoijin-tang administration group. Our findings proved that Haepyoijin-tang extract have anti-inflammatory efficacy by suppressing Th2 cell activation and promoting regulatory T cell population.

• KSBB Journal
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• v.10 no.1
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• pp.38-45
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• 1995

The expression of Aspergillus nidulans glyceraldehyde-3-phosphate dehydrogenase (gpdA) and trpC promoters in A. oryzae were compared using E. coli lacZ gents fusions. The specific activities of the expressed E. coli $\beta$-galactosidase in A. oryzae transformants containing the A. nidulans gpdA promoter were around 2,000 units per ug of protein. The specific activities of transformants containing the A. nidulans trpC promoter were very low, ranging from 10.5 to 52.3 units per ug of protein. These results showed that the expression of the A. nidulans gpdA promoter in A. oryzae was approximately 70 times greater than the A. nidulans trpC promoter. In western blot analysis, immunoreactive bands of a imlilar molecular weight as the E. coli $\beta$-galactosidase were observed in A. oryzae carrying the gpdA-lacZ fusion and to a lesser intensity in those carrying the tvpC-lacZ fusion. Southern analysis showed that the higher expression of the gpdA-lacZ fusion as compared to the trpC-lacZ fusion was not due a greater number of integrated plasmids.

• Journal of Nutrition and Health
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• v.9 no.3
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• pp.18-25
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• 1976

To explain the change of the pungent principles of red pepper, capsaicin, during storage, the relation between infecting mold and capsaicin was studied. Results obtained were summarized as follows; 1. Superior strains which showed good growth and high decomposition activity in their culture broths, Aspergillus oryzae and Aspergillus sp. KF-7 were obtained from the contaminated red pepper fruits. 2. The weight of dry matter was increased at low concentration of capsaicin $(2{\sim}3{\mu}g/ml)$ but it was decreased at high concentration. 3. The residual capsaicin in the culture medium was reduced as half as such after 1 week, and almost disappeared after 2 weeks. At $30^{\circ}C$ and $pH\;7{\sim}9$ of culturing condition, the amounts of reducing capsaicin reached to the maximum. 4. At ground red pepper fruits as culture medium, Aspergillus sp. KF-7 could not glow at all, but Aspergillus oryzae showed good growth at 13% of moisture contents and the residual of capsaicin of red pepper fruits after 40days of cultivation time was 45%. 5. At ground red pepper fruits seed as culture medium, Aspergillus oryzae and Aspergillus sp. KF-7 showed good growth, and the residual of capsaicin after 4 weeks of cultivation time was 55% and 38% respectively.