• Journal of Microbiology and Biotechnology
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• v.28 no.12
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• pp.1971-1981
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• 2018

In this study, we investigated the altered enzymatic activities and metabolite profiles of koji fermented using varying permutations of Aspergillus oryzae and/or Bacillus amyloliquefaciens. Notably, the protease and ${\beta}$-glucosidase activities were manifold increased in co-inoculated (CO) koji samples (co-inoculation of A. oryzae and B. amyloliquefaciens). Furthermore, gas chromatography-mass spectrometry (GC-MS)-based metabolite profiling indicates that levels of amino acids, organic acids, sugars, sugar alcohols, fatty acids, nucleosides, and vitamins were distinctly higher in CO, SA (sequential inoculation of A. oryzae, followed by B. amyloliquefaciens), and SB (sequential inoculation of B. amyloliquefaciens, followed by A. oryzae). The multivariate principal component analysis (PCA) plot based on GC-MS datasets indicated a clustered pattern for MA and MB (koji samples inoculated either with A. oryzae or B. amyloliquefaciens) across PC2 (20.0%). In contrast, the CO, SA, and SB metabolite profiles displayed segregated patterns across PLS1 (22.2%) and PLS2 (21.1%) in the partial least-square discriminant analysis (PLS-DA) model. Intriguingly, the observed disparity in the levels of primary metabolites was engendered largely by higher relative levels of sugars and sugar alcohols in MA, SA, and CO koji samples, which was commensurate with the relative amylase activities in respective samples. Collectively, the present study emphasizes the utility of integrated biochemical and metabolomic approaches for achieving the optimal permutation of fermentative inocula for industrial koji preparation.

• Journal of Microbiology
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• v.43 no.6
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• pp.475-486
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• 2005

Fungal secondary metabolites constitute a wide variety of compounds which either playa vital role in agricultural, pharmaceutical and industrial contexts, or have devastating effects on agriculture, animal and human affairs by virtue of their toxigenicity. Owing to their beneficial and deleterious characteristics, these complex compounds and the genes responsible for their synthesis have been the subjects of extensive investigation by microbiologists and pharmacologists. A majority of the fungal secondary metabolic genes are classified as type I polyketide synthases (PKS) which are often clustered with other secondary metabolism related genes. In this review we discuss on the significance of our recent discovery of chalcone synthase (CHS) genes belonging to the type III PKS superfamily in an industrially important fungus, Aspergillus oryzae. CHS genes are known to playa vital role in the biosynthesis of flavonoids in plants. A comparative genome analyses revealed the unique character of A. oryzae with four CHS-like genes (csyA, csyB, csyC and csyD) amongst other Aspergilli (Aspergillus nidulans and Aspergillus fumigatus) which contained none of the CHS-like genes. Some other fungi such as Neurospora crassa, Fusarium graminearum, Magnaporthe grisea, Podospora anserina and Phanerochaete chrysosporium also contained putative type III PKSs, with a phylogenic distinction from bacteria and plants. The enzymatically active nature of these newly discovered homologues is expected owing to the conservation in the catalytic residues across the different species of plants and fungi, and also by the fact that a majority of these genes (csyA, csyB and csyD) were expressed in A. oryzae. While this finding brings filamentous fungi closer to plants and bacteria which until recently were the only ones considered to possess the type III PKSs, the presence of putative genes encoding other principal enzymes involved in the phenylpropanoid and flavonoid biosynthesis (viz., phenylalanine ammonia-lyase, cinnamic acid hydroxylase and p-coumarate CoA ligase) in the A. oryzae genome undoubtedly prove the extent of its metabolic diversity. Since many of these genes have not been identified earlier, knowledge on their corresponding products or activities remain undeciphered. In future, it is anticipated that these enzymes may be reasonable targets for metabolic engineering in fungi to produce agriculturally and nutritionally important metabolites.

• Fisheries and Aquatic Sciences
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• v.13 no.1
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• pp.12-17
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• 2010

This study was conducted to evaluate the potential of a microbial fermentation procedure to improve the functional qualities of seaweeds. Aspergillus oryzae, which has been used in traditional Korean fermented foods, was inoculated and cultivated in an aqueous extract of sea tangle (Laminaria japonica). Fermentation of the sea tangle extract by A. oryzae for 4 days resulted in a 3-fold increase in $\gamma$-aminobutyric acid (GABA) content. GABA is known to be a bioactive compound. Fungal fermentation of the extract also enhanced its antioxidant activity and increased its total content of phenolic compounds. It was assumed that these changes stemmed from the biodegradation of active compounds of the sea tangle packaged within its rigid structural matrix or occurred as result of fungal fermentation. These results suggested that the application of microbial fermentation to the processing of seaweeds will help in the development of processed foods to meet consumer demands.

• The Korea Journal of Herbology
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• v.26 no.4
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• pp.109-113
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• 2011

Objectives : We investigated the antioxidant and Antidiabetic effects of Cnidii Rhizoma Fermentata by Aspergillus oryzae and Aspergillus Kawauchi for 3days. Methods : In this study we compared Cnidii Rhizoma Fermentata with Aspergillus oryzae and Aspergillus Kawauchi that examined using reducing sugar, DPPH radical scavenging activity, ${\alpha}$-amylase and ${\alpha}$-glucosidase inhibitory activity. Also determined changes of pH and sugar content during fermentation for 3days. Results : The values for DPPH radical scavenging activity of Cnidii Rhizoma fermented by Aspergillus oryzae (86.6%) was higher than that of Aspergillus Kawauchi (77.9%). In ${\alpha}$-amylase inhibitory activity, fermented by Aspergillus Kawauchi had the highest inhibitory activity among other groups. But in ${\alpha}$-glucosidase inhibitory activity, fermented by Aspergillus oryzae had the highest inhibitory activity among other groups. While all groups of the sugar content increased During 3days fermentation, the pH was decreased. Conclusions : Based on these results, It was suggested that Cnidii Rhizoma Fermentata can be a useful and cost-effective resource for fuctional food and medicine.

• Journal of Applied Biological Chemistry
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• v.63 no.3
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• pp.229-234
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• 2020

Beer production with rice or other malt substitutes suffers from a lack of suitable enzymes for saccharification. For this reason, rice nuruk (fermentation starter) was tested as a starch replacement for malt in the saccharification process of beer production. The results of this study show that the enzyme activities of rice nuruk made with brewing fungi were higher than those of malt. Saccharification and glucoamylase activities were high in Aspergillus awamori KCCM 30790 and α-amylase activity was high in Aspergillus oryzae CF1003. Overall, malt beer had significantly higher alcohol, pH, total acid, volatile acids, amino acids, free amino nitrogen, bitterness unit and ΔE than rice nuruk beer. Where as Aspergillus awamori KCCM 30790 beer had significantly higher soluble solids, reducing sugar than malt beer. According to a sensory evaluation, malt beer was better color, flavor and Aspergillus oryzae CF1003 beer was better taste, texture, overall acceptability than other beer. Therefore Aspergillus awamori KCCM 30790 beer was suitable considering enzyme activities (saccharification, glucoalmylase) and physicochemical characteristics (soluble solids, reducing sugar). And then Aspergillus oryzae CF1003 beer was suitable considering sensory evaluation (taste, texture, overall acceptability). Therefore rice nuruk like Aspergillus awamori KCCM 30790 and Aspergillus oryzae CF1003 were suitable as a substitute material that can replace for malt in beer proceccing.

• Microbiology and Biotechnology Letters
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• v.10 no.3
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• pp.227-232
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• 1982

The methods determining the growth rate of mold on beans were investigated in order to compare the growth of Aspergillus oryzae on lupinseed to that on soybean. The growth of A. oryzae on cooked whole or paste form of bean substrates was evaluated by the measurements of colony diameter and hyphae length of the mold. The mold showed characteristic lag times to form the colony on different types of substrate. The growth of colony diameter was coincided with the increase in $\alpha$-amino nitrogen content of the substrate when the moisture level of the substrates was similar each other. The colony diameter and the cultivation time after the lag period showed a straight line relationship, from which the growth rate was estimated. in general, lupinseed paste allowed faster growth of A. oryzae than soybean paste at the initial growth phase. The lag time to form the colony was 24.0 hrs on lupinseed paste and 44.4 hrs on soybean paste. The growth rate after colony formation was, however, 7.05 mm/day for lupinseed paste and 8.83mm/day for soybean paste, which indicated that the growth rate after the lag period was faster on soybean compared to lupinseed. The sporulation time of the mold was related to the lag time for the colony formation. The measurement of hyphae length on whole beans could be used as a simple and rapid method of estimating the growth property of mold on different substrates.11 showed that the growth of A. oryzae was partly hindered by the thick hull of the lupinseed.

• The Korean Journal of Food And Nutrition
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• v.6 no.2
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• pp.89-95
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• 1993

A Nuluk, a Korean traditional Koji for brewing, was made with wheat flour and Aspergillus oryzae L2 which had a good aroma and strong abilities In producing saccharogenic and dextrogenic enzymes. The cultural conditions for the production of saccharogenic and proteolytic enzymes were tested. The productivity of dextrogenic enzyme was improved when Nuluk was made with unsteamed wheat flour as compared with steamed one, but that of proteolytic enzyme was reduced. The addition of water containing 0.5% hydrochloric acid was unfavorable for the production of those two enzymes. The optimum ratio of water added to wheat flour for the production of those two enzymes was 28$^{\circ}C$ on the basis of wheat flour, The productivity of saccharogenic enzyme was enhanced when the Nuluk was molded after 20 hours of precultivation, but that of proteolytic enzyme was reduced as compared with no molding. The optimum temperatures for the production of saccharogenic enzyme and proteolytic enzyme were 36$^{\circ}C$ and 28$^{\circ}C$, respectively.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.4
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• pp.253-262
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• 2000

Aspergillus is considered to be an attractive host for heterologous protein production because of its safety and ability to secrete large amounts of proteins. In order to obtain high productivity, thus far promoters of amylases have been most widely used in A. oryzae. Recent progress in cloning and expression analysis, including EST sequencing, revealed that glycolytic genes represent some of those most strongly expressed in A. oryzae. Therefore, promoters of glycolytic genes could be important alternatives to promoters of amylases because lower amounts of proteases are produced in the presence of glucose. Several A. oryzae transcription factors responsible for the induction and/or maximum expression of many industrially important genes encoding amylases and proteases have been cloned and characterized. In addition to the transcriptional regulatory factors, the gene encoding the largest subunit of RNa polymerase II, constituting the basic transcription machinery, has also been cloned from A. oryzae. This recently acquired understanding of the details of transcriptional regulatory mechanisms and factors will facilitate engineering flexible controls for the expression of proteins important for the fermentation industries.

• The Korean Journal of Mycology
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• v.40 no.4
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• pp.187-190
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• 2012

Various koji were prepared by fungi isolated from traditional nuruk and their quality characteristics were investigated. Acidity and saccharification power of their koji were ranged in 5.0~6.8 and 128sp~241sp. Nine fungi which were showed good quality and sensory evaluation were identified by analysis of their nucleotide sequences with PCR-amplified 18S rDNA internal transcribed spacer-1(ITS-1) and ITS-4 genes. Among them, six strains were identified as Aspergillus oryzae and the other strains were identified as Mycocladus corymbiferus, Rhizo-pus oryzae, Lichtheimia corymbifera.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.6
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• pp.987-994
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• 2000

Aspergillus coreanus NR 15-1, Asp. oryzae NR 15-3 and Asp. oryzae NR 2-5 isolated from traditional Korean nuruk were screened as parental strains producing starch hydrolyzing enzymes. They were mutagenized by N-methyl -N'-nitro-N-nitrosoguanidine (NTG) and mutants were isolated for analysis of various amylase activities and the ability of acid production. Among them, the mutants harboring high saccharogenic activity, dextrinogenic activity, and the ability of acid production were selected. Fifteen, six, and five strains of mutants were isolated from Asp. coreanus NR 15-1, Asp. oryzae NR 2-5, and Asp. oryzae NR 15-3, respectively followed by NTG mutagenesis. Among these mutants, thirteen strains were identified as auxotrophic mutants. \ulcorner (Arg.￣) mutant from Asp. coreanus NR 15-1 showed high glucoamylase activity and total acid productivity. Z6 (Ade.￣) mutant from Asp. oryzae NR 2-5 showed the highest $\alpha$-amylase activity, therefore \ulcorner and Z6 mutant were selected.