• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.445-448
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• 2002

The Pseudomonas fluorescens KCTC 1767, a selected and identified as potential candidate for stereo-specific resolution of rac-ketoprofen ethyl ester, was systematically investigated in order to induce the high level expression and detailed characterization of the expressing enzyme esterase. We cloned the esterase gene from chromosomal DNA of Pseudomonas fluorescens KCTC 1767 by PCR with two synthetic primers that desinged for simple purification. The recombinant esterase from Pseudomonas fluorescens KCTC 1767 exibited a high conversion rate and enantioselectivity to the (S)-ketoprofen ethyl ester as expected. The enzyme was easily purified to homogeniety by using a metal chelating affinity chromatography as a protein with poly histidine taq, and thus obtained 0.6 mg of protein from a 100 mL culture broth in a single step. The purified enzyme was steadily stable at the pH range from 7.0 to 10. The activity was also retained to be about 70% after the preincubation at $40^{\circ}C$ but over $50^{\circ}C$ lost the activity completely. The molecular mass of the esterase was estimated to be about 43 kDa on SDS-PAGE, and an identical result was also shown in gel filteration chromatography. The specific activity was calculated 27 mM/mg-protein/min by using the rac-ketoprofen ethly ester as a substrate.

• Journal of Microbiology and Biotechnology
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• v.1 no.2
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• pp.102-110
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• 1991

Alkalophilic Bacillus sp. YJ-451, which was isolated from soil at several area in Korea, produced a novel type of bacteriolytic enzyme (cell wall peptidoglycan hydrolase) extracellulary. The cell wall hydrolytic activity was identified as a clear zone on sodium dodecyl sulfate polyacrylamide gel electrophoresis containing 0.2% (w/v) cell wall of Bacillus sp. as substrate. This enzyme was successively purified 66 fold with 3.2% yield in culture broth by ammonium sulfate precipitation, CM-cellulose column chromatography, and gel filtration, followed by hydroxylapatite column chromatography. The molecular weight of the purified enzyme was estimated to be 27,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis and gel filtration column chromatography. The optimum pH and temperature for the activity of the enzyme were pH 10.0 and $50^{\circ}C$, respectively. The enzyme was stable between pH 5.0 and 10.0 and up to $40^{\circ}C$. Among the microorganisms used in this experiment the enzyme was active against most of gram negative strains and the genus Bacillus such as B. megaterium, B. licheniformis, B. circulans, B. pumilus, B. macerans, B. polymyxa. The release of dinitrophenylglutamic acid but not reducing group from cell wall peptidoglycan digested by the enzyme suggested that the enzyme is a kind of peptidase which hydrolyzes the peptide bond at the amino group of D-glutamic acid in the peptidoglycan.

• Microbiology and Biotechnology Letters
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• v.32 no.4
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• pp.366-370
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• 2004

Four chitinase producing bacteria, Arthrobacter nicotinae CH4, Arthrobacter nicotinae CHI3, Arthrobacter sp. CH5 and Micrococcus sp. CH3, were isolated from small crabs and shrimps. We investigated the optimum medium condition for the production of enzyme and high cell mass. The preferable medium composition was as follows: colchitin 0.1 %(w/v), glycerol 0.25%(w/v) and yeast extract 0.05%(w/v) in minimal midium ($K_{2}HPO_{4}$ 0.7 g/l, $KH_{2}PO_{4}$ 0.3 g/l, $MgSO_{4}{\cdot}5H_{2}O$ 0.5 g/l, $FeSO_{4}}{\cdot}7H_{2}O$ 0.01 g/l, $ZnSO_{4}$ 0.001 g/l, $MnCI_2$ 0.001 g/l, pH 7.0). This cell culture medium could be used directly as sample for measuring chitinase activity. Because it hardly conreducing sugar such as glucose (blank value=0), the detected reducing sugar can be considered as a chitinase reaction product. The results can be used for easy preparation method for determination of enzyme activity and analysis of enzyme-substrate reaction in step of screening of chitinase producing bacteria.

• Microbiology and Biotechnology Letters
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• v.22 no.5
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• pp.447-452
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• 1994

The strain SL-387 which produces new inhibitors of aminopeptidase M, MR-387A and B, was isolated from a soil sample. The strain has branched substrate mycelia, from which aerial hyphae develop in the form of open spirals. Spore surface is smooth. Melanoid and soluble pigme- nts were observed. The isolate contains LL-diaminopimelic acid in its cell wall hydrolysate, and has no pectinolytic activity. The strain SL-387 is closely related to Streptomyces griseoruber and S. naganishii, but is different from these strains in some cultural and physiological characteristics. This strain was, therefore, designated as Streptomyces sp. SL-387. The effects of several carbon and nitrogen sources on the production of the inhibitor were examined. Among them, glucose, galactose, mannose, and xylose were effective as a carbon source and soybean meal, soytone, fish meal, and gluten meal were effective as a nitrogen source. The maximum peak of the inhibitor production in jar fermentor was obtained on the fifth day of culture.

• International Journal of Industrial Entomology and Biomaterials
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• v.13 no.2
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• pp.57-61
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• 2006

In this study, morphology of perithecia, asci, ascospores, etc. of C. sphecocephala were examined for its telemorphic characteristics. Its colony grew up to 32 mm in diameter on potato dextrose agar (PDA) for 30 days under the condition of $24{\pm}1^{\circ}C$. PDBLA and PDBAA media were selected as optimal media for C. sphecocephala, on which the growth was 1.5 times as fast as on PDA medium. Moreover, PDBLA medium induced successfully the synnemata of anamorphic state. C. sphecocephala was able to be proliferated in vitro on both larva and adult of honeybee drone as its substrate. After inoculated onto the drone larva, it produced mycelium at $24{\pm}1^{\circ}C$, with the maximum yield up to $67{\pm}3mg$ on the $50^{th}$ day.

• Korean Journal of Microbiology
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• v.21 no.3
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• pp.143-148
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• 1983

The occurrence of PZ-peptidase in Streptococcus sanguis and other oral bacteria was investigated utilizing washed whole cells as the enzyme source and PZ-pentapeptide as its substrate. Under the culture conditions employed in the present study. Streptococcus sanguis strains, fresh isolates as well as laboratory strains, produced a broad range of the enzyme activity (0.5-7.9 unit/mg protein). The strains of both Streptococcus mutans and Lactobacilli showed low levels of activity (0-0.5 unit/mg protein for S. mutans). As compared with the enzyme activities of other bacteria, a moderate range of activity was produced by the strains of Strptococcus mitis nad Strptoccus salivarius. Actinomyces strains, like those of S. sanguis, produced a varying amount of activity (0-9.8 unit/ mg protein). A possible involvement of the oral bacterial PZ-peptidase in the metabolism of human saliva proteins is discussed.

• Journal of Microbiology
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• v.39 no.4
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• pp.273-278
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• 2001

Shewanela, putrefaciene IR-1 and MR-1 were cultivated by using various combinations electron donor-acceptor, lactate-Fe(III) lactate-nitrate, pyruvate-FE(III), pyruvate-nitrate H$_2$ acetate-Fe(III) and H$_2$-acetate-nitrate. Both strains grew fermentatively on pyruvate and lactate but not on without and electron acceptor. In culture with Fe(III), both astrains grew on pyruvate and lactate but on H$_2$-acetate- CO$_2$. In cultivation with nitrate, both stains grew on pyruvate lactage and on H$_2$-acetate-CO$_2$ The growth yields of IR-1 pyruvate, pyruvate-Fe(III) and lactate-Fe(III) were about 3.4, 3.5, and 3.6(g cell/M substrate), respectively. From the growth properties of both strains on media with Fe(III) as an electron acceptor, the bacterial growth was confirmed not to be increased by addition of Fee(III) as an electron acceptor to the growth medium, which indicates a possibility that the dissimilatory reduction of Fe(III) to Fe(III) may not be coupled to free energy production.

• Journal of Microbiology and Biotechnology
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• v.15 no.2
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• pp.362-367
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• 2005

On the basis of high osmotic tolerance and xylitol production, a novel yeast strain was screened from soils of rice farming. The isolated strain HY200 was systematically characterized by using general approaches of Biolog Microlog$^{TM}$ and 18S rRNA sequence analyses, and consequently was designated as Candida tropicalis HY200. Under formulated culture conditions, relatively high xylitol yield ($77\%$) and productivity (2.57 g/l$\codt$h) were obtained, in practice, when 200 g/l of xylose was supplemented. In the utilization of nitrogen, inorganic compounds could not serve as nitrogen sources. As a promising phenotype, HY200 steadily flocculated during and/or after growing in the formulated medium. The extent of flocculation was partly affected by nitrogen sources. However, regardless of the kinds of carbon source fed, the flocculent cells were always observed at the end of the exponential growth phase. These observations strongly suggest that the strain HY200 could effectively be used as a potential candidate for the production of xylitol from xylose, especially in repeated batch mode, because of its flocculation ability and tolerance to high substrate concentrations.

• The Korean Journal of Pharmacology
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• v.32 no.2
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• pp.211-219
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• 1996

The inhibitory effect of cyclosporin A (CsA) on nitric oxide production is not related to the immunosuppressive action of the drug, but to the renal toxicity and arterial hyper-tension. In this study the experimental interventions to reverse the inhibition of nitric oxide production by cyclosporin A in rat aortic smooth muscle cells were examined. CsA inhibited the accumulation of nitrite, the stable end product of nitric oxide, in culture media in a concentration $(0.1{\sim}100{\mu}g/ml)-dependent$ manner. The inhibitory effect of CsA on nitrite accumulation were not antagonized by arginine (10 mM), a substrate of nitric oxide synthase, nor by calcium ionophore A23187 $(7{\mu}M)$. Forskolin, an activator of adenylate cyclase, which enhanced iNOS induction at transcriptional level, completely reversed the inhibitory action of CsA on nitrite accumulation. However, PMA (2 nM) and PDB (50 nM), PKC activators, increased the inhibitory action of CsA on nitrite accumulalion. From these results, it is suggested that cyclic AMP-elevating agents may be candidates of therapeutic agents in prevention and treatment of renal toxicity and arterial hypertension induced by CsA. Among conventional antihypertensive drugs, calcium channel blockers and ${\alpha}-blockers$ are preferred to ${\beta}-blockers$.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.2
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• pp.105-111
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• 2002

The biological removal of 2,4,6-trinitrotoluene (TNT) was studied in a bench-scale bioreactor using a bacterial culture of strain OK-5 originally Isolated from soil samples contaminated with TNT. The TNT was completely removed within 4 days of incubation in a 2.5 L bench-scale bioreactor containing a newly developed medium. The TNT was catabolized in the presence of different supplemented carbons. Only minimal growth was observed in the killed controls and cultures that only received TNT during the incubation period. This catabolism was affected by the concentration ratio of the substrate to the biomass. The addition of various nitrogen sources produced a delayed effect for the TNT degradation. Tween 80 enhanced the degradation of TNT under these conditions. Two metabolic intermediates were detected and identified as 2-amino-4, 6-dinitrotoluene and 4-amino-2, 6-dinitrotoluene based on HPLC and GC-MS analyses, respectively. Strain OK-5 was characterized using the BIOLOG system and fatty acid profile produced by a microbial identification system equipped with a Hewlett Packard HP 5890 II gas chromatograph. As such, the bacterium was identified as a Stenotrophomonas species and designated as Stenotrophomonas sp. OK-5.