• Microbiology and Biotechnology Letters
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• v.20 no.5
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• pp.519-526
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• 1992

The gene coding for urease of alkalophilic Bacillus pasteurii had been cloned in Escherichia coli previously. The urease protein was purified 63.1-fold by TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150 and Sephadex G-200 chromatographies with a 7.3% yield from the sonicated fluid of the E. coli HB1Ol(pBUll) encoding B. pasteurii urease gene. The ureases of E. coli (pBUll) and B. pasteurii possessed as a $K_m$ for urea, 42.1 mM and 40.4 mM, respectively. They hydrolyzed urea with $V_{max}$ of 86.9$\mu$mol/min and 160$\mu$mol/min, respectively. Both ureases were composed with four subunits (Mrs 67,000) and a subunit (Mr 20,000). The molecular weight of both native enzymes was Mr 280,OOO$pm$10,000 determined by gel filtration chromatography and Coomassie blue staining of the subunits. The optimal reaction pH of both ureases were pH 7.5. The ureases were stabled in pH 5.5-10.5. The optimal reaction temperature of both ureases were $60^{\circ}C$, and the ureases were stable for an hour at $50^{\circ}C$, 40min at $60^{\circ}C$ and 10 min at $70^{\circ}C$ The activity of both enzymes were inhibited completely by $Ag^{2+}$, $Hg^{2+}$, $Zn^{2+}$, $Cu^{2+}$, and were inhibited 60% by CoH, 30% by $Fe^{2+}$ and 10% by $Pb^{2+}$. However it was increased by the addition of $Sn^{2+}$, $Mn^{2+}$, $Mg^{2+}$ at concentration of $1{\times}10^{-3}$M. Both ureases were inhibited completely by p-CMB and acetohydroxamic acid. The urease expressed in E. coli (pBU11) was inhibited 70% by SDS. The urease of B. pasteurii was inhibited 40% by hydroxyurea, whereas the recombinant urease of E. coli strain was inhibited 17%. Both enzymes were not inhibited by cyclohexanediaminetetraacetic acid (CDTA) and ethylendiaminetetraacetic acid (EDTA).

• Journal of Life Science
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• v.24 no.4
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• pp.343-351
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• 2014

Phosphoinositide 3-kinase (PI3K) plays a central role in cell signaling and leads to cell proliferation, survival, motility, exocytosis, and cytoskeletal rearrangements, as well as specialized cell responses, superoxide production, and cardiac myocyte growth. PI3K is divided into three classes; type I PI3K is preferentially expressed in leukocytes and activated by ${\beta}{\gamma}$ subunits of heterotrimeric G-proteins. In this study, the cDNAs encoding the $PI3K{\gamma}$ gene were isolated from a brain cDNA library constructed using the flounder (Paralichthys olivaceus). The sequence of the isolated $PI3K{\gamma}$ was 1341 bp, encoding 447 amino acids. The nucleotide sequence of the $PI3K{\gamma}$ gene was analyzed with that of other species, including Oreochromis niloticus and Danio rerio, and it turned out to be well conserved during evolution. The $PI3K{\gamma}$ gene was subcloned into the expression vector pET-44a(+), and expressed in the E. coli BL21 (DE3) codon plus cell. The resulting protein was expressed as a fusion protein of approximately 49 kDa containing a C-terminal six-histidine extension that was derived from the expression vector. The expressed protein was purified to homogeneity by His-tag affinity chromatography and showed enzymatic activity corresponding to $PI3K{\gamma}$. The binding of wortmannin to $PI3K{\gamma}$, as detected by anti-wortmannin antisera, closely followed the inhibition of the kinase activities. The results obtained from this study will provide a wider base of knowledge on the primary structure and characterization of the $PI3K{\gamma}$ at the molecular level.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.2
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• pp.304-314
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• 2005

The major concept of Sasang typology is that the disease susceptibility and drug response as well as physiological characteristics are presumed to be different depending on their Sasang types. Although characterizing fundamental basis of their traits are crucial in this research field, only pathological susceptibility and physical appearances were thoroughly studied. We evaluated their physiological characteristics by tapping psychological, physical and genetic traits of each Sasang types. After determining the Sasang type of one hundred three college students based on the Questionnaire for the Sasang Constitution Classification, the psychological, physical and genetic traits of each type were analyzed with the Myers-Briggs Type Indicator (MBTI), Bioelectrical Impedance Analysis and genetic polymorphism test, respectively. Each of the Sasang types showed significantly different profiles (Generalized estimation equation, coef=11.88, z=2.13, p=0.033), and could be distinctively classified based on their MBTI scores (discriminant analysis Wilks Lambda=0.611, df=8, chi-square=36.7, p<0.001). Subjects with the So-Eum type (Introversion and Judging) and the So-Yang type (Extroversion and Perceiving) showed contrasting psychological features, however they had similar anthropometric characteristics. Subjects with the Tae-Eum type showed bigger Body Mass Index ($R^2$=0.22, df=4, 74, F=5.07, p=0.001) and body shape compared to others. Although there were no significant differences in G-protein beta-3 subunit polymorphism, angiotensin-converting enzyme polymorphism and Methylenetetrahydrofolate reductase polymprhisms among groups with Sasang types, it was shown that the dopamine system could be one for genetic marker for Sasang typology. These results demonstrated distinctive and essential traits of Sasang typology using reproducible psychometric, anthropometric and genetic evaluations. We also found that the Sasang typology was a bio-psychological typology which could show trait-specific guideline for individualized medicine.