• Journal of Microbiology and Biotechnology
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• v.22 no.7
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• pp.990-999
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• 2012

The microbial biosynthesis of fatty acid of lipid metabolism, which can be used as precursors for the production of fuels of chemicals from renewable carbon sources, has attracted significant attention in recent years. The regulation of fatty acid biosynthesis pathways has been mainly studied in a model prokaryote, Escherichia coli. During the recent period, global regulation of fatty acid metabolic pathways has been demonstrated in another model prokaryote, Bacillus subtilis, as well as in Streptococcus pneumonia. The goal of this study was to increase the production of long-chain fatty acids by developing recombinant E. coli strains that were improved by an elongation cycle of fatty acid synthesis (FAS). The fabB, fabG, fabZ, and fabI genes, all homologous of E. coli, were induced to improve the enzymatic activities for the purpose of overexpressing components of the elongation cycle in the FAS pathway through metabolic engineering. The ${\beta}$-oxoacyl-ACP synthase enzyme catalyzed the addition of acyl-ACP to malonyl-ACP to generate ${\beta}$-oxoacyl-ACP. The enzyme encoded by the fabG gene converted ${\beta}$-oxoacyl-ACP to ${\beta}$-hydroxyacyl-ACP, the fabZ catalyzed the dehydration of ${\beta}$-3-hydroxyacyl-ACP to trans-2-acyl-ACP, and the fabI gene converted trans-2-acyl-ACP to acyl-ACP for long-chain fatty acids. In vivo productivity of total lipids and fatty acids was analyzed to confirm the changes and effects of the inserted genes in E. coli. As a result, lipid was increased 2.16-fold higher and hexadecanoic acid was produced 2.77-fold higher in E. coli JES1030, one of the developed recombinants through this study, than those from the wild-type E. coli.

• Microbiology and Biotechnology Letters
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• v.48 no.2
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• pp.148-157
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• 2020

Eucalyptus oil is a rich source of bioactive compounds with a variety of biological activities and is widely used in traditional medicine. Eucalyptus citriodora is cultivated for the production of essential oils. However, the mode of antibacterial action of essential oils from E. citriodora is not well-known. This study aimed to determine the chemical components, microbial inhibitory effect, and mechanism of action of the essential oil from E. citriodora. The oil was extracted from E. citriodora leaves by hydro-distillation and the chemical components were analyzed using gas chromatography-mass spectrometry. The antibacterial activities of eucalyptus oil against gram-positive bacteria (Bacillus subtilis, Staphylococcus aureus, and Staphylococcus intermedius) and gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa) were screened by disc diffusion method and quantitative analysis was conducted by the microdilution method. The mechanism of action of the extracted essential oil was observed using SEM and analyzed by SDS-PAGE. The major components of E. citriodora oil were citronellal (60.55 ± 0.07%), followed by dl-isopulegol (10.57 ± 0.02%) and citronellol (9.04 ± 0.03%). The antibacterial screening indicated that E. citriodora oil exhibited prominent activity against all tested strains. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against B. subtilis were 0.5% and 1.0%, respectively. The MIC and MBC concentrations against S. aureus, S. intermedius, E. coli, and P. aeruginosa were 1% and 2%, respectively. As observed by SEM, the antibacterial mechanism of E. citriodora oil involved cell wall damage; SDS-PAGE revealed decrease in protein bands compared to untreated bacteria. Thus, E. citriodora oil showed significant antimicrobial properties and caused cellular damage.

• Computers and Concrete
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• v.27 no.4
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• pp.319-332
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• 2021

The performance of gene expression programming (GEP) in predicting the compressive strength of bacteria-incorporated geopolymer concrete (GPC) was examined in this study. Ground-granulated blast-furnace slag (GGBS), new bacterial strains, fly ash (FA), silica fume (SF), metakaolin (MK), and manufactured sand were used as ingredients in the concrete mixture. For the geopolymer preparation, an 8 M sodium hydroxide (NaOH) solution was used, and the ambient curing temperature (28℃) was maintained for all mixtures. The ratio of sodium silicate (Na2SiO3) to NaOH was 2.33, and the ratio of alkaline liquid to binder was 0.35. Based on experimental data collected from the literature, an evolutionary-based algorithm (GEP) was proposed to develop new predictive models for estimating the compressive strength of GPC containing bacteria. Data were classified into training and testing sets to obtain a closed-form solution using GEP. Independent variables for the model were the constituent materials of GPC, such as FA, MK, SF, and Bacillus bacteria. A total of six GEP formulations were developed for predicting the compressive strength of bacteria-incorporated GPC obtained at 1, 3, 7, 28, 56, and 90 days of curing. 80% and 20% of the data were used for training and testing the models, respectively. R2 values in the range of 0.9747 and 0.9950 (including train and test dataset) were obtained for the concrete samples, which showed that GEP can be used to predict the compressive strength of GPC containing bacteria with minimal error. Moreover, the GEP models were in good agreement with the experimental datasets and were robust and reliable. The models developed could serve as a tool for concrete constructors using geopolymers within the framework of this research.

• Journal of Microbiology and Biotechnology
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• v.32 no.8
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• pp.989-1002
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• 2022

Cephalopods, in particular octopus (Octopus vulgaris), have the ability to alter their appearance or body pattern by showing a wide range of camouflage by virtue of their chromatophores, which contain nanostructured granules of ommochrome pigments. Recently, the antioxidant and antimicrobial activities of ommochromes have become of great interest; therefore, in this study, the pH-dependent redox effect of the extraction solvent on the antioxidant potential and the structural characterization of the pigments were evaluated. Cell viability was determined by the microdilution method in broth by turbidity, MTT, resazurin, as well as fluorescence microscopy kit assays. A Live/Dead Double Staining Kit and an ROS Kit were used to elucidate the possible inhibitory mechanisms of ommochromes against bacterial and fungal strains. The results obtained revealed that the redox state alters the color changes of the ommochromes and is dependent on the pH in the extraction solvent. Natural phenoxazinone (ommochromes) is moderately toxic to the pathogens Staphylococcus aureus, Bacillus subtilis, Salmonella Typhimurium and Candida albicans, while the species Pseudomonas aeruginosa and Pseudomonas fluorescens, and the filamentous fungi Aspergillus parasiticus, Alternaria spp. and Fusarium verticillioides, were tolerant to these pigments. UV/visible spectral scanning and Fourier- transform infrared spectroscopy (FTIR) suggest the presence of reduced ommatin in methanol/ HCl extract with high intrinsic fluorescence.

• Journal of Animal Science and Technology
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• v.49 no.5
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• pp.667-676
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• 2007

This study was conducted to determine effects of cellulolytic microbes inoculation to sawdust-based spent mushroom substrate(SMS) during deepstacking on fermentation parameters, total microbial counts and cellulolytic enzyme activity and to on SMS nutrients utilization by sheep. For sheep metabolism trials, six sheep(ram, average 54.8kg) were fed a Control diet(70% concentrates, 15% rice straw and 15% SMS with no microbial treatment on a dry basis) and a Treatment diet(the same diet including SMS with a microbial treatment) for 2 trials. Spent mushroom substrates with or without a microbial(4 strains including 1 strain of Enterobacter ludwigii, 1 strain of Bacillus cereus and 2 strains of Bacillus subtillis) treatment (1% of SMS on wet basis) were deepstacked for 7 days. The internal temperatures in 1.2 M/T of SMS deepstacks reached to 50±5℃ within 7 days of storage. Total microbial counts remarkably decreased (P<0.05) with a deepstacking process and were not affected(P>0.05) by the microbial treatment. For fibrolytic enzyme activity, CMCase and xylanase activities were decreased(P<0.05) by a deepstacking process. After deepstacking, the microbial treatment showed about 2.5-times higher(P<0.05) for CMCase activity and about 4-times higher(P<0.05) for xylanase activity than those of the Control. Activities of ligninolytic enzymes such as laccase and MnP were not affected by the microbial treatment. The sheep fed the microbially treated SMS diet had a tendency of greater total tract digestibilities of ash(P=0.051), NFE (P=0.071), hemicellulose(P=0.087) and NDF(P=0.096) than those fed the untreated SMS diet. Nitrogen balance of sheep was not affected(P>0.05) by feeding of microbially treated SMS. Accordingly, these results indicate that cellulolytic microbes inoculation during deepstacking of SMS may improve the bio- utilization of SMS by sheep.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.6
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• pp.695-702
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• 2017

This study was carried out to investigate the effects of seed vinegar on antioxidant activity and antimicrobial activities of concentrated blueberry vinegar (CBV). Of the nine strains of yeast and six strains of acetic acid bacteria provided by the Microbial Institute for Fermentation Industry, each strain of yeast (Saccharomyces cerevisiae SRCM 100610, showing the highest ethanol content) and acetic acid bacteria (Acetobacter pasteurianus SRCM 101342, showing the highest total acidity) was selected for production of CBVs. Sugar content, pH, total acidity, total phenolic content (TPC), and browning intensity (280 nm and 420 nm) in CBVs using concentrated blueberry juice were $11.05{\sim}12.70^{\circ}Brix$, 2.63~2.98, 1.65~5.72%, 3.03~4.24 mg/mL, 0.95~1.50, and 0.11~0.20, respectively. Sugar content and total acidity of CBVs increased upon addition of seed vinegar, whereas pH, TPC, and browning intensity decreased. Of all CBVs with various additions of seed vinegar, the control showed the lowest $EC_{50}$ values in DPPH radical scavenging assay, ABTS radical scavenging assay, and reducing power (23.80, 19.48, and 79.21 dilution factor, respectively), whereas the 40% seed vinegar group showed the highest clear zone diameter values for Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, and Bacillus cereus (4.31, 4.59, 5.81, and 3.97, respectively). Antioxidant activities of CBVs were closely correlated with their TPC, browning intensity at 280 nm, pH, and total acidity values, showing correlation determination coefficient ($R^2$) values higher 0.82. However, antimicrobial activities of CBVs were closely correlated with their pH and total acidity values, showing higher $R^2$ values more than 0.92. These results suggest that CBVs using concentrated blueberry juice, S. cerevisiae SRCM 100610, and A. pasteurianus SRCM 101342 may be useful as potentially functional foods for enhancing health.

• Journal of Food Hygiene and Safety
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• v.29 no.3
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• pp.189-194
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• 2014

This study was conducted to evaluate microorganism contamination of food utensils and service facilities in school and to prevent hazards by food poisoning occurrence. As a result, the highest number of microorganism growth plate ($12.3{\pm}2.6$) was detected in total bacteria test plate, and also observed $10.3{\pm}3.9$ growth plates in Staphylococcus aureus test plate and $9.5{\pm}3.9$ growth plates in E. coli and coliform bacteria test plate. But we could detect to the lowest number of growth plates ($1.5{\pm}1.0$) in Vibrio test plate. We also assessed that floors were appeared to the highest microorganism contamination rate in food utensils and service facilities. Therefore, $4.5{\pm}0.6$ growth plates was detected in pre-operation floor and $4.3{\pm}1.0$ growth plates in floor. And high level of microorganism contamination also observed in tables as $3.3{\pm}1.0$ growth plates in cooking table and $3.0{\pm}0.0$ growth plates in dining table. The level of microorganism contamination of food utensils such as kitchen knife, cutting board, and food tray were lower than that in food service facilities. We analysed microorganism contamination according to purpose of use in kitchen knifes and cutting boards. The microorganism contamination rate in fish kitchen knife ($2.0{\pm}0.8$) and fish cutting board ($1.3{\pm}1.5$) were slightly higher than that of others purpose of use. As a result of microorganism identification, various strains of microorganism were contaminated in food service facilities and some strains could detected more than two times. Especially, Staphylococcus aureus was repeatedly identified in cooking table, trench, and kitchen knife. Bacillus cereus was identified in kitchen knife, and then Pseudomonas fluorescens and Pseudomonas aeruginosa were also detected in food utensils and service facilities as known to food spoilage microorganisms. Klebsiella pneumoniae was detected four times repeat, which widely distribute natural environment as normal bacterial flora but sometimes cause acute pneumonia. These results suggest that food utensils and service facilities are contaminated with not only major food poisoning microorganisms such as Staphylococcus aureus, but also food spoilage microorganisms. Taken together, strict personal hygiene control and efficient food service facilities management will be needed to enhance food safety in school feeding and to improve student health.

• Korean Journal of Food Science and Technology
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• v.33 no.4
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• pp.401-408
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• 2001

The ethanol extract and n-hexane fraction of Commiphora molmol Engl. showed minimum inhibitory concentration of 50 ppm and 25 ppm, respectively, on 5 strains of Listeria monocytogenes at $32^{\circ}C$. The purified substance, C3-3-2 fraction, was isolated by silica gel column and preparative thin layer chromatography from n-hexane fraction of Commiphora molmol Engl. The C3-3-2 fraction showed a strong bactericidal activity on 5 strains of L. monocytogenes at the concentration of 10 ppm in tryptic soy broth medium. At that concentration, the viable count was reduced $5{\sim}6$ log cycle from initial cell number. The n-hexane fraction of Commiphora molmol Engl. showed strong growth inhibition at the concentration of 25 ppm on Bacillus cereus and Staphylococcus aureus, at 50 ppm in broth on Salmonella enteritidis, and at 500 ppm on Vibrio parahaemolyticus. The purified antimicrobial substance, the C3-3-2 fraction, was identified as m-nonylphenol by on the basis of the $^1H-,\;^{13}C-NMR$ and EI/MS data. For the application test, the C3-3-2 fraction which was purely isolated from Commiphora molmol Engl. at 100 ppm were applied to minced Alaska pollack and ground beef at $32^{\circ}C$ and $5^{\circ}C$. The antimicrobial substances did not reduce L. monocytogenes ATCC 19113 at $32^{\circ}C$, while they reduced L. monocytogenes ATCC 19113 in viable number at $5^{\circ}C$. However, the antimicrobial effect of C3-3-2 fraction in food system was lower than that of broth condition.

• Applied Biological Chemistry
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• v.39 no.1
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• pp.54-59
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• 1996

Three strains of inhibitory lactic acid bacteria (No. 49, No. 61, No. 75) against Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, Escheirchia coli(ATCC33694) and Bacillus subtilis(ATCC6633) were isolated from kimchi, and then, identified to be Lactobacillus plantarum after examinations of their biological and physiological characteristics. To investigate a possible application of these three lactobacilli in milk fermentation industry, we made yogurts and then evaluated their ${\beta}-galactosidase$ activities at various; incubation time(from 24 hrs to 72 hrs). The result of experiment was that ${\beta}-galactosidase$ activities were reached maximum at 48 hrs and that reduced gradually with the lapse of time. And the ${\beta}-galactosidase$ activity of lactobacilli, and their viable cell counts at $37^{\circ}C$ for 2 hrs under various pH conditions were investigated. ${\beta}-galactosidase$ activities of 3 strains were reduced 50% at pH 3.5, but there were no remaining activities at pH 2.5, and pH 1.5, respectively. The frequency of the survival cell of lactobacilli in yogurt were $0.12{\sim}0.75%$ at pH 2.5, $$6.3{\times}10^{-5}{\sim}2.7{\times}10^{-3}% at pH 1.5, respectively, but there was no significant difference at pH 3.5. The values of original pH, titratable acidity as lactic acid, viscosity, and viable cells of yogurts were $4.08{\sim}4.30,\;1.05{\sim}1.25%,\;1,818{\sim}2,124\;cps\;and\;7.3{\times}10^8{\sim}3.0{\times}10^9\;cfu/m{\ell}$, respectively. To estimate buffer capacity of yogurt, the volume of 1.0 N HCl to 2 unit below original pH of yogurt($100\;m{\ell}$) was $11.98{\sim}13.02\;m{\ell}$ and the volume of 1.0N NaOH to 4 unit above original pH of yogurt($100\;m{\ell}$) was $10.82{\sim}12.86\;m{\ell}.

• Microbiology and Biotechnology Letters
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• v.45 no.3
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• pp.250-256
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• 2017

The prevalence of infections due to pathogenic bacteria such as Edwardsiella tarda, Streptococcus parauberis, and Photobacterium phosphoreum in fish farms in Jeju Island and their management by marine-derived biomaterials was studied. In this study, we isolated eight spices type of marine-derived biomaterials from four sea areas of Jeju Island. An antibiotic disc susceptibility test confirmed that the isolated marine-derived biomaterials showed weak resistance only to oxytetracycline and penicillin and sensitivity to the other antibiotics tested, and antimicrobial activity against fish pathogens with the inhibitory zone of 22 mm, 18 mm, and 19 mm for MD-02, MD-04, and MD-06 against E. tarda strains, respectively, and 19 mm, 22 mm, 30 mm, and 29 mm for MD-01, MD-02, MD-04, and MD-06 against S. parauberis strains, respectively, while all the marine-derived biomaterials showed antibacterial activity against P. phosphoreum. Among the eight biomaterials selected, Bacillus subtilis MD-02 displayed the greatest antibacterial activity against the three tested fish pathogens and also displayed susceptibility to antibiotics. The growth of Bacillus subtilis MD-02 was greatest with the carbon source, dextrine; nitrogen source, peptone; and mineral source, $MgSO_4{\cdot}7H_2O$. Hence, the present study confirmed that the isolate B. subtilis MD-02 from Jeju Island could be a potential antimicrobial agent against fish pathogens and a potential pharmacotherapeutic agent.