• Microbiology and Biotechnology Letters
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• v.26 no.6
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• pp.490-496
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• 1998

Aspergillus oryzae producing high proteolytic enzyme was isolated from soybean koji and named tentatively A. oryzae O-1. A. oryzae U-1 was obtained by mutation of A. oryzae O-1 with ultraviolet (UV)-irradiation and produced 14 times higher pretense activity compared with A. oryzae O-1. A. oryzae E-1 was acquired by treatment of A. oryzae U-1 with 0.5 M ethylmethanesulfonate (EMS) for 6 min at 3$0^{\circ}C$ and produced 39 times higher proteolytic activity than A. oryzae O-1. With protoplast fusion between A. oryzae O-1 and A. oryzee E-1 in the presence of polyethylenegylcol (PEG)-CaCl$_2$, proteolytic activity was increased to 82 times compared to A. oryzae O-1, and the fusant was named A. oryzae PF. The activities of the cultures containing proteolytic enzymes produced by the strains were determined to be 0.23 U/$m\ell$ for A. oryzae O-1, 3.29 U/$m\ell$ for A. oryzae U-1, 8.91 U/$m\ell$ for A. oryzae E-1, and 19.0 U/$m\ell$ for A. oryzae PF.

• 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.

• Korean Journal of Food Science and Technology
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• v.34 no.5
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• pp.873-878
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• 2002

In order to industrialize traditional Meju-tasting Koji, the characteristics of Koji manufactured with bacteria and fungi found in traditional Meju were investigated. Bacillus subtilis B-4 and Aspergillus oryzae F-5 showing high enzyme activities including those of amylase and protease were used. L-value of Koji manufactured with B. subtilis B-4 had darker color and higher enzyme production than A. oryzae F-5 made one. B. subtilis B-4 made Koji showed higher enzyme production and sensony evaluation score than A. oryzae F-5 Koji. A. oryzae F-5 Koji showed superior color to B. subtilis B-4 Koji. Activity in color, capacity of enzyme production, viable cell count, and sensory evaluation of water activity controlled Koji was superior to the uncontrolled one.

• The Plant Pathology Journal
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• v.20 no.3
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• pp.229-233
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• 2004

The bacterial leaf blight, which is caused by Xantho-monas oryzae pv. oryzae, is the most damaging and intractable disease of rice. To identify the genes involved in the virulence mechanism of transposon TnS complex, which possesses a linearized transposon and transposase, was successfully introduced into X. oryzae pv. oryzae by electroporation. The transposon mutants were selected and confirm the presence of transposition in X. oryzae pv. oryzae by the PCR amplification of transposon fragments and the Southern hybridization using these mutants. Furthermore, transposon insertion sites in the mutant bacterial chromosome were deter-mined by direct genomic DNA sequencing using transposon-specific primers with ABI 3100 Genetic Analyzer. Efficiency of transposition was influenced mostly by the competence status of X. oryzae pv. oryzae cells and the conditions of electroporation. These results indicated that the insertion mutagenesis strategy could be applied to define function of uncharacterized genes in X. oryzae pv. oryzae.

• Microbiology and Biotechnology Letters
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• v.32 no.3
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• pp.256-264
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• 2004

A new primer set was developed for the detection and identification of Xanthomonas oryzae pv. oryzae, the bacterial leaf blight (BLB) pathogen in rice plant. The nucleotide sequence of hpaA gene was determined from X. o. pv. oryzae str. KACC10331, and the sequence information was used to design primers for the application of the polymerase chain reaction (PCR). The nucleotide sequence of hpaA from X. o. pv. oryzae str. KACC 10331 was aligned with those of X. campestris pv. vesicatoria, X. campestris pv. campestris, X. axonopodis pv. citri, and X. axonopodis pv. glycines. Based on these results, a primer set(XOF and XOR) was designed for the specific detection of hpaA in X. o. pv. oryzae. The length of PCR products amplified using the primer set was 534-bp. The PCR product was detected from only X. o. pv. oryzae among other Xanthomonas strains and reference bacteria. This product was used to confirm the conservation of hpaA among Xanthomonas strains by Southern-blotting. Furthermore, PCR amplification with XOF and XOR was used to detect the pathogen in an artificially infected leaf. The sensitivity of PCR detection in the pure culture suspension was also determined. This PCR-based detection methods will be a useful method for the detection and identification of X. o. pv. oryzae as well as disease forecasting.

• 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.

• 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.

• Microbiology and Biotechnology Letters
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• v.30 no.1
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• pp.46-50
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• 2002

To investigate the quality of Koji produced by Cordyceps sp., a mixed culture of Aspergillus oryzae and the Cordyceps sp. were inoculated in Koji making. Viable cell counts and counts of fungi spore during Koji making for 120 hours were increased rapidly within 24-48 hours, after then their changes were not shown significantly during Koji making. The activities of amylase and protease in Koji inoculated with the mixed culture of A. oryzae and Cordyceps sp. were superior to one with A. oryzae only. The sensory evaluation on the Koji with 0.5％ (w/w) A. oryzae and 0.5％ (w/w) Cordyceps sp. were evaluated to be superior to ones. The best quality of Koji is prepared from the mixture of Cordyceps sp. and A. oryzae with the ratio of 0.5％ and 0.5％ (w/w).

• The Korean Journal of Food And Nutrition
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• v.9 no.4
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• pp.398-403
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• 1996

To produce low salt fermented anchovy by an accelerated method with Asp. oryzae and Bacillus sp. koji and taste properties after the 60 day fermentation were examined. The main free amino acids of 60 day fermented anchovy paste were valine, isoleucine, proline, alanine, lysine, glutamic acid and aspartic acid. Total amount of free amino acids was the highest in non koji anchovy paste wit 2,624.76mg%. Among the koji added samples, Asp. oryzae koji added on was the highest in the amount o free amino acids. Hypoxanthine accounted for 84.14~95.4% of total nucleotides and their related compounds; Asp. oryzae koji added anchovy paste was the highest in nucleotides other related. Citric acid and lactic acid accounted for 94.9~96.7% of total non-volatile organic acids; Asp. oryzae koji added sample was the highest in non-volatile organic acids with 287.93mg%. The Hunter a and b values gradually increased during the fermentation, but the L value decreased until day 30 or 40 and increased steadily after that. The a and b values were higher in the use of Asp. oryzae koji than in Bccillus sp. koji, but the L value was to the contrary. The Asp. oryzae koji added anchovy paste was good in the aspect of color and taste compared to others. In the aspect of odor, the anchovy paste using the mixture of Asp. oryzae and Bacillus sp. koji was the best. Overall aceptability of sensory evaluation was higher in the mixture of Asp. oryzae and Bacillus sp. koji tan in the others.

• Mycobiology
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• v.43 no.4
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• pp.415-422
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• 2015

The study aimed to evaluate the probiotic properties, antioxidant activity and fermentative capacity of Acremonium charticola and Rhizopus oryzae isolated from the Indonesian fermented dried cassava, with particular application on poultry. A. charticola inhibited the growth of Escherichia coli and Aspergillus flavus. A. charticola and R. oryzae grew in potato dextrose agar (PDA) adjusted to pH 3 and 8 or in PDA supplemented with bile salt up to 0.8%. After soaking for 8 hr, the survival rate of A. charticola in the simulated gastric juice (pH 2) and bile solutions (2% bile salt) was lower than that of R. oryzae. A. charticola and R. oryzae exhibited strong antioxidant activities. Compared to unfermented cassava pulp (control), the fibre content of cassava pulp tended to be lower after fermentation with A. charticola for 14 days. The populations of A. charticola and R. oryzae were significantly higher in fermented cassava pulp than in unfermented one. Coliform was higher in cassava pulp fermented with R. oryzae or A. charticola + R. oryzae compared to control after 7 days of fermentation, however, the bacteria were not different between A. charticola-fermented cassava pulp and control. Lactic acid bacteria (LAB) were higher in A. charticola- and R. oryzae-fermented cassava pulp than those in control, however, no difference of LAB was observed between A. charticola + R. oryzae-fermented cassava pulp and control. In conclusion, A. charticola exhibited antibacterial, antifungal and antioxidant activity, gastrointestinal persistence and fermentative capacity that may be beneficial for poultry industry.