• Journal of Microbiology
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• v.39 no.4
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• pp.279-285
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• 2001

Vibrio parahaemolyticus is a halophilic bacterium associated with seafood gastroenteritis. An unusual strain of Kanagawa-positive urease producing Vibrio parahaemolyticus O1:K1 was isolated from the environment and identified . A polymerase chain reaction assay revealed that this strain harbored both the tdh and the genes. The urease from this strain was studied. Maximum urease production was induced in LB medium containing 0.2% urea, 0.5% glucose, 2% NaCl and pH 5.5 with 6h of culti-vation at 37$\^{C}$ under aeration. Purification of urease was achieved by the process of whole cell lysate, 65% ammonium sulphate precipitation, DEAE-cellulose ion exchange column chromatography, Sepharose CL-6B gel filtration and oxirane activated Sepharose 6B-urea affinity chromatography with 203 fold purification and 2.2% yield. Analysis of the purified enzyme by SDS-PAGE demonstrated the presence of the subunits with a molecular weight of 85kDa, 59kDa, 41kDa and the molecular weight for the native enzyme by nondenaturing PAGE and gel filtration chromatography was 255kDa. The purified urease was stable at pH 7.5 and the opeimal pH in HEPES buffer was 8.0 The enzyme was stable at 60$\^{C}$ for 2 h with a residual activity of 32% . The addition of 10$\mu$M if NiCl$_2$maintained stability for 30 min. The Km value of the purified enzyme was 35.6 mM in urea substrate. The TD$\_$50/(median toxic dose) of the purified urease was 2.5$\mu\textrm{g}$/ml on human leukemia cells.

• Mycobiology
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• v.30 no.1
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• pp.22-26
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• 2002

This study was carried out to investigate the possibility for seeds germination of Gastrodia elata using symbiotic fungi. Since seeds of G. elata are very small and lack an endosperm and other nutrients, their germination is difficult without requirement for external nutrients. Out of twenty six isolates collected from protocorms of G. elata and roots of native orchids inhabited in wild, two strains(H-2 and H-21) were observed to stimulate the seed germination of G. elata. The seed germination of G. elates was excellent on oak tree leaves medium. The optimal conditions for mycelial growth of symbiotic fungi were $25^{\circ}C$ and pH 6.0, respectively. The mycelial growth of H-2 strain was excellent on YMA medium, while H-21 was poor on PDA medium. In case of carbon sources, the mycelial growth of H-2 and H-21 was good on media supplemented with glucose and dextrin, respectively. Calcium nitrate was good for mycelial growth of H-2 strain as a nitrogen sources, whereas urea was effective to H-21 strain.

• Applied Biological Chemistry
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• v.43 no.3
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• pp.169-173
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• 2000

For the development of enzyme detergent capable of effectively washing at low temperature, a bacterium producing alkaline protease was isolated from soil samples, and properties of the enzyme were investigated. The selected strain was Gram negative, rod shape$(0.6{\sim}0.7{\times}1.3{\sim}2.6\;{\mu}m\;in\;size)$ and motile. It had the degradation activity of aesculin, gelatin and casein, and was catalase-positive. The cell wall components was meso-DAP, and G+C mole contents was 43.3%. From these results, the strain was identified as Acinetobacter sp. KN-27. The activity of alkaline protease by this strain peaked with 3,300 D.U/mL after 36 hours in the liquid culture at $40^{\circ}C$. The optimal pH and temperature of the enzyme were pH 9 and $60^{\circ}C$, respectively. Alkaline protease produced by Acinetobacter sp. KN-27 has shown two active bands on the electrophoresis of native gel.

• International Journal of Industrial Entomology and Biomaterials
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• v.35 no.2
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• pp.63-70
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• 2017

Native fluorescent pseudomonas bacteria were isolated from rhizosphere soil of mulberry and were evaluated against powdery mildew. In vitro conidial germination study showed significant (P<0.05) variation in conidial germination by bacterial strains Pf1 and Pf3. Mildew incidence was significantly varied due to treatment with various pseudomonas strains in vivo. Significantly (P<0.05) less mildew incidence was in plants treated with the bacterial strain Pf1 (9.11%) followed by Pf3 (13.48%) controlling 69.40% and 54.75% respectively compared with untreated control. Similarly, mildew severity was least (8.51%) in plants treated with strain Pf1 followed by Pf5 (9.23%) and Pf3 (9.72%) controlling the severity by 84.51%, 77.01% and 71.96% respectively compared with control. The bacterial strains significantly influenced biochemical constituents such as chlorophyll, protein and soluble sugar content of the mulberry leaf. Similarly, bacterial strains significantly increased the activity of the peroxidase (PO) and Polyphenol oxydase (PPO) activity from $7^{th}$ day up to the $28^{th}$ day after treatment. The strain Pf1, Pf3 and Pf5 exhibited a marked enhancement in the peroxidase at different periods of infection. Significant (P<0.01) negative correlation was found between powdery mildew severity with phenol content ($R^2=0.67$) as well as peroxidase ($R^2=0.92$) and polyphenol oxidase ($R^2=0.72$) activity thus confirms induction of systemic resistance in mulberry by pseudomonas bacteria. The study shows scope for exploration of rhizosphere fluorescent pseudomonas bacteria for induction of systemic resistance in mulberry to contain powdery mildew disease effectively.

• Journal of Applied Biological Chemistry
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• v.64 no.3
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• pp.225-236
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• 2021

The epipyrone (EPN) biosynthetic gene cluster of Epicoccum nigrum is composed of epnC, epnB, and epnA, which encode cytochrome P450 oxidase, glycosyltransferase, and highly reducing polyketide synthase, respectively. Gene inactivation mutants for epnA, epnB, and epnC were previously generated, and it was found that all of them were incapable of producing EPN and any of its related compounds. It was also reported that epnB inactivation abolished epnA transcription, generating ΔepnAB. This study shows that the introduction of native epnC readily restored EPN production in ΔepnC, suggesting that epnC is essential for polyketide release from EpnA and implies that EpnC works during the polyketide chain assembly of EpnA. Introduction of epnC promoter-epnA restored EPN production in ΔepnA. The ΔepnB genotype was prepared by introducing the epnA expression vector into ΔepnAB, and it was found that the resulting recombinant strain did not produce any EPN-related compounds. A canonical epnB inactivation strain was also generated by deleting its 5'-end. At the deletion point, an Aspergllus nidulans gpdA promoter was inserted to ensure the transcription of epnA, which is located downstream of epnB. Examination of the metabolite profile of the resulting ΔepnB mutant via LC-mass spectrometry verified that no EPN-related compound was produced in this strain. This substantiates that C-glycosylation by EpnB is a prerequisite for the release of EpnA-tethered product. In conclusion, it is proposed that cytochrome P450 oxidase and glycosyltransferase work in concert with polyketide synthase to generate EPN without the occurrence of any free intermediates.

• Korean Journal of Poultry Science
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• v.48 no.4
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• pp.217-225
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• 2021

This study compares the physicochemical characteristics and fatty acid composition of three Korean native chickens and broilers. Ten whole raw broiler chickens and ten each from the three Korean native chickens (KNCs), Hanhyup 3 (HH3), Woormatdak 1 (WRMD1), and Woormatdak 2 (WRMD2), were purchased from the meat market. Their breast and thigh meat were used as samples. The proximate composition, pH, color, water-holding capacity (WHC), shear force, collagen content, and fatty acid composition were determined. In breast meat, the moisture content of HH3 (74.94%) and WRMD1 (74.74%) was lower than that of the broilers (77.1%, P<0.05). No significant difference was found in crude protein, lipids, and ash contents. The crude fat from thigh meat from HH3 and WRMD2 was lower than that of broilers (P<0.05). The redness of WRMD1 was the highest in both breast and thigh meat (P<0.05). The WHC of the breast meat of WRMD1 was lower than that of HH3 and WRMD2. In thigh meat, the WHC of the broilers was significantly higher than that of the KNCs. In breast meat, the shear force of WRMD2 was significantly lower than that of the broilers, HH3, and WRMD1, while no significant difference was found in thigh meat. The collagen content and arachidonic acid levels of the KNCs were significantly higher than those of the broilers for breast and thigh meats. No significant differences were observed among the KNCs. This result can be used to improve the quality of KNC but further studies on the bioactive compounds, taste, and volatile compounds of KNCs are required.

• Journal of Microbiology
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• v.45 no.2
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• pp.179-184
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• 2007

Pasteurella multocida, a Gram-negative facultative anaerobic bacterium, is a causative animal pathogen in porcine atrophic rhinitis and avian fowl cholera. For the development of recombinant subunit vaccine against P. multocida, we cloned and analyzed the gene for outer membrane protein H (ompH) from a native strain of Pasteurella multocida in Korea. The OmpH had significant similarity in both primary and secondary structure with those of other serotypes. The full-length, and three short fragments of ompH were expressed in E. coli and the recombinant OmpH proteins were purified, respectively. The recombinant OmpH proteins were antigenic and detectable with antisera produced by either immunization of commercial vaccine for respiratory disease or formalin-killed cell. Antibodies raised against the full-length OmpH provided strong protection against P. multocida, however, three short fragments of recombinant OmpHs, respectively, showed slightly lower protection in mice challenge. The recombinant OmpH might be a useful vaccine candidate antigen for P. multocida.

• Korean Journal of Veterinary Research
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• v.37 no.4
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• pp.875-882
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• 1997

To produce the recombinant fibronectin-binding protein(FnBP) for development of subunit vaccine against Staphylococcus aureus. The fnbp gene was amplified from the chromosomal DNA of S aureus KNU 196 strain using the polymerase chain reaction, and cloned into pGEX-4T-2. Then, the recombinant FnBP fused with glutathione-S-transferase was produced in E coli, purified by affinity chromatography, and identified its antigenicity and immunogenicity by Western blot. The recombinant FnBP produced in this study is considered to have the same property of native FnBP purified from S aureus, and is expected to be useful as a candidate for S aureus subunit vaccine.

• Proceedings of the KSAR Conference
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• 2002.06a
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• pp.16-16
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• 2002

The egg productivity of the chick is represented by the number of egg produced, egg weight, and sexual maturity, which are regulated by various endocrine factors. Although there were some reports that insulin-like growth factor-I (IGF-Ⅰ) affected egg production, studies on any correlation between IGFs and egg productivity were not reported in poultry. The objectives of the present study were to examine the IGFs profile and egg productivity in both KNOC and laying hen (Saver) and to investigate the relationship of IGFs with egg productivity. Whole blood was collected every 10 wk until 60 wks. (omitted)

• Microbiology and Biotechnology Letters
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• v.17 no.6
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• pp.587-592
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• 1989

A strain of alkalophilic Bacillus sp. YS-309 capable of producing large amount of $\beta$-galactosidase has been isolated from soil sample. Intracellular $\beta$-galactosidase was purified 6.9 folds by procedures including ammonium sulfate precipitation, DEAE-cellulose chromatography, gel-filtration, DEAE-Sephadex A-50 chromatography with over-all yield of 17.8%. The molecular weight of native enzyme was 205, 000 by HPLC, and SDS-polyacrylamide gel electrophoresis showed that the enzyme consisted of 4 identical subunits with a molecular weight of 56, 000.