• Journal of Microbiology and Biotechnology
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• v.24 no.4
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• pp.447-452
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• 2014

To isolate novel and useful microbial enzymes from uncultured gastrointestinal microorganisms, a fecal microbial metagenomic library of the pygmy loris was constructed. The library was screened for amylolytic activity, and 8 of 50,000 recombinant clones showed amylolytic activity. Subcloning and sequence analysis of a positive clone led to the identification a novel gene (amyPL) coding for ${\alpha}$-amylase. AmyPL was expressed in Escherichia coli BL21 (DE3) and the purified AmyPL was enzymatically characterized. This study is the first to report the molecular and biochemical characterization of a novel ${\alpha}$-amylase from a gastrointestinal metagenomic library.

• 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.

• The Korean Journal of Mycology
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• v.30 no.2
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• pp.147-151
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• 2002

Trichoderma spp. are the aggressive causal agents for green mold disease on oyster mushroom (Pleurotus spp.) cultivation. Antifungal bacteria (KATB 99121, KATB 99122 and KATB 99123 strains) were isolated from the compost for Pleurotus ostreatus. Among these bacterial strains, KATB 99121 strain showed an excellent inhibitory activity to the pathogens for green molds such as T. harzianum, T. viride and T. hamatum and an animal pathogen, Candida albicans, but did not affect on the culture of Pleurotus ostreatus (2209, Chunchu 2 and Wonhyung strains). KATB 99121 strain secreted amylolytic, proteolytic and cellulolytic exoenzymes. KATB 99122 and KATB 99123 strains excreted amylolytic, proteolytic, cellulolytic, lipolytic exoenzymes and showed ${\beta}$-glucosidase activity. Further studies will be conducted on the development of microbial fungicides using the antagonistic bacteria for the control of green mold disease on Pleurotus spp.

• Korean Journal of Microbiology
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• v.41 no.2
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• pp.146-151
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• 2005

To construct an amylolytic industrial strain of Saccharomyces cerevisiae, the glucoamylase cDNA gene (GAl) from Aspergillus awamori was expressed under the control of the alcohol dehydrogenase gene promoter (ADC1p) and integrated into the chromosomes of industrial S. cerevisiae. An integrative cassette lacking bacterial ampicillin resistance gene but containing the GA1 gene, $\delta$ sequences of Ty1 retrotransposon as target sites for homologous recombination and S. cerevisiae aureobasidin A resistance gene (AUR1-C) as the selection marker was constructed to obtain a strain eligible for commercial use. Industrial S. cerevisiae transformed with this 15-integrative cassette efficiently secreted glucoamylase into the medium and grew on starch as the sole carbon source. The transformants were mitotically stable for 100 generations in nonselective medium.

• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1539-1545
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• 2010

In beer, glutathione works as the main antioxidant compound, which also correlates with the stability of the beer flavor. In addition, high residual sugars in beer contribute to major nonvolatile components, which are reflected in a high caloric content. Therefore, in this study, the Saccharomyces cerevisiae GSH1 gene encoding glutamylcysteine synthetase and the Saccharomycopsis fibuligera ALP1 gene encoding ${\alpha}$-amylase were coexpressed in industrial brewing yeast strain Y31 targeting the ${\alpha}$-acetolactate synthase (AHAS) gene (ILV2) and alcohol dehydrogenase gene (ADH2), resulting in the new recombinant strain TY3. The glutathione content in the fermentation broth of TY3 increased to 43.83 mg/l as compared with 33.34 mg/l in the fermentation broth of Y31. The recombinant strain showed a high ${\alpha}$-amylase activity and utilized more than 46% of the starch as the sole carbon source after 5 days. European Brewery Convention tube fermentation tests comparing the fermentation broths of TY3 and Y31 showed that the flavor stability index for TY3 was 1.3-fold higher, whereas its residual sugar concentration was 76.8% lower. Owing to the interruption of the ILV2 gene and ADH2 gene, the contents of diacetyl and acetaldehyde as off-flavor compounds were reduced by 56.93% and 31.25%, respectively, when compared with the contents in the Y31 fermentation broth. In addition, since no drug-resistant genes were introduced to the new recombinant strain, it should be more suitable for use in the beer industry, owing to its better flavor stability and other beneficial characteristics.

• Journal of Animal Science and Technology
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• v.62 no.2
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• pp.208-217
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• 2020

Heat stress negatively affects cattle productivity by reducing feed intake. In the present study, we assessed if the rumen microbiome composition of Hanwoo steers was altered by exposure to heat stress. Rumen samples were collected from four Hanwoo steers that were individually housed in climate-controlled chambers with 60% humidity and environmental temperatures of: 1) 15℃ (0-day group), 2) 35℃ for 3 days (3-day group), and 3) 35℃ for 6 days (6-day group). The total community DNA of samples was extracted, and 997,843 bacterial and 1,508,770 archaeal sequences were analyzed using next-generation sequencing. Assessment of the relative abundances revealed 15 major phyla of which Bacteroidetes was found to be the most dominant. After 3 days of heat stress exposure there were no significant changes in the rumen microbiome composition, except for a decrease in the Planctomycetes. However, after 6 days of heat stress exposure, we found that the relative abundance of fibrolytic Ruminococcaceae had decreased while that of lactate-producing Lactobacillaceae and amylolytic Prevotella and Ruminobacter had increased. The normal rumen microbiome of Hanwoo cattle was shown to be disrupted after 6 days of heat stress, which led to the decrease in fibrolytic bacteria that are sensitive to low pH and the increase in both lactate-producing and amylolytic bacteria. We have demonstrated that the microbiome composition of the rumen is affected by acute heat stress. Our findings may contribute to the development of different feeding strategies to restore heat stress-induced disruption of the rumen microbiome.

• Korean Journal of Food Science and Technology
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• v.34 no.2
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• pp.269-273
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• 2002

The amlyolytic enzymes $({\alpha}-amlyase,\;{\beta}-amlyase,\;glucoamlyase)$ and protease activities were studied during Kimchi fermentation. The optimum pH of Kimchi was 4.1 within 2 days at $20^{\circ}C$. The optimum acidity calculated as lactic acid was 0.44% within 2 days at $20^{\circ}C$. On the first day of fermentation, ${\alpha}-amlyase$ activity was reduced from 0.49 unit/mg protein to 0.20 unit/mg protein but increased in the later stage of fermentation. In case of ${\beta}-amlyase,\;glucoamlyase$ and protease showed the highest activity of 505.73, 13.43 and 1.72 unit/mg protein at the 2nd day of fermentation at $20^{\circ}C$. In the sensory evaluation of Kimchi were estimated taste, color, texture and overall acceptability. Overall acceptability of kimchi showed the highest score value on the 2nd day of fermentation, respectively.

• Journal of Microbiology and Biotechnology
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• v.31 no.2
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• pp.272-279
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• 2021

Two genes encoding probable α-ʟ-arabinofuranosidase (E.C. 3.2.1.55) isozymes (ABFs) with 92.3% amino acid sequence identity, ABF51A and ABF51B, were found from chromosomes 3 and 5 of Saccharomycopsis fibuligera KJJ81, an amylolytic yeast isolated from Korean wheat-based nuruk, respectively. Each open reading frame consists of 1,551 nucleotides and encodes a protein of 517 amino acids with the molecular mass of approximately 59 kDa. These isozymes share approximately 49% amino acid sequence identity with eukaryotic ABFs from filamentous fungi. The corresponding genes were cloned, functionally expressed, and purified from Escherichia coli. SfABF51A and SfABF51B showed the highest activities on p-nitrophenyl arabinofuranoside at 40~45℃ and pH 7.0 in sodium phosphate buffer and at 50℃ and pH 6.0 in sodium acetate buffer, respectively. These exoacting enzymes belonging to the glycoside hydrolase (GH) family 51 could hydrolyze arabinoxylo-oligosaccharides (AXOS) and arabino-oligosaccharides (AOS) to produce only ʟ-arabinose, whereas they could hardly degrade any polymeric substrates including arabinans and arabinoxylans. The detailed product analyses revealed that both SfABF51 isozymes can catalyze the versatile hydrolysis of α-(1,2)- and α-(1,3)-ʟ-arabinofuranosidic linkages of AXOS, and α-(1,2)-, α-(1,3)-, and α-(1,5)-linkages of linear and branched AOS. On the contrary, they have much lower activity against the α-(1,2)- and α-(1,3)-double-substituted substrates than the single-substituted ones. These hydrolases could potentially play important roles in the degradation and utilization of hemicellulosic biomass by S. fibuligera.

• 한국해양학회지
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• v.24 no.3
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• pp.157-164
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• 1989

Quantitative species distribution and primary productivity of phytoplankton were studied monthly from August, 1987 to July, 1988 along with the quantitative distribution of total heterotrophic bacterioplankton and three groups of physiologically chracteristic bacterioplankton in the intertidal and subtidal waters off Kum River Estuary, Yellow Sea. A total of 121 phytoplankton taxa including 102 diatoms occurred, and cell concentration ranged from 15 to 5451 (cells/ml). The great spatio-temporal variations of the number of phytoplankton species and cell concentration well reflected the environmental differences between the intertidal and subtidal waters. Primary productivity (in Piopt, mgC/$m^3$/hr) ranged from 0.6 to 27.3. Just after the phytoplankton bloom (March) Piopt was very low in April at station 1, where amylolytic bacterioplankton also showed quite low population density. The peaks of primary productivity were not always coincided with those of phytoplankton standing crop. The ratio of Piopt's between samples well indicated the environmental differences between the intertidal and subtidal waters. Little characteristic trend was found in the scatter diagrams of phytoplankton standing crop along the population densities of total heterotrophic bacterioplankton and the three groups of physiologically characteristic bacterioplankton. In summer the phytoplankton standing crop was minimum in contrast with the high population density of bacterioplankton, which implies the influx of much allochthonous orgainc matter from Kum River. The scatter diagrams of Piopt along bacterioplankton population density revealed some phenomena there. Piopt had highly positive correlation with the population density of amylolytie bacterioplankton($R^2$=0.84) and that of lipolytic bacterioplankton($R^2$=0.70) while total heterotrophic bacterioplankton and proteolytic bacterioplankton had lesser correlations with Piopt. From the regression lines the increase of unit Piopt (mgC/$m^3$/hr) in the study area was calculated to mean the increase of $9.0{\times}10$ cells/ml and $8.0{\times}10$ cells/ml of amylolytic bacterioplankton and lipolytic bacterioplankton, respectively.

• Korean Journal of Microbiology
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• v.23 no.3
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• pp.215-222
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• 1985

Distribution pattern and activity of heterotrophec bacteria were measured in the mudflat of Nakdong river estuary. In March and June, 1985, community sizes of amylolytic, lipolytic and proteolytic bacteria as well as total viable counts were measured. Vertical distribution of bacterial community size increased a few orders of magnitude from Narch to June. Heterotrophic activity was estimated in turnover time with $U-[^{14}C]-glucose$. Turnover time reduced considerably in June compared to that of March. To sxamine correlations for measured bacterial groups, turnover time and environmental factors, correlation coefficient matrix was obtained. These measured characteristics did not consistently correlate well with one another.