• Microbiology and Biotechnology Letters
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• v.37 no.2
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• pp.99-104
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• 2009

The $\beta$-glucosidase gene from Streptomyces coelicolor A3(2) was cloned and expressed in Escherichia coli. The ORF consisted of 1377 nucleotides encoding 51 kDa in a predicted molecular weight. Effects of pH indicated that the $\beta$-glucosidase showed similar activity using $\alpha$-pNPG($\rho$-nitrophenyl-$\alpha$-D-glucopyranoside), $\beta$-pNPG($\rho$-nitrophenyl-$\beta$-D-glucopyranoside), and $\beta$-pNPF($\rho$-nitrophenyl-$\beta$-D-fucopyranoside) at range of pH 3 to 10, and high activity using $\beta$-pNPGA ($\rho$-nitrophenyl-$\beta$-D-galactopyranoside) from pH 5 to 10, especially, 3.3 times higher activity at pH 9. Effects of temperature indicated that the $\beta$-glucosidase showed low activity using $\alpha$-pNPG, $\beta$-pNPG, and $\beta$-pNPF from $20^{\circ}C$ to $70^{\circ}C$, and increased activity using $\beta$-pNPGA from $30^{\circ}C$ to $50^{\circ}C$, 1.8 times higher activity at $50^{\circ}C$ than at $30^{\circ}C$. According to activity determination of other substrates, the enzyme was active on daidzin, genistin, and glycitin, inactive on esculin and apigenin-7-glucose. The EDTA and DTT as reducing agents inhibited $\beta$-glucosidase activity, but SDS and mercaptoethanol did not inhibit. Monovalent or divalent metal ions such as $MnSO_4$, $CaCl_2$, KCl, and $MgSO_4$ did not inhibited $\beta$-glucosidase activity. $CuSO_4$ and NaCl showed low inhibition, and $ZnSO_4$ inhibited 3.3 times higher than control.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.54 no.4
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• pp.375-383
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• 2009

This study was carried out to develop transgenic rice cultivars with the CAX1 (accession no. U57411) gene. We successfully selected the transgenic rice plants over-expressing the Arabidopsis H+/$Ca^{2+}$ antiporter CAX1 (accession no. U57411) gene in T6 generation. The brown rice of the CAX1 expressing rice contained 13.4~68.0 % more calcium $(Ca^{2+})$ than that of the wild type and 5 lines were selected based on the phenotypes compared to the control cultivar at the GMO field. The CAX1 expressing transgenic rice plants were similar in phenotype to the wild type during the whole growth period. Also these selected 4 lines appeared to be resistant to blast, cold and water solution compared with the wild type. Difference in 1,000 grain weight of brown rice was observed among each line but grain shape did not show any morphological alternations. These results suggest the enhanced Ca-substrate specificity of CAX1 exchanger in donor plant. Therefore, intact CAX1 exchanger can be functionally useful for $Ca^{2+}$ nutrient enrichment of rice with reduced accumulation of undesirable cation.

• Journal of Animal Science and Technology
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• v.46 no.1
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• pp.7-14
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• 2004

The Stearoyl-CoA Desaturase(SCD) is a key enzyme, which converting palmitic acid(16:0) and stearic acid (18:0) to pahnitoleic acid(16:1) and oleic acid(l8:1), respectively. The concentration of oleic acid(18:1) in meat of beef cattle could influence both palatability and perception of meat. This experiment has conducted to determine relationship between the compositions of monounsaturated fatty acids and the SCD mRNA level in bovine liver and loin muscle tissue. The compositions of palmitoleic acid(16:1) and oleic acid(18:1) in loin muscle were 5% and 46% of total lipid and in liver were 2% and 20% of total lipid, respectively. On the other hand, the compositions of palmitic acid(16:0) and stearic acid(18:0) in loin muscle were 25% and 45% of total lipid and in liver were 14% and 43% of total lipid, respectively. The ratio of monounsaturated to saturated fatty acids(the desaturation index) was used as a measure of SCD activity in tissues. The average desaturation index in loin muscle was higher about 3.6-fold than that in liver. The desaturation index of oleate/stearate and palmitoleatelpalmitate in loin muscle were higher 8-fold and 1.8-fold than those in liver, respectively, showing that the substrate specificity of SCD enzyme was very different between liver and muscle tissues. To determine whether the composition of monounsaturated fatty acids in liver and muscle are dependent on SCD expression, SCD mRNA level was examined by RT-PCR analysis. The SCD mRNA level in loin muscle was higher about 3-fold than that in liver. Thus, the quantitative relationship between the desaturation index of fatty acid and SCD mRNA was observed in liver and muscle. The difference in the compositions of monounsaturated fatty acids between bovine liver and muscle tissues may be due to different level of Stearoyl-CoA Desaturase mRNA.

• Microbiology and Biotechnology Letters
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• v.45 no.1
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• pp.63-70
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• 2017

A functional screen of 60,672 fosmid metagenomic clones amplified from marine sediment obtained from the Dokdo islets in Korea identified the gene EstES1, whose product, EstES1, displayed lipolytic properties on tributyrin-supplemented media. EstES1 is a 576 amino acid protein with a predicted molecular weight of 59.4 kDa including 37 N-terminal leader amino acids. EstES1 exhibited the highest sequence similarity (44%) to a carboxylesterase found in Haliangium ochraceum DSM14365. Phylogenetic analysis indicated that EstES1 belongs to a currently uncharacterized family of lipases. Within the conserved domain, EstES1 retains the catalytic triad that consists of the consensus penta-peptide motif, GESAG. EstES1 demonstrated a broad substrate specificity toward the long acyl group of ethyl esters (C2-C12), and its optimal activity was recorded toward p-Nitrophenyl butyrate (C4) at pH 9.0 and $40^{\circ}C$ (specific activity of 255.4 U/mg). The enzyme remained stable in the ranges of $60-65^{\circ}C$ and pH 9.0-10.5 and in the presence of methanol, ethanol, isopropanol, and dimethyl sulfoxide. Therefore, EstES1 has potential for use in industrial applications involving high temperature, organic solvents, and/or alkaline conditions.

• Current Research on Agriculture and Life Sciences
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• v.4
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• pp.55-64
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• 1986

Polyphenol oxidase in japanese pear (Pyrus communis var. mansamkil) was isolated, partially purified and its some properties were investigated. Polyacrylamide disc gel electrophoresis indicated two bands with polyphenol oxidase activity in the extract from acetone dry powder of par flesh. These two polyphenol oxidases (PPO A and PPO B) were purified through acetone precipitation and diethylaminoethyl cellulose column chromatography. PPO A and B were purified 7.8 fold and 8.7 fold by the present procedure, respectively. The Rm values of partially purified PPO A and B were estimated to be 0.58 and 0.68, respectively. The optimum temp, and pH of PPO A activity were $33^{\circ}C$ and pH 7.0, while those of PPO B were $30^{\circ}C$ and pH 4.2, respectively. Two PPO were unstable over the temperature of $60^{\circ}C$. The substrate specificity of pear PPO showed high affinity toward o-diphenolic compounds, especially catechol in PPO A and chlorogenic acid in PPO B, but inactive toward m-diphenol, p-diphenol and monophenols. PPO A showed affinity toward the trihydroxyphenolic compound. $Zn^{{+}{+}}$ activated the PPO A activity but $Fe^{{+}{+}}$ inhibited PPO B activity, while $Fe^{{+}{+}}$ and $Zn^{{+}{+}}$ activated the PPO B activity, while $Fe^{{+}{+}}$ and $Zn^{{+}{+}}$ activated the PPO B activity but $K^+$, $Mg^{{+}{+}}$, $Ca^{{+}{+}}$ and $Hg^{{+}{+}}$ inhibited at 10mM concentration. $Cu^{{+}{+}}$ activated the enzyme action at low concentrations but inhibited at high concentration. Inhibition studies indicated that L-ascorbic acid, L-cysteine and thiourea were most potent. The Km values of PPO A and PPO B for catechol were 20mM and 14.3mM, respectively.