• Journal of Life Science
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• v.15 no.3 s.70
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• pp.415-422
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• 2005

library of the mutants was prepared by transposon mutagenesis of the Mycobacterium bovis BCG. We screened this library for the resistance to an anti-tuberculosis antibiotic, PA-824. Most of the mutants resistant to the PA-824 were not able to synthesize the coenzyme $F_{420}$ which is normally produced by the wild type M. bovis BCG strains. HPLC analysis of the cellular extract showed that one of those mutants which lost the ability to synthesize $F_{420}$ still produced F0. The insertion site of the transposon in this mutant was determined by an inverse PCR and the transposon was found to be inserted in the Rv2435c open reading frame (ORF). Rv2435c ORF is predicted to encode an 80.3 kDa protein. Rv2435c protein appears to be bound to the cytoplasmic membrane, its N-terminal present in the periplasm and C-terminal in the cytoplasm. The C-terminal portion of this protein is highly homologous with the adenylyl cyclases of both prokaryotes and eukaryotes. There are 15 ORFs which have homology with the class III AC proteins in the genome of the M. tuberculosis and M. bovis. Two of those, Rv1625c and Rv2435c, are highly homologous with the mammalian ACs. We cloned the cytoplasmic domain of the Rv2435c ORF and expressed it with six histidine residues attached on its C-terminal in Escherichia coli to find out if this protein is a genuine AC. Production of that protein in E. coli was proved by purifying the histidine-tagged protein by using the Ni-NTA resin. This protein, however, failed to complement the cya mutation in E. coli, indicating that this protein lacks the AC activity. All of the further attempts to convert this protein to a functional AC by a mutagenesis with UV or hydroxylamine, or construction of several different fusion proteins with Rv1625c failed. It is, therefore, possible that Rv2435c protein might affect the conversion of F0 to $F_{420}$ not by synthesizing cAMP but by some other way.

• Korean Journal of Food Science and Technology
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• v.42 no.1
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• pp.1-7
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• 2010

To revise the dithiocarbamates residue analysis method and survey the residues in agricultural products that were treated with these fungicides in Korea, we purchased 20 types of foodstuffs (rice, potato, cabbage, apple etc.) from markets in five major cities. 236 samples of the purchased foodstuffs were then analyzed for the presence of dithiocarbamates by HPLC/UV and HPLC/APCI-MS. The $R^2$, LOD and LOQ in the range of 0.5-107.3 mg/L were as follows: DCC: y=174.34x+18.315, $R^2=0.9999$, 0.01 mg/L, and 0.04 mg/L; EBDC: y=227.38x-14.715, $R^2=1.0000$, 0.01 mg/L and 0.02 mg/L; PBDC: y=38.46x-21.412, $R^2=0.9999$, 0.04 mg/L, and 0.1 mg/L; ETU: y=52.752x-4.4819, $R^2=0.9998$, 0.02 mg/L and 0.03 mg/L; PTU: y=128.28x+4.4624, $R^2=0.9998$, 0.02 mg/L, and 0.04 mg/L. The levels of DDC, EBDC, PBDC, ETU and PTU in 20 agricultural products fortified to 10.0-107.3 mg/L ranged from 61.7-117.5%, 65.3-110.1%, 61.5-109.6%, 69.3-116.3% and 70.2-97.2%, respectively. Overall, dithiocarbamates were detected in 100 samples and the detection ratio was 42.4%. Among these, only 3 samples (1.3%) of Lycii fructus had residue levels that were above the action limits, while the remaining samples (233 samples) contained levels of dithiocarbamates below the detection limit or below the Korea MRLs (Maximum Residue Limits).

• Korean Journal of Animal Reproduction
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• v.27 no.2
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• pp.153-161
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• 2003

This study has evaluated effect of the spermatozoa incubation on the glycosidase activity and fertilizing ability in vitro in the pig. To identify sperm glycosidases specific for sugar residues found in the zona pellucida of pig oocytes, the spermatozoa were treated experimentally and assayed for activities of $\alpha$-L-fucosidase, $\alpha$-D-mannosidase, $\beta$-D-galactosidase and N-acetyl-$\beta$-D-glucosaminidase ($\beta$-GlcNAc'ase). The glycosidases activity were higher in spermatozoa incubated for 2h than without incubation. The $\beta$-GlcNAc'ase activity was at least two-fold higher than other glycosidase regardless of spermatozoa incubation. In the same glycosidases, the activity had a tendency to increase as time of spermatozoa incubation was prolonged, but there were no differences in spermatozoa incubated during the various periods (4~24h). The percentages of spermatozoa that reached acrosome reaction were affected by glycosidases in the medium (P<0.05, for mannosidase), and were higher in spermatozoa with that than without incubation. On the other hand, the spermatozoa motility were decreased with incubation periods, but no effects by different glycosidases on the change of sperm motility during the various periods of incubation. In other experiment, the binding and penetration of pig spermatozoa were tested with oocytes matured in vitro in the presence of various glycosidase. The penetration rates were decreased with incubation of spermatozoa when oocytes were inseminated in medium with different glycosidases. These rates were higher in spermatozoa non-incubated than with incubation for 2h (P<0.05 for GlcNAc'ase; P<0.01 for control group). The sperm-zona binding rate in control group were higherthan in medium with glycosidases. In addition, the highest binding rate were obtained in medium with GlcNAc'ase. In all glycosidases, the sperm-zona binding rate in spermatozoa without incubation were higher than incubation for 2h. The significant differences were obtained in spermatozoa treated with $\alpha$-D-mannosidase (P<0.05). These results suggest that $\beta$-GlcNAc'ase is present mainly in the plasma membrane of pig spermatozoa. It was also shown that the glycosidase activity were increased in all glycosidases in spite of low sperm-zona binding rate and penetration rates by spermatozoa incubation.

• Korean Journal of Environmental Agriculture
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• v.34 no.4
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• pp.274-281
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• 2015

BACKGROUND: A lasting release of low levels of persistence chemicals including pesticides and pharmaceuticals into river has a bad influence on aquatic ecosystems and humans. The present study monitored pesticide residues in the Yeongsan and Seomjin river basins and their tributaries as a fundamental study for water quality standard of pesticides.METHODS AND RESULTS: Nine pesticides(aldicarb, carbaryl, carbofuran, chlorpyrifos, 2,4-D, MCPA, methomyl, metolachlor, and molinate) were determined from water samples using SPE-Oasis HLB(pH 2) and LC/MS/MS. Validation of the method was conducted through matrix-matched internal calibration curve, method detection limit(MDL), limit of quantification(LOQ), accuracy, precision, and recovery. MDLs of all pesticides satisfied the GV/10 values. Linearity(r²) was 0.9965- 0.9999, and a percentage of accuracy, precision, and recovery was 89.4-113.6%, 3.1-14.0%, and 90.8-106.2%, respectively. All pesticides exclusive of aldicarb were determined in the river samples, and there was a connection between the positive monitoring results and agricultural use of the pesticides.CONCLUSION: Monitoring outcomes of the present study implied that pesticides were a possible non-point pollutant source in the Yeongsan and Seomjin river basins and tributaries. Therefore, it is required to produce and accumulate more monitoring results on pesticides in river waters to set water quality standards, finally to preserve aquatic ecosystems.

• Applied Microscopy
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• v.41 no.1
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• pp.47-53
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• 2011

Hair is an appendage of skin which protects the body from outer physical and chemical stimuli. Hair is generated from the hair follicle lying on a sunken basal layer of epidermis. Hair cycling, which regenerates hair follicles throughout the life time of the organism. Numerous kinds of factors which exist at the hair follicle have been reported to regulate hair cycling, Human growth hormone secreted from pituitary gland, initially demonstrated to accelerate organ's growth, has been reported to play a role in the biology of organ size determination. We investigated the effect of 6-histidines residues tagged at amino-terminus of human growth hormone using light and electronmicroscopic methods. Human growth hormone encapsulated in nano-liposome (LhGH) was used to find how LhGH affects hair follicle cycling of mouse (C57BL6/CrN). Distilled water as a negative control, 3% Minoxidil as a positive control, and LhGH were applied to mouse for weeks. LhGH increased the number of exposed hairs per given areas ($1mm^2$). This result was also confirmed using a different breed of mice which show natural hair loss in an old age (about 17 months after birth). When LhGH was applied for 3 weeks after natural hair loss, natural hair loss on these mice was prevented, However, the control group mice on which LhGH was not applied showed further hair loss. This result indicates that LhGH may stimulate hair cycling of mouse. In clusion, it is cleat that the LhGH increased the number of hair on mice and help the depilated skin to grow new hair follicles again.

• Horticultural Science & Technology
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• v.30 no.4
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• pp.345-356
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• 2012

We investigated the changes of fruit quality parameters, polysaccharide contents and cell wall components during maturation and ripening of two Korean pear cultivar 'Hanareum' and 'Manpungbae' compared with 'Niitaka' pear (Pyrus pyrifolia Nakai) which showed different physiological maturity based on days after full bloom (DAFB). Flesh firmness decreased continuously with fruit development and maturation, reaching a final level of 29.4, 33.5, and 27.4N at maturity in 'Hanareum' (127 DAFB), 'Manpungbae' (163 DAFB), and 'Niitaka' (170 DAFB), respectively. The level of ethylene production was very low in early season 'Hanareum' pear which showed at most 0.39 ${\mu}L{\cdot}L^{-1}$ at maturity and no ethylene was detected in 'Manpungbae' and 'Niitaka' at maturity. Fructose was the most abundant soluble sugar during fruit maturation in the pears tested and an increase of sucrose was observed during fruit ripening in the Asian pears commonly. Ethanol insoluble solids (EIS) content decreased gradually with different levels among the pear cultivars as fruit ripens consisted of 10.79, 12.72, and 12.75 $mg{\cdot}g^{-1}$ FW. The amount of total soluble polyuronides was higher in early season cultivars 'Hanareum' than those of mid-season cultivar 'Manpungbae' and 'Niitaka'. In 'Niitaka' which harvested most late season, the level of 4% KOH soluble hemicelluloses was lower than 'Hanareum' and 'Manpungbae' and maintained constantly during fruit ripening period. Cellulosic residues were determined high level in 'Niitaka' which showed 612.33 ${\mu}g{\cdot}mg^{-1}$ EIS at maturity when compared with 'Hanareum' (408.0 ${\mu}g{\cdot}mg^{-1}$ EIS) and 'Manpungbae' (538.67 ${\mu}g{\cdot}mg^{-1}$ EIS). The main constituents of cell wall neutral sugars which consisted of arabinose, xylose, galactose, and glucose were decreased gradually with onset of fruit ripening regardless of cultivar. Arabinose which was predominant in 'Hanareum' pear decreased at the last stage of ripening, but the changes of cell wall neutral sugar during ripening were not occurred in 'Niitaka' pear. The change of molecular mass distribution in water soluble pectin observed dominantly at the early stage of fruit development. Depolymerization of 4% KOH-soluble hemicelluloses and degradation of xyloglucan showed in early-season cultivar 'Hanareum' during fruit maturation, and degradation of those fractions were detected only at the early stage fruit development in mid-season cultivar 'Manpungbae' and 'Niitaka'. The molecular mass profile of CDTA soluble pectin, $Na_2CO_3$-SP and 24% KOH soluble hemicelluloses showed no significant change during fruit maturation regardless of cultivar.

• Journal of Food Hygiene and Safety
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• v.27 no.3
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• pp.224-232
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• 2012

This study was performed to investigate the pesticide residue of commercial medicinal plants used for food materials in the Seoul area. Multi class pesticide multiresidue methods in Korea Food Code was used to analyze 100 pesticides. Analyzed samples were 261 cases(domestic 201, imported 60), detection rate was 19.2%(domestic 20.9%, imports 13.3%). 17 pesticides were detected in fruit(chinese matrimony vine, jujube, rubus coreanus, japanese cornlian cherry, schizandra, tangerine peel), and root(cnidium, licorice, astragalus). Pesticide over Maximum Residue Limits were detected in jujube, cnidium. Frequently detected pesticides were cypermethrin, chlorpyrifos, cyhalothrin, fenvalerate, bifenthrin. More than 50% of the sample were detected two or more pesticides at the same time. Because of the variety and increase of pesticide detection in medicinal roots and fruits, continued monitoring and safety management is required.

• Journal of Life Science
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• v.16 no.6
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• pp.1014-1028
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• 2006

This study attempted to investigate the developmental alterations of the cerebral cortex. The effects of EGEE on the developmental cerebral cortex in the prenatal, postnatal and adults were examined by lectin histochemical methods. To investigate the change of sugar residues of glycoconjugates, biotinylated lectins(DBA, SBA, PNA, BSL-1, RCA-1, sWGA, UEA-1, Con A and LCA) were detected with by IHC using ABC kit. In the case of injection of EGEE, the reactions to Con A and LCA were shown in binding phase in the cerebral cortex commonly, and the reactions to PNA, RCA-1 and LCA were shown partially, the number of lectins to be shown reaction were decreased, and there were no reactions to DBA, SBA, BSL-1, RCA-1 and UEA-1. The reaction to Con A was similar to control group during developmental phases. The reaction to LCA was increased in the fetal, suckling, and weanning phases compared with control group. But there were no reactions to SBA and sWGA, the reaction to PNA was decreased in the frontal and occipital cortex and no reaction to sWGA in the fetal phase. There were no reactions to sWGA and PNA in the suckling phase and, no reaction to PNA and sWGA. The reaction to Con A was decreased in the frontal, parietal and occipital cortexes and, the reaction to LCA was decreased in the frontal and occipital cortexes in adult phase. As the results, the effects of EGEE, environmental hormone on the each part of cerebral cortex have shown differences. But, It had deep effect on the differentiation and growth in the cerebral cortex pathologically. In particular, the effect was severe in suckling phase. $Galactosyl-({\beta}-1,3)-N-acetyl-D-galactosamine$,${\beta}-N-acetyl-D-galactosamine$ and ${\beta}-N-acetyl-D-glucosamine$ were decreased while ${\alpha}-D-mannose$ and ${\alpha}-D-glucose$ were increased. It affected the sugar metabloism, and it was severe in fetal and suckling phases.

• Journal of Life Science
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• v.23 no.1
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• pp.15-23
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• 2013

AprE51 from Bacillus amyloliquefaciens CH51 is a 27 kDa subtilisin-like protease with fibrinolytic activity. AprE51-6 showing increased catalytic activity was produced previously. To enhance the thermostability of AprE51-6, 2 residues, Gly-166 and Asn-218 based on B. subtilis subtilisin E were mutated by site-directed mutagenesis. The results of the mutational analysis showed that substitution of arginine for Gly-166 (AprE51-7) increased the fibrinolytic activity 1.8-fold. An N218S mutant (AprE51-8) also increased the fibrinolytic activity up to 4.5-fold in a fibrin plate assay. Purified AprE51-7 and AprE51-8 mutants had a 1.9- and a 2.5-fold higher $k_{cat}$, respectively, and a 2.1-1.9-fold lower $K_m$, respectively. This resulted in a 3.8- and a 4.7-fold increase in catalytic efficiency ($k_{cat}/K_m$), respectively, relative to that of wild-type AprE51. AprE51-8 had a broader pH range than AprE51-6 and nattokinase, especially at an alkaline pH value. In addition, AprE51-8 showed higher thermostability than AprE51-6 at $60^{\circ}C$. The half-lives of AprE51-7 and AprE51-8 at $50^{\circ}C$ were 21.5 and 27.3 min, respectively, which are 2.0 and 2.6 times longer, respectively, than that of the wild-type AprE51.

• Journal of Life Science
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• v.27 no.2
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• pp.139-148
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• 2017

The inhibition of tyrosinase, a key enzyme in mammalian melanin synthesis, plays an important role in preventing skin pigmentation and melanoma. Therefore, tyrosinase inhibitors are very important in the fields of medicine and cosmetics. However, only a few tyrosinase inhibitors are currently available because of their toxic effects on skin or lack of selectivity and stability. Therefore, we synthesized a novel series of (E)-2-(substituted benzylidene)-2,3-dihydro-1H-cyclopenta[a]naphthalen-1-one derivatives and evaluated their inhibitory effects on mushroom tyrosinase, with the aim of discovering a novel tyrosinase inhibitor. Among 19 derivatives, MHY3655 ($IC_{50}=0.1456{\mu}M$) showed the strongest inhibitory effect on tyrosinase activity compared to kojic acid ($IC_{50}=17.2{\mu}M$), a well-known tyrosinase inhibitor. In addition, MHY3655 showed competitive inhibition on Lineweaver-Burk plots. We confirmed that MHY3655 strongly interacts with mushroom tyrosinase residues through the docking simulation. Substitutions with a hydroxy group at both R2 and R4 in the phenyl ring indicated that these groups play a major role in the high binding affinity to tyrosinase. Further, MHY3655 did not show cytotoxicity at the concentrations tested in B16F10 melanoma cells. In conclusion, the novel compound MHY3655 potentially shows tyrosinase inhibitory activity, and it could be used as an ingredient in whitening cosmetics.