• Journal of Microbiology and Biotechnology
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• v.9 no.6
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• pp.804-810
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• 1999

There is a possibility that carvone, a monoterpene from spearmint (Mentha spicata), could induce the bph degradative pathway and genes in Alcaligenes eutrophus H850, which is a known Gram-negative PCB degrader with a broad substrate specificity that was thoroughly investigated with Arthrobacter sp. BIB, a Gram-positive PCB degrader. The strains BIB and H850 were unable to utilize and grow on the plant terpene [(R)-(-)-carvone] (50ppm) to be recognized as a sole carbon source. Nevertheless, the carvone did induce 2,3-dihydroxybiphenyl 1,2-dioxygenase (encoded by bphC) in the strain B lB, as observed by a resting cell assay that monitors accumulation of a yellow meta ring fission product from 4,4'-dichlorobiphenyl (DCBp). The monoterpene, however, did not appear to induce the meta cleavage pathway in the strain H850. Instead, an assumption was made that the strain might be using an alternative pathway, probably the ortho-cleavage pathway. A reverse transcription (RT)-PCR system, utilizing primers designed from a conserved region of the bphC gene of Arthrobacter sp. M5, was employed to verify the occurrence of the alternative pathway. A successful amplification (182bp) of mRNA transcribed from the N-terminal region of the bphC gene was accomplished in H850 cells induced by carvone (50ppm) as well as in biphenyl-growth cells. It is, therefore, likely that H850 possesses a specific PCB degradation pathway and hence a different substrate specificity compared with B1B. This study will contribute to an elucidation of the dynamic aspects of PCB bioremediation in terms of roles played by PCB degraders and plant terpenes as natural inducer substrates that are ubiquitous and environmentally compatible.

• Journal of Microbiology and Biotechnology
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• v.31 no.3
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• pp.464-474
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• 2021

Bacterial cytochrome P450 (CYP) enzymes are responsible for the hydroxylation of diverse endogenous substances with a heme molecule used as a cofactor. This study characterized two CYP154C3 proteins from Streptomyces sp. W2061 (CYP154C3-1) and Streptomyces sp. KCCM40643 (CYP154C3-2). The enzymatic activity assays of both CYPs conducted using heterologous redox partners' putidaredoxin and putidaredoxin reductase showed substrate flexibility with different steroids and exhibited interesting product formation patterns. The enzymatic characterization revealed good activity over a pH range of 7.0 to 7.8 and the optimal temperature range for activity was 30 to 37℃. The major product was the C16-hydroxylated product and the kinetic profiles and patterns of the generated hydroxylated products differed between the two enzymes. Both enzymes showed a higher affinity toward progesterone, with CYP154C3-1 demonstrating slightly higher activity than CYP154C3-2 for most of the substrates. Oxidizing agents (diacetoxyiodo) benzene (PIDA) and hydrogen peroxide (H2O2) were also utilized to actively support the redox reactions, with optimum conversion achieved at concentrations of 3 mM and 65 mM, respectively. The oxidizing agents affected the product distribution, influencing the type and selectivity of the CYP-catalyzed reaction. Additionally, CYP154C3s also catalyzed the C-C bond cleavage of steroids. Therefore, CYP154C3s may be a good candidate for the production of modified steroids for various biological uses.

• Microbiology and Biotechnology Letters
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• v.18 no.5
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• pp.484-489
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• 1990

An intracellular inulase ( fJ-fructofuranoside fructohydrolase, EC 3.2.1.26) from Bacillus subtilis has been partially purified and its mode of action and general properties were studied. The enzyme had an apparent molecular weight of 49,000 as estimated by gel filtration and its pI point was 5.2. Substrate concentration studies showed an apparent Km of 10 mM for sucrose and of 18 mM for raffinose. The enzyme was an acid-labile protein with a pH optimum of 6.6. The optimum temperature was 50$^{\circ}$C. The enzyme acts on straight chain oligo- and poly-fructosides of the inulin series via a exo-wise cleavage mechanism, as well as on sucrose.

• Korean Journal of Animal Reproduction
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• v.12 no.2
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• pp.120-124
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• 1988

Rat oocyte-cumulus complexes were cultured in verious media in order to induce maturation division in vitro. When the complexes were cultured in mKRB containing estrous rat serum (ERS) and FCS of 5% instead of BSA higher proportions(83.3 and 86.7%) of oocytes matured to metaphase II in 20h compared to control(75%) and t도 maturation rates in mKRB plus FCS were generally higher than those in mKRB plus ERS. Fertilization and early cleavage rates in vitro of the intact oocytes matured in mKRB containing BSA and FCS were generally higher than those of cumulus-removed oocytes and these rates were higher in mKRB containing 5% FCS than those in mKRB containing BSA. These results indicate that maturation rate in vitro was greatly increased by the addition of FCS instead of BSA to mKRB solution and the presence of cumulus cells around oocytes prior to sperm insemintion may be responsible for the increase of in vitro fertilization and early cleavage rates.

• Microbiology and Biotechnology Letters
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• v.24 no.6
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• pp.743-748
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• 1996

For the biological treatment of phenolic resin wastewater containing phenol and formaldehyde, a phenol-degrading yeast was isolated from the papermill sludge, and then identified as Candida tropicalis PW-51 according to morphological, physiological and biochemical properties. The strain was able to degrade high phenol concentrations up to 2,000mg/l within 58 hours in batch cultures. Phenol-degrading efficiency by the strain was maximum at the culture conditions of a final concentration of 9 $\times$ 10$^{6}$ cells/ml, 30$\circ$C and pH 7.0. The mean degradation rate of phenol was highest at 45.5mg/l/h in 1,000mg/l phenol from 500mg/l to 2,000mg/l phenol. Because the enzyme activity of catechol 1,2-dioxygenase increased in the course of degradation of phenol, it seems that this strain degrades phenol via the ortho-cleavage of benzene ring. The isolate C. tropicalis PW-51 could be effectively used for the biological treatment of phenolic resin wastewater.

• Microbiology and Biotechnology Letters
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• v.16 no.3
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• pp.199-204
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• 1988

A strain capable of growth on naphthalene minimal medium was isolated from soil by selective enrichment culture and identified as Pseudomonas putida N3 according to its morphological and physiological characteristics. The optimum pH and temperature for growth of the isolate were 7.0 and 3$0^{\circ}C$, respectively. This strain was resistant to ampicillin, chloramphenicol, kanamycin and streptomycin but. sensitive to tetracycline and rifampicin. Of the naphthalene related compounds, 1, 5-dihydroxynaphthalene was more easily utilized than naphthalene due to its solubility. And catechol was degraded through meta-cleavage pathway. A 110 Kb plasmid which encodes for a single set of enzymes responsible for the degradation of naphthalene was obtained.

• Journal of Embryo Transfer
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• v.22 no.2
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• pp.121-126
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• 2007

The objective of this study was carried out to examine the selection effects of in vitro matured porcine follicular oocytes with polar body extrusion and early cleavage as non-invasive marker to know the developmental competence in advance. The porcine oocytes matured for 48 h were examined the polar body extrusion. The examined oocytes were matured for additional $16{\sim}18h$ and activated with 7% ethanol and cultured in $5{\mu}g/ml$ cytochalasin B for 5 h for diploid formation. The treated oocytes were cultured and examined the cleavage after 48 h and continued culturing for 5 days. The oocytes of 21.9% were discarded in morphological selection and 32.1% oocytes were discarded by failure of first polar body extrusion. The selected oocytes were matured and activated and then after 48 h the cleavage rates were examined. In morphologically selected oocytes, 15.8% oocytes were not cleaved and 52.6% oocytes were normally cleaved and 31.6% oocytes were hyper-cleaved over 8-cell stage. However in the first polar body extruded oocytes, 7.1% oocytes were not cleaved and 73.1% oocytes were normally cleaved and 19.8% oocytes were hyper-cleaved. The morphologically selected embryos that not cleavage-selected were developed in 16.7% up to blastocyst and the morphologically selected and cleavage-selected embryos were developed in 31.7%. The polar body extruded oocytes that were not carried out cleavage selection were developed in 39.0% and the polar body extruded and cleavage-selected embryos were developed 49.0%. The first cleavage timing was examined with 12 h interval after activation. In $0{\sim}12,\;12{\sim}24,\;24{\sim}36,\;and\;36{\sim}48h$ intervals, 4.1%, 68.6%, 19.1%, and 2.3% oocytes were cleaved and 5.9% oocytes were not cleaved until 48 after activation. The cleaved oocytes in each interval were cultured and developed upto blastocyst with 0, 39.1, 9.5, and 0%, respectively. This results suggests that polar body extruded and cleaved at $12{\sim}36h$ embryo has higher developmental potential than the others.

• Bulletin of the Korean Chemical Society
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• v.6 no.6
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• pp.362-366
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• 1985

The hydrogen atom transfer reaction between substituted methane, $CH_3X,$ and its radical, $CH_2X(X=H,F,CH_3,CN,OH\;and\;NH_2$ was studied by MINDO/3 method. The transition state(TS) structure and energy barriers were determined and variation of the transition state and of the reactivity due to the change of X were analyzed based on the potential energy surface characteristics. It was found that the greater the radical stabilization energy. the looser the TS becomes; the TS occurs at about 15% stretch of the C-H bond, which becomes longer as the radical stabilization energy of $CH_2X$ increasers. The intrinsic barrier, ${\Delta}E*_{x.x},$ of the reaction with X was found to increase in the order $H The degree of bond stretch of the C-H bond stretch of the C-H bond at the TS also had the same order indicating that the homolytic bond cleavage of the C-H bond is rate-determining. Orbital interactions at the TS between LUMO of the fragment $C{\ldots}H{\ldots}C$ and the symmetry adapted pair of nonbonding, $n{\pm}(=n_1{\pm}n_2),$ or pi orbitals of the two X atoms were shown to be the dominant contribution in determining tightness or looseness of the TS. The Marcus equation was shown to apply to the MINDO/3 barriers and energy changes of the reaction.

• Bulletin of the Korean Chemical Society
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• v.28 no.6
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• pp.917-920
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• 2007

Ketene-forming eliminations from C4H3(S)CH2C(O)O-C6H3-2-X-4-NO2 (1) promoted by R2NH in MeCN have been studied kinetically. The reactions are second-order and exhibit Bronsted β =0.51-0.62 and |βlg|= 0.47-0.53. Hence, an E2 mechanism is evident. The Bronsted β increased from 0.33 to 0.53 and |βlg| remained nearly the same by the change of the base-solvent from Bz(i-Pr)NH/Bz(i-Pr)NH2+ in 70 mol% MeCN(aq) to Bz(i-Pr)NH-MeCN, indicating a change to a more symmetrical transition state with similar extents of Cβ -H and Cα -OAr bond cleavage. When the β-aryl group was changed from thienyl to phenyl in MeCN, the β value increased from 0.53 to 0.73 and |βlg| decreased from 0.53 to 0.43. This indicates that the transition state became skewed toward more Cβ -H bond breaking with less Cα-OAr bond cleavage. Noteworthy is the greater double bond stabilizing ability of the thienyl group in the ketene-forming transition state.

• 대한생식의학회:학술대회논문집
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• 2003.12a
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• pp.122.2-123
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• 2003