• Proceedings of the Botanical Society of Korea Conference
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• 2002.04a
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• pp.62-72
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• 2002

This study has investigated the biosynthesis and function of the heavy metal binding peptides, the phytochelatins, in plants. PCs are synthesised enzymatically from glutathione by the enzyme PC synthase in the presence of heavy metal ions. Using Arabidopsis thaliana as a model organism cadmium-sensitive, phytochelatin-deficient mutants have been isolated and characterised in previous studies. The cadl mutants have wildtype levels of glutathione, are PC deficient and lack PC synthase activity. Thus, the CADl gene has been proposed to encode PC synthase. The CADl gene was isolated by a positional cloning strategy The gene was mapped and a candidate identified. Each of four cadl mutants had a single base pair change in the candidate gene and the cadmium-sensitive, cadl phenotype was complemented by the candidate gene. This demonstrated the CADl gene had been cloned. A homologous gene in the fission yeast, Schizosaccharomyces pombe was identified through database searches. A targeted-deletion mutation of this gene was constructed and the mutant, like cadl mutants of Arabidopsis, was cadmium-sensitive and PC-deficient. A comparison of the redicted amino acid sequences reveals a highly conserved N-terminal region Presumed to be the catalytic domain and a variable C-terminal region containing multiple Cys residues proposed to be involved in activation of the enzyme by metal ions. Similar genes were also identified in animal species. The Arabidopsis CADl/AtPCSl and S. pombe SpbPCS genes were expressed in E. coli and were shown to be sufficient for glutathione-dependent, heavy metal activate PC synthesis in vitro, thus demonstrating these genes encode PC synthase enzymes. Using RT-PCR, AtPCSl expression appeared to be independent of Cd exposure. However, at higher levels of Cd exposure a AtPCSl-CUS reporter gene construct appeared to be more highly expressed. Using the reporter gene construct, AtPCSl was expressed most tissues. Expression appeared to be greater in younger tissues and same higher levels of expression was observed in some regions, including carpels and the base of siliques. AtPCS2 was a functional gene encoding an active PC synthase. However, its Pattern of expression and the phenotype of a mutant (or antisense line) have not been determined. Assuming the gene is functional then it has clearly been maintained through evolution and must provide some selective advantage. This implies that, at least in some cells or tissue, it is likely to be the dominant PC synthase expressed. This remains to be determined

• Korean Journal of Veterinary Research
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• v.39 no.3
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• pp.523-530
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• 1999

Sal enteritidis thin fimbriae, SEF14, were found to be restricted to the predominantly poultry-associated members of the Salmonella serogroup D1 that are considered as the important pathogens in poultry industry. SefA together with sefB and sefC encode the proteins involved in SEF14 biosynthesis. In order to develop the rapid and specific detection methods for Salmonella serogroup D1, a PCR technique for the amplification of sefA gene was established, and its specificity and sensitivity were investigated with various microorganisms. The bacterial genomic DNA was extracted by colony-picking and rapid boiled-lysate technique. In comparison of Sef I and Sef II primers used in the PCR, Sef I primer for sefA gene of 513bp showed higher specificity than that of Sef II. The established PCR was as sensitive as to detect 1pg of Sal enteritidis DNA. When 73 strains in 28 genera including the reference strains and the field isolates of various Salmonella serotypes, Bacillus subtilis, Bordetella bronchisepdca, E coli, Listeria spp., Micrococcus luteus, Rhodococcus equi, Staphylococcus spp., Streptococcus spp., Vibrio parahemolyticus, Yersinia spp. were studied, the established PCR yielded specifically positive results with only Salmonella serogroup D1. The results suggested that the PCR for sefA gene could be a potential candidate among the specific detection methods for Salmonella serogroup D1.

• Korean Journal of Veterinary Research
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• v.40 no.2
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• pp.345-360
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• 2000

The effects on CTC (T1) and CTC, Sulfathiazole and Penicillin Combination (T2) medication in feed through one life cycle of pigs, namely, weaning, mating, farrowing, lactation, growing, finishing and slaughter, were tested under local condition. In sow phase, productivity and the number of microflora in urine before and after medication of CTC were studied and average daily gain and feed conversion rate were checked during growing and finishing period. All pigs reached at 155 days old were slaughtered for pathological examination. 1. Litter size, farrowing rate and survival rate at birth were improved by CTC medication from weaning to 21th day after mating and mortality of piglet at weaning, 25 days after farrowing, was reduced in the CTC medication group, but no siginificant. 2. The number of microflora in the sow urine was changed with the medication at 200ppm of CTC in feed. In particular, the number of E coli, Samonella and Staphylococci were reduced by CTC medication. 3. The average daily gain and feed conversion rate of grower and finisher pigs was improved significantly in both treated groups, most in the high level CTC (T1) medicated group and was lowest in the control group. 4. The number of infected lungs was reduced not significant by both treatments (as % pneumonic lesions Co 66.7%, T1 47.1%, T2 31.4%) and the severity of lung lesions was significantly reduced by both high level of CTC and CTC combination medication in feed. 5. Although there were no statistical differences in atrophic rhinitis based on turbinate scores among the 3 groups, the number of mild and moderate (Grade 2 and 3) infections was higher in the control group (9/36) than in the treated groups (T1 2/34 & T2 4/35).

• Molecules and Cells
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• v.24 no.2
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• pp.276-282
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• 2007

Protein phosphorylation is one of the major mechanisms by which eukaryotic cells transduce extracellular signals into intracellular responses. Calcium/calmodulin ($Ca^{2+}/CaM$)-dependent protein phosphorylation has been implicated in various cellular processes, yet little is known about $Ca^{2+}/CaM$-dependent protein kinases (CaMKs) in plants. From an Arabidopsis expression library screen using a horseradish peroxidase-conjugated soybean calmodulin isoform (SCaM-1) as a probe, we isolated a full-length cDNA clone that encodes AtCK (Arabidopsis thaliana calcium/calmodulin-dependent protein kinase). The predicted structure of AtCK contains a serine/threonine protein kinase catalytic domain followed by a putative calmodulin-binding domain and a putative $Ca^{2+}$-binding domain. Recombinant AtCK was expressed in E. coli and bound to calmodulin in a $Ca^{2+}$-dependent manner. The ability of CaM to bind to AtCK was confirmed by gel mobility shift and competition assays. AtCK exhibited its highest levels of autophosphorylation in the presence of 3 mM $Mn^{2+}$. The phosphorylation of myelin basic protein (MBP) by AtCK was enhanced when AtCK was under the control of calcium-bound CaM, as previously observed for other $Ca^{2+}/CaM$-dependent protein kinases. In contrast to maize and tobacco CCaMKs (calcium and $Ca^{2+}/CaM$-dependent protein kinase), increasing the concentration of calmodulin to more than $3{\mu}M$ suppressed the phosphorylation activity of AtCK. Taken together our results indicate that AtCK is a novel Arabidopsis $Ca^{2+}/CaM$-dependent protein kinase which is presumably involved in CaM-mediated signaling.

• Korean Journal of Microbiology
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• v.46 no.3
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• pp.270-277
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• 2010

In order to manufacture Bacillus cereus-free fermented soybean products, an antimicrobial agentproducing isolate against B. cereus was obtained from 150 traditionally fermented soybean products. The morphological and biochemical tests and the phylogenetic relationship among 16S rRNA gene sequences indicated that the isolate named as the strain SCK 121057 was most closely related to Bacillus licheniformis. The B. licheniformis isolate began to produce the antimicrobial agent after 48 h of incubation. The agent was nonproteinaceous and insensitive to heat, long term storage and protease K. Electron microscopic observation indicated that the agent attacked the membrane of B. cereus, leaving the ghost cell. The isolate inhibited growth of B. subtilis, Lactobacillus brevis and various types of pathogenic strains including Escherichia coli, E. faecalis, Micrococcus luteus, Staphylococcus aureus, Aspergillus flavus, A. ochraceus, and A. parasiticus as well as B. cereus. After coinoculation of B. licheniformis SCK 121057 and B. cereus in the ratio (as the basis of CFU/g sample) of 10 to 1 on the surface of cooked soybeans, cell numbers of B. cereus had been dramatically reduced after 31 days of incubation compared to those of single inoculation of B. cereus.

• Journal of Microbiology and Biotechnology
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• v.30 no.11
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• pp.1720-1728
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• 2020

We have previously characterized AprESJ4, the major fibrinolytic enzyme from Bacillus subtilis SJ4 (Yao et al., 2019). During that study, we observed a 68 kDa protein with fibrinolytic activity. In this study, we cloned the gene (vprSJ4) encoding the 68 kDa protein, a mature Vpr and minor protease secreted by Bacillus species. vprSJ4 encodes a preproenzyme consisting of 810 amino acids (aa) including signal sequence (28 aa) and prosequence (132 aa). The mature enzyme (650 aa) has a predicted molecular weight of 68,467.35. Unlike Vprs from other B. subtilis strains, VprSJ4 has 4 additional amino acids (DEFA) at the C-terminus. vprSJ4 was overexpressed in Escherichia coli. PreproVprSJ4 was localized in inclusion bodies, and subjected to in vitro renaturation and purification by an affinity column. SDS-PAGE and western blot showed that autoprocessing of preproVprSJ4 occurred and 68 kDa and smaller proteins were produced. The optimum pH and temperature of the recombinant VprSJ4 were pH 7.0 and 40℃, respectively. Kinetic parameters of recombinant VprSJ4 were measured by using an artificial substrate, N-succinyl-ala-ala-pro-phe-p-nitroanilide. Coexpression of vprSJ4 and aprESJ4 using pHY300PLK increased the fibrinolytic activity a further 117% when compared with aprESJ4 single expression using the same vector in B. subtilis WB600.

• Korean Journal of Food Science and Technology
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• v.37 no.1
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• pp.11-14
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• 2005

White radish was minimally processed, packed with air, $CO_{2}$ (100%), and $CO_{2}/N_{2}$ (25/75%), and irradiated at 0, 1, and 2 kGy, and its microbiological quality and pH were investigated during storage for 2 weeks at $4^{\circ}C$. Irradiation significantly reduced total aerobic, coli-form, and lactic acid bacteria counts. Modified atmosphere packaging (MAP) enhanced microorganism control during storage. Acidity decreased by MAP but was restored during storage, Irradiation did not affect sample pH. Results show irradiation at 2 kGy combined with MAP can enhance microbiological safety and quality of minimally processed radish.

• Journal of the East Asian Society of Dietary Life
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• v.27 no.5
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• pp.576-582
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• 2017

This study evaluated the microbial quality of dried and powdered foods during storage with increased humidity because of climate change. Five types of dried and powdered foods (dried shredded squid, wheat flour, Sunsik, red pepper powder, and roasted sesame seed) were stored at different relative humidities (RH 23%, 43%, 68%, 85%, and 100%) and changes in water activity and microbial populations were measured during storage at $35^{\circ}C$ for 15 days. The results revealed that water activity values of dried and powdered foods were significantly increased during storage when samples were stored at RH 85 and 100%. In addition, levels of total mesophilic bacteria, yeast, and mold were significantly increased after storage for 6 days or 9 days at RH 85% and 100%. However, levels of Escherichia coli and coliform did not increase significantly during storage. Based on these findings, dried and powdered foods should not be stored at high RH because the increased water activity enables microbial growth.

• Journal of the Korean Society of Food Science and Nutrition
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• v.25 no.5
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• pp.761-767
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• 1996

To lessen the initial level of microorganisms, electrolyzed acid-water was used as washing and brine water in the manufacturing process. On the washing and salting processes, application of electrolyzed acid-water showed a possibility to lessen the microorganism level of Chinese cabbage effectively. Microbial level of Chinese cabbage was reduced to about 1/4 level by salting and washing process with electrolyzed acid-water while Chinese cabbage salted with tap water increased to about 1.7 times. And no coliform and E. coli were detected. However significant differences between seasoning mixtures prepared with electrolyzed acid-water and with tap water were not observed in microbial levels. Relatively low level of total count in kimchi prepared with electroyzed acid-water was kept until 15 days of fermentation at $10^{\circ}C.$ Any significant difference between them was not observed after 20 days of fermentation. pH and acidity were showed the same tendencies as microbial count.

• Food Science and Preservation
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• v.13 no.5
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• pp.629-633
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• 2006

Pinus rigida Miller leaf extract (PRLE) showed antimicrobial activity remarkably against food pathogenic and spoilage bacteria at concentrations of $100{\sim}250{\mu}g/mL$. Alcohol-soluble PRLE had higher antimicrobial activity against Staphylococcus aureus and E-coli than any other-soluble PRLE such as butanol, ethyl acetate, ether and water. As PRLE concentration increased alcohol-soluble PRLE increased the remarkable inhibitory zone of microbial growth on the microbial media. PRLE showed good stability against temperature and pH in the range of $40{\sim}150^{\circ}C$ and $4{\sim}11$, respectively. This may indicate that PRLE can be a potential anti-microbial agent for industrial application. In addition, SEM of Listeria monocytogenes suggested that it antimicrobial component would perturb the functions of microbial cell membranes synergistically. In the feeding experiment the formaldehyde content in the serum of formalin-fed and PRLE-treated me decreased remarkably due to the lysis of formaldehyde and the rate of hemoglobin biosynthesis was recovered to the orignal state within a short breeding time.