• Microbiology and Biotechnology Letters
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• v.15 no.6
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• pp.408-413
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• 1987

Microorganisms capable of degrading the raw naked barley were isolated from soil, and the amylase productivity of each strain was examined on plate contained 2％ raw naked barley. Of the fungi and actinomycetes tested, 71 strains were subjected to subsequent testing for amylase production, and 4 strains were selected as potent amylase producers. Among them, Strain No. 281 produced the most potent raw naked barley saccharifying enzyme, and was identified as genus Rhizopus from morphological and physiological studies. The ratio of raw starch saccharifying activity (RDA) of the crude enzyme derived from the Rhizopus sp. No. 281 was showed 2-3 fold higher than that of commercial enzyme when the raw naked barley was used as the substrate. In the case of uncooked alcohol fermentation using Rhizopus sp. No. 281 glucoamylase preparation, the alcohol yield of the broth was 2％ higher than that of the commercial enzyme.

• Korean Journal of Food Science and Technology
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• v.17 no.4
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• pp.258-264
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• 1985

The energy for cooking starch prior to saccharification could be saved by fermenting raw starch into ethanol using Aspergillus niger koji of wheat bran. Optimum cultivation time to produce glucoamylase was 4 days in wheat bran medium. The rate of saccharification from uncooked corn starch were optimum at pH 3.3 and 40-$50^{\circ}C$. Corn and sweet potato starch were saccharified more efficiently by wheat bran koji than other tested starch sources. 5 days of fermentation were required for optimum yield of ethanol using a mixture of AspergiUus niger koji and dried yeast. Final ethanol yields from raw corn, sweet potato, and rice starch with agitation at the rate of 100 rpm were about 95% at $30^{\circ}C$.

• Microbiology and Biotechnology Letters
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• v.26 no.5
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• pp.456-464
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• 1998

For the standardization and quality improvement of traditional Korean Nuruk, 10 strains of fungi, which were isolated from Nuruks and showed good productivity of the saccharogenic and dextrinogenic enzymes, acid and flavor, were selected and their enzymological characteristics and identification were carried out. Aspergillus spp. and Rhizopus sp. showed a high liquefying activity without regard to cultivation time, whereas the majority of strains except for Rhizopus sp. had decreasing saccharifying activity in proportion to the increase in cultivation time. Aspergillus spp. No.17-2, No.17-6 and Rhizopus sp. No.18-1 showed high liquefying and saccharifying activity after 15 and 30 day cultivation. The optimum temperature of most of these saccharogenic and dextrinogenic enzymes was from 40$^{\circ}C$ to 60$^{\circ}C$, and their optimum pH was extensive between pH 3 and pH 11. But Penicillium spp.(2 strains) and Rhizopus sp. showed low activity under the alkalic and acidic conditions. Among these isolated strains, 5 strains which had shown the high productivity of materials were identified as Aspergillus oryzae NR3-6 and Aspergillus oryzae NR17-6, Aspergillus penicilloides NR12-1, Penicillium expansum NR7-7 and Rhizopus oryzee NRl8-1, respectively. Five kinds of mixed culture were carried out and all of them showed a better productivity of saccharogenic and dextrinogenic enzymes than single culture. These results indicate that it is possible to make traditional Korean liquors of good quality by using these fungi.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.3
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• pp.576-581
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• 2004

Foxtail Millet Yakju were treated with heat ($65^{\circ}C$/15 min) or high hydrostatic pressure ($25^{\circ}C$ (RT) and $65^{\circ}C$ (HT)/400 MPa/10 min), and stored for 64 days at l$0^{\circ}C$, $25^{\circ}C$ and 37$^{\circ}C$. Changes in microbial counts, enzyme activity and quality of Yakju during storage were measured. Total viable cells were about 10$^2$ CFU/mL, and remained almost constant during storage at l$0^{\circ}C$ and $25^{\circ}C$, while decreased significantly at 37$^{\circ}C$, and undetected after 55 days of storage in heat- and pressure(RT)-treated, but after 25 days in pressure(HT)-treated Yakju. Lactic acid bacteria and yeast in heat- and pressure-treated Yakju were not detected during storage. The relative activities of a -amylase in heat- and pressure(RT)-treated were more than 100%, while those in pressure(HT)-treated were less than 40% during storage of 64 days at l$0^{\circ}C$. However, at $25^{\circ}C$ and 37$^{\circ}C$ the relative activities in untreated and pressure(RT)-treated were decreased greatly and then reached at the point of the activities of heat- and pressure(HT)-treated. The relative activities of glucoamylase in untreated and pressure(RT)-treated were decreased as the increase of the storage temperature during storage, while those in heat- and pressure(HT)-treated increased slightly as the increase of storage period at 1$0^{\circ}C$ and $25^{\circ}C$, and had no change at 37$^{\circ}C$. pH in heat- and pressure-treated had almost no change. Turbidity and reducing sugar in heat- and pressure-treated increased as the increase of storage temperature during storage.

• KSBB Journal
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• v.10 no.3
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• pp.304-316
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• 1995

For fuel alcohol production, enzymatic liquefaction and saccharification of tapioca starch by ${\alpha}$-amylase and glucoamylase were studied. The thermophilic ${\alpha}$-amylase Termamyl produced from Bacillus licheniformis gave a better liquefaction than the relalively low temperature enzyme BAN from B. subtilis. Oplimal temperature and pH with Termamyl were $90∼95^{\circ}C$ and 5.8, respectively. Minimal amount of Termamyl 240uc for a satisfactory liquefaction for a two-hour reaction was about 0.0125% (v/w) with respect to the mass of tapioca used. For saccharification experiments two enzymes, Novo AMG and Do-I1 enzymes were compared. The enzymatic activity of each enzyme was a little different depending on the substrate used and the latter was found to have a significant amount of ${\alpha}$-amylase activity. With Novo AMG optimal temperature was about $58^{\circ}C$ The pH optimum was 4.3 with maltose, however, with tapioca, no difference was observed between pH 4.3 and 5.7 which is a natural, unadjusted pH of liquefied tapioca. For 85% of completion of saccharification, it was necessary to use 0.0625% (v/w) of Novo AMG 400L for tapioca and to run the reaction for more than 10 hr, Packed volume of solid particles in tapioca slurry remained at around 30% during liquefaction and saccharification. This indicates that the removal of the solid particle before fermentation is not economically feasible at all, even though the solid particles make it very difficult to operate the bioreactor in a continuous mode with cell-recycle.

• Korean Journal of Food Science and Technology
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• v.29 no.6
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• pp.1158-1165
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• 1997

This study was designed to investigate the changes in microflora and enzyme activities of traditional kochujang during aging, which was prepared with a meju fermented for various periods of time. The number of bacteria was highest in all samples after 30 days of fermentation at $20^{\circ}C$, while that of mold in kochujang prepared with a 40-day- or 60-day-fermented meju was highest at 15th day of aging when yeast appeared. The activities of carbohydrases and proteases were highest in kochujang prepared with a 40-day-fermented meju followed by a 60-day-fermented meju. These results suggest that kochujang prepared with a meju aged for 40 days had the highest quality in terms of physicochemical, microbiological and enzyme activities of kochujang during fermentation.

• Korean Journal of Microbiology
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• v.20 no.3
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• pp.125-133
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• 1982

Mutational experiments were performed to imporve the cellulase productivity of Aspergillus phoenicis KU175, isolated from the southern part of Korea, as a high cellulase producer. By treatment ultra-violet light nad 4-NQO(4-Nitroquinoline-N-Oxide), mutation waas induced, and treatment ultra-violet light and 4-NQO (4-Nitroquinoline-N-Oxide), mutation was induced, and A.phoenicis KU175-115 was finally selected for its highest avicelase production. Avicelase production of the mutant was increased about 2 times compared with those of the wild strain. However, activities of other hydrolytic enzymes, such as amylase, protease and nuclease, of the mutant strain didn't show a marked difference compared with those of the nuclease, of the mutant strain didn't show a marked difference compared with the wild strain, except slight increase in ribonuclease activity and slight decrease in glucoamylase activity. Avicelases from the mutant strain selected were purified from wheat bran culture by successive salting out, followed by dialysis and column chromatography, and their charcteristics were compared with thosw of the wild strain. Avicelase was separated into three peaks in the mutant strain as well as in the case of wild strain. Avicelase II activity of the mutant strain was prominently higher than that of the wild strain, while avicelase I and III activities of those were equivalent. The optimal pH ranges and stability of avicelase II from the mutant strain were pH4-5 and pH3.5-6.0, respectively, as well as in the case of the wild strain. The optimal temperature and thermal stability of avicelase II from the mutant strain were $40{\sim}50^{\circ}C\;and\;20{\sim}55^{\circ}C$, respectively. These results were same as those of the wild strain. By the using of Eadie-Hofastee plot, $K_m\;and\;V_{max}$ of avicelase II from the mutant and the wild strain were calculated to be 2.29mg/ml and $4.84{\mu}g$ reducing sugar as glucose per min equally, from the line fitted to the data by the least square method. Activity of avicelase II from the mutant strain was slightly activated by $Mg^{++}\;but\;inhibited\;by\;Cu^{++}, \;Mn^{++}\;and\;Zn^{++}$, as well as in the case of the wild strain. Therefore, it was concluded that the mutant didn't induce the formation of another avicelase isozyme, or the changes in the properties of avicelase, but induce the changes in the productively of the same avicelase II by the action of regulatory gane.

• The Korean Journal of Mycology
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• v.13 no.4
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• pp.203-212
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• 1985

Of 450 strains isolated from the soil microbes collected in various locations in Korea, a strain had a strong inhibitory activity against bacterial ${\alpha}-amylase$ and was named strain DMC-72 of the genus Streptomyces. The amylase inhibitory metabolite produced by this strain was purified by means of acetone precipitation, adsorption on Amberlite IRC-50 and SP-Sephadex C-25. The inhibitor was found to be a derivative of oligosaccharides by spectral and chemical data. The inhibitor was stable at the pH range of $1{\sim}13$ and at $100^{\circ}C$ for half an hour, also inhibited other amylases such as salivary ${\alpha}-amylase$, pancreatic ${\alpha}-amylase$, fungal ${\alpha}-amylase$ and glucoamylase. However, it showed no inhibitory activity against ${\alpha}-glucosidase$, ${\beta}-glucosidase$, dextranase, and ${\beta}-amylase$. The kinetic studies of the inhibitor showed that its inhibitory effects on starch hydrolysis by ${\alpha}-amylase$ were noncompetitive.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.1
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• pp.52-59
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• 2005

The study was targeted to saccharify foodwastes with the cellulolytic and amylolytic enzymes obtained from culture supernatant of Trichoderma harzianum FJ1 and analyze the kinetics of the saccharification in order to enlarge the utilization in industrial application. T. harzianum FJ1 highly produced various cellulolytic (filter paperase 0.9, carboxymethyl cellulase 22.0, ${\beta}$-glucosidase 1.2, Avicelase 0.4, xylanase 30.8, as U/mL-supernatant) and amylolytic (${alpha}$-amylase 5.6, ${\beta}$-amylase 3.1, glucoamylase 2.6, as U/mL-supernatant) enzymes. The $23{\sim}98\;g/L$ of reducing sugars were obtained under various experimental conditions by changing FPase to between $0.2{\sim}0.6\;U/mL$ and foodwastes between $5{\sim}20\%$ (w/v), with fixed conditions at $50^{\circ}C$, pH 5.0, and 100 rpm for 24 h. As the enzymatic hydrolysis of foodwastes were performed in a heterogeneous solid-liquid reaction system, it was significantly influenced by enzyme and substrate concentrations used, where the pH and temperature were fixed at their experimental optima of 5.0 and $50^{\circ}C$, respectively. An empirical model was employed to simplify the kinetics of the saccharification reaction. The reducing sugars concentration (X, g/L) in the saccharification reaction was expressed by a power curve ($X=K{\cdot}t^n$) for the reaction time (t), where the coefficient, K and n. were related to functions of the enzymes concentrations (E) and foodwastes concentrations (S), as follow: $K=10.894{\cdot}Ln(E{\cdot}S^2)-56.768,\;n=0.0608{\cdot}(E/S)^{-0.2130}$. The kinetic developed to analyze the effective saccharification of foodwastes composed of complex organic compounds could adequately explain the cases under various saccharification conditions. The kinetics results would be available for reducing sugars production processes, with the reducing sugars obtained at a lower cost can be used as carbon and energy sources in various fermentation industries.

• 한국균학회소식:학술대회논문집
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• 2015.11a
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• pp.11-11
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• 2015

A total of sixty-six samples of Nuruk, a fermention starter used to make the Korean traditional rice wine, Makgeolli, were collected from central and southern regions of Korea in 2013 and 2014. We classified two groups of the Nuruk samples, "commercial" and "home-made", according to the manufacturing procedure and purpose of use. Commercial Nuruks were made in a controlled environment where the temperature and humidity are fixed and the final product is supplied to Makgeolli manufacturers. Home-made Nuruks were made under uncontrolled conditions in the naturally opened environment and were intended for use in the production of small amounts of home-brewed Makgeolli. We obtained more than five hundred isolates including filamentous fungi and yeasts from the Nuruk samples followed by identification of fungal species. Also we stored glycerol stocks of each single isolate at $-70^{\circ}C$. We identified the species of each isolate based on the sequences of ITS regions amplified with two different universal primer pairs. We also performed morphological characterization of the filamentous fungi and yeast species through observations under the microscope. We investigated the major fungal species of commercial and home-made Nuruks by counting the colony forming units (CFU) and analyzing the occurrence tendency of fungal species. While commercial Nuruks contained mostly high CFU of yeasts, home-made Nuruks showed relatively high occurrence of filamentous fungi. One of the representative Nuruk manufacturers used both domestic wheat bran and imported ones, mainly from US, as raw material. Depending on the source of ingredient, the fungal diversity was somewhat different. Another commercial Nuruk sample was collected twice, once in 2013 and again in 2014, and showed different diversity of fungal species in each year. Nuruks obtained from the southern regions of Korea and Jeju island showed high frequency of yeast such as Saccharomycopsis fibuligera and Pichia species as well as unique filamentous fungus, Monascus species. S. fibuligera was easily found in many Nuruk samples with high CFU. The major filamentous fungi were Aspergillus, Lichtheimia, Mucor and Penicillium species. In order to further our understanding of the isolates and their potential industrial applications, we assayed three enzymes, alpha amylase, glucoamylase and acid protease from 140 isolates out of about five hundred isolates and selected about 10 excellent strains with high enzyme activities. With these fungal isolates, we will perform omics analyses including genomics, transcriptomics, metabolic pathway analyses, and metabolomics followed by whole genome sequencing of unique isolates associated with the basic research of Nuruk and that also has applications in the Makgeolli making process.