• KSBB Journal
• /
• v.16 no.6
• /
• pp.562-567
• /
• 2001

Aspergillus niger LK harboring the enantioselective epoxide hydrolase (EHase) activity was isolated, and enantioselectivity of EHase was tested for various racemic aromatic epoxides. The gene encoding epoxide hydrolase was cloned from cDNA library generated by reverse transcriptase-polymerase chain reaction of the isolated total mRNA. Sequence analysis showed that the cloned gene encodes 398 amino acids with a deduced molecular mass of 44.5 kDa. Database comparison of the amino acid sequence reveals that it is similar to fungal EHase, whereas the sequence identity with bacterial EHase is very low. Recombinant expression of the cloned EHase in Escherichia coli BL21 yielded an active EHases, which can offer a potential biocatalyst for the production of chiral epoxides.

• The Journal of the Korean Society for Microbiology
• /
• v.35 no.1
• /
• pp.69-76
• /
• 2000

Members of the genus Acinetobacter are recognized as newer pathogens of the nosocomial infection with an increasing frequency in recent years. Strains that belonged to A. calcoaceticus A. baumannii complex (genomic species 1, 2, 3, and 13TU) were major groups associated with nosocomial infection. Phenotypic identification was unreliable and laborious method to classify Acinetobacter strains into 19 genomic species. Rapid and reliable identification of clinical isolates is essential to diagnosis and epidemiology of Acinetobacter. We investigated the suitability of amplified ribosomal DNA restriction analysis (ARDRA) to identify genomic species of 131 Acinetobacter isolates. The 16S rRNA genes (ribosomal DNA) were enzymatically amplified and the amplified PCR products were restricted independently with the enzymes, AluI, CfoI, and MboI. Genomic species of Acinetobacter was classified by the combinations of restriction patterns. The analysis was showed that restriction profiles were characteristic for each genomic species. One hundred fourteen isolates were identified as A. baumannii, twelve were identified as genomic species 13TU, and one was identified as genomic species 3. Four isolates were found to be unknown organisms. All of the isolates which were identified to A. baumannii by phenotypic tests were completely discriminated into A. baumannii and genomic species 13TU by ARDRA. This study demonstrates that ARDRA is a rapid and simple techniques for the identification of Acinetobacter species according to the genomic species.

• Korean journal of applied entomology
• /
• v.51 no.4
• /
• pp.377-382
• /
• 2012

This study investigated the occurrence of sweet potato whitefly, Bemisia tabaci affecting cucumber, eggplant and red pepper, as well as sweet potato species, and its response to insecticides in Gyeonggi province from 2010 to 2011. Sweet potato whitefly is widespread throughout the southern part of Gyeonggi province. Most regional populations of B. tabaci belong to biotype Q having been reported in the south Korea since 2005, but in Goyang mixed populations of two biotypes (B and Q) were found. Survey results of Tomato Yellow Leaf Curl Virus (TYLCV) disease that was vectored by B. tabaci indicated that this virus disease was not spread throughout the Gyeonggi province. Biotype Q of B. tabaci was found to be resistant to neonicotinoid insecticides, whereas biotype B was highly susceptible to them.

• Journal of Life Science
• /
• v.24 no.6
• /
• pp.694-699
• /
• 2014

Indigenous microorganisms play decisive roles in biodegradation. In this study, eighty strains of hydrocarbon-degrading microbes were isolated from Incheon Bay. Among them, 12 strains were selected by an oil film collapsing method. The bacterial strain 'Incheon9' was eventually selected based on its relatively higher lipase and emulsification activities, and was identified as Acinetobacter sp. (NCBI accession code: KF54854). The optimum condition for the growth and emulsification activity of Acinetobacter sp. Incheon9 was $20^{\circ}C$, pH 7, and 1% NaCl. The optimum time for the best production of biosurfactant was 72 hrs. The oil degradation ability of Acinetobacter sp. Incheon9 was investigated by measuring the residual oils in the culture medium by gas chromatography (FID). This research provides foundational data for eco-friendly environmental remediation by microorganisms.

• Korean Journal of Veterinary Research
• /
• v.51 no.2
• /
• pp.107-115
• /
• 2011

Theileria (T.) buffeli (formerly T. sergenti/T. orientalis) is the major hemo-protozoan distributed in the Far East Asian countries such as Korea, China and Japan. It is responsible for the clinical symptoms of anorexia, ateliosis, anemia, fever and icterus. It also causes abortion and sudden death under severe cases, resulting in economic losses for many livestock farms. The objective of this study was to analyze the genetic diversity of the major surface protein (Msp) gene in T. buffeli in Holstein in Korea, and we characterized the association of the diversification of the Msp gene and its relationship with the pathogenicity of Theileria. For this, complete blood counts and Theileria PCR sequence analysis were performed from 57 Holstein in Jeju Island. A total of 26 PCR positive Holstein (16 anemic and 10 non-anemic) were then randomly selected based on 18s rRNA sequence typing of the Theileria Msp gene. The DNA sequence of the T. buffeli Msp gene in Holstein showed 99.0%, 99.2%, 99.9%, 99.5%, 98.7%, 98.4% and 98.4% homology with T. sergenti, Theileria spp., T. sergenti, Theileria spp., Theileria spp., Theileria spp. and Theileria spp., respectively. The result showed a genetic variation of 57.7% (type I), 3.8% (type II), 15.4% (type III), 7.7% (type IV), 13.5% (type V) and 1.9% (type VI). Type I is the most frequent type in both anemic and non-anemic Holstein while type II was found in only non-anemic Holstein. This results of our study help confirm the diversity of Msp gene types and demonstrate that the gene type distribution of Msp genes varies among Theileria-infected Holstein in Jeju Island.

• Journal of Life Science
• /
• v.24 no.10
• /
• pp.1110-1117
• /
• 2014

Tidal flats are continuously contaminated by human activities. This study assessed the bioremediation efficiency of tidal flat soil using microcosm reactors and microorganisms originating from the tidal area. We screened 135 bacterial strains that produce extracellular enzymes from the tidal area located in the North port of Incheon bay. Two bacterial strains (Pseudoalteromonas sp. and IC35 Halothiobacillus neapolitanus IC_S22) were selected and used in the microcosm reactors, which were specially designed to functionally mimic the ecological conditions of the tidal flats. Pseudoalteromonas sp. IC35 was selected based on its relatively high activity of the enzymes amylase, cellulose, lipase, and protease. Halothiobacillus neapolitanus IC_S22 was selected for oxidation of sulfur. The M1 and M2 microcosm reactors were operated by continuous feeding of seawater under the same conditions, but M2 was first inoculated with Pseudoalteromonas sp. IC35 before the seawater feeding. The initial COD in both the M1 and M2 microcosm reactors was 320 mg/l. The final COD was 21 mg/l (M1) and 7 mg/l (M2). The M3 and M4 microcosm reactors were operated by continuous feeding of seawater under the same conditions, but M4 was first inoculated with H. neapolitanus IC_S22. The initial sulfate concentration in both the M3 and M4 microcosm reactors was 660 mg/l, and the maximum sulfate concentration was 1,360 mg/l (M3) and 1,600 mg/l (M4).

• Korean Journal of Microbiology
• /
• v.48 no.2
• /
• pp.134-140
• /
• 2012

Asian dust storms originating in the arid desert of China and Mongolia usually occur from late winter through spring, and more than one million tons of dust per year is transported to the Korean Peninsula by the prevalent westerly winds. We supposed that these dust particles could include bioaerosols and act as carriers of microorganisms. In order to clarify the dynamics of microorganisms moving with these particles, the concentration and composition of microorganisms associated with settled particles were compared between samples collected during Asian dust events and those under non-dust periods. From February to April 2008, settled dust particles were collected at one location in Ulsan using rainfall meter of 200 mm diameter. During this period, there was one Asian dust event in Ulsan. The bacterial concentrations were higher in samples collected during Asian dust event than those under non-dust period, whereas fungal concentrations were rather similar regardless of the Asian dust event. We analyzed 16S rRNA gene sequences of 45 bacterial isolates obtained from the settled particle samples. These isolates belonged to either genus Bacillus or genus Streptococcus and were tentatively identified as B. amyloliquefaciens, B. aryabhattai, B. atrophaeus, B. licheniformis, B. megaterium, B. methylotrophicus, B. pumilus, B. sonorensis, B. subtlis, B. vallismortis, S. epidermidis, and S. succinus. In cases of fungal isolates, genera such as Mucor, Alternaria, Cladosporium, and Aspergillus were tentatively identified from samples collected at both Asian dust and non-Asian dust periods. It appears that endospore-forming bacteria such as Bacillus sp. rather than fungal spores are more likely to be associated with Asian dust particles.

• Research in Plant Disease
• /
• v.22 no.4
• /
• pp.227-235
• /
• 2016

Plant disease caused by root-knot nematode is a major problem in crop production. Using of chemical pesticides, one of the most efficient methods to control nematodes, have raised issues in toxicity to humans and animals and environmental pollution. In this study, to select actinomycete strains that have potential to serve as a microbial agent for control of nematodes, we investigated nematicidal activity of culture broth from 670 Streptomyces isolates. A culture filtrate of KRA15-528 isolate that was identified as S. flavogriseus on the basis of 16S rRNA sequence analysis, showed strong nematicidal activity against second stage of juveniles of Meloidogyne incognita and inhibited egg hatching; exposure to 10% of culture filtrate resulted in 71% juvenile mortality at 48 hours afters treatment and suppressed egg hatching by 54% at 9 days after treatment. When the KRA15-528 culture filtrate was partitioned with ethyl acetate and butanol, ethyl acetate layer exclusively showed strong activity; 91%, 53%, 30% of mortality at 1,000, 500, $250{\mu}g/ml$, respectively. Additionally, the culture filtrate suppressed gall formation on cucumber plant by M. incognita with no phytotoxicity. These results suggest that S. flavogriseus KRA15-528 has potential to serve as a microbial nematicide for the control of root-knot nematode disease.

• Journal of Applied Biological Chemistry
• /
• v.62 no.3
• /
• pp.287-292
• /
• 2019

Pseudomonas tolaasii causes brown blotch disease on the oyster mushroom (Pleurotus ostreatus). Various pathogenic strains of P. tolaasii were isolated and divided into three subtypes, $P1{\alpha}$, $P1{\beta}$, and $P1{\gamma}$. For phage therapy, bacteriophages against to these subtype strains were applied to mushroom cultivation and very successful to prevent from the disease. In this study, bacteriophages were isolated against the representative strains of subtype pathogens and their polyclonal antibodies were synthesized to investigate structural relationship among capsid proteins of phages. Phage preparations over $10^{10}pfu/mL$ were injected to rabbit thigh muscle and polyclonal antibodies were obtained after three times of boost injection. Titers of the antibodies obtained were over $2{\times}10^7Ab/mL$ for the phage ${\phi}6264$, $1{\times}10^6Ab/mL$ for the phage ${\phi}HK2$, and $1{\times}10^7Ab/mL$ for the phage ${\phi}HK19$ and phage ${\phi}HK23$. High specific activities were observed between antibodies and the corresponding bacteriophages. Some cross-reactivities between the antibodies and non-corresponding bacteriophages were also measured. Antibody $Ab{\phi}6264$ inactivated all phages of $P1{\alpha}$ subtype and only phage ${\phi}HK16$ among $P1{\beta}$ subtype phages. Antibody $Ab{\phi}HK23$ of $P1{\gamma}$ subtype neutralized all phages of $P1{\beta}$ subtype as well as the phage ${\phi}HK23$, showing the widest phage-inactivation range. When the structural-similarity studies of phages were investigated by using phage antibodies, closeness obtained by phylogenetic analysis of 16S rRNA genes of pathogenic strains were quite different from that of polyclonal antibody-specific structural similarity of phage capsid proteins. In conclusion, there is weak correlation between the host strain specificity of bacteriophage and its capsid structural similarity measured by phage antibodies.

• Journal of Life Science
• /
• v.29 no.8
• /
• pp.861-870
• /
• 2019

This study was undertaken to establish optimum medium compositions for cost-effective mannitol production by Leuconostoc mesenteroides SRCM201425 isolated from kimchi. L. mesenteroides SRCM21425 from kimchi was selected for efficient mannitol production based on fructose analysis and identified by its 16S rRNA gene sequence, as well as by carbohydrate fermentation pattern analysis. To enhance mannitol production by L. mesenteroides SRCM201425, the effects of carbon, nitrogen, and mineral sources on mannitol production were first determined using Plackett-Burman design (PBD). The effects of 11 variables on mannitol production were investigated of which three variables, fructose, sucrose, and peptone, were selected. In the second step, each concentration of fructose, sucrose, and peptone was optimized using a central composite design (CCD) and response surface analysis. The predicted concentrations of fructose, sucrose, and peptone were 38.68 g/l, 30 g/l, and 39.67 g/l, respectively. The mathematical response model was reliable, with a coefficient of determination of $R^2=0.9185$. Mannitol production increased 20-fold as compared with the MRS medium, corresponding to a mannitol yield 97.46% when compared to MRS supplemented with 100 g/l of fructose in flask system. Furthermore, the production in the optimized medium was cost-effective. The findings of this study can be expected to be useful in biological production for catalytic hydrogenation causing byproduct and additional production costs.