• BMB Reports
• /
• v.33 no.4
• /
• pp.353-357
• /
• 2000

A new assay method of ${\alpha}-Oxidase$ (fatty acid : oxygen dioxygenase, 1-decarboxylating) was developed using a bioluminescence reaction system of marine luminous bacterium, Photobacterium phosphoreum. ${\alpha}$-Oxidase was isolated from a cucumber (Cucumis sativus). Pentadecanoic acid was used as a substrate, and the product, tetradecanal, was analyzed with a bacterial luciferase-coupled reaction. Initial light intensity was directly related to the concentration of tetradecanal in the range of 1 nM to 10 ${\mu}M$. Optimal pH and temperature were 7.5 and $25^{\circ}C$, respectively. Optimal pentadecanoic acid concentration in a standard assay of ${\alpha}$-oxidase was 0.1 mM. The Km value of pentedecanoic acid was $85{\mu}M$. This method is straightforward, rapid, convenient, and easy. Its needs no treatment or extraction of reaction mixture.

• BMB Reports
• /
• v.29 no.4
• /
• pp.300-307
• /
• 1996

ATP and ADP are potential regulators of mitochondtial respiration and at physiological concentrations they affect the rate of electron transfer between cytochrome c and cytochrome c oxidase. The electron transfer, however, depends on the electrostatic interaction between the two proteins. In order to exclude any nonspecific ionic effects by these polyvalent nucleotides, we used 2'-O-(2,4,6)trinitro(TNP)-derivatives of ATP and ADP which have three orders of magnitude higher affinity for cytochrome c oxidase. A simple titration of the fluorescence intensity of TNP by cytochrome c oxidase showed a binding stoichiometry of 2:1 cytochrome c:cytochrome c oxidase. Higher ionic strength was required for TNP-ATP than for TNP-ADP to be dissociated from cytochrome c oxidase, indicating that the negative charges on the phosphate group are at least partially responsible for the binding. In both spectrophotometric and polarographic assays, addition of ATP (and ADP to a less extent) showed an enhanced cytochrome c oxidase activity. Both electron paramagnetic resonance and fluorescence spectra indicate that there is no Significant change in the cytochrome c-cytochrome c oxidase interaction. Instead, reduction levels of the cytochromes at steadystate suggest that the increased activity of nucleotide-bound cytochrome c oxidase is due to faster electron transfer from cytochrome ${\alpha}$ to cytochrome ${\alpha}_3$, which is known to be the fate limiting step in the oxygen reduction by cytochrome c oxidase.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.24 no.6
• /
• pp.843-847
• /
• 1995

To evaluate the effect of dietary fiber on the protein utilization, Sprague-Dawley rats were fed diet containing 15% or 30% of pectin and 15% or 30% ${\alpha}-cellulose$. Control group was fed fiber free diet. The animals were fed and libitum for 5 weeks. Weight gain was less in the rat fed a pectin supplemented diet than those fed ${\alpha}-cellulose$ supplemented or control diet. Furthermore, weight gain decreased more by the addition of 30% pectin than 15% pectin level. The rats fed ${\alpha}-cellulose$ or pectin shwoed a decreasing tendency of food efficiency ratio compared to the control group. The rats fed a diet containing pectin showed an increasing tendency of the liver weight compared to the control group and those fed cellulose. The rats fed a diet containing pectin showed a decreasing tendency of hepatic protein content compared to those fed cellulose or control group fed fiber free diet. The rats fed diet containing pectin(15%, 30%) showed remarkable decreased activity of liver xanthine oxidase compared with those fed ${\alpha}-cellulose$ or the control group. These results suggested that the pectin may be alter the absorption of protein in intestinal lumen.

• Applied Biological Chemistry
• /
• v.14 no.2
• /
• pp.131-135
• /
• 1971

1. L-Tyrosine-${\alpha}$-ketoglutaric transaminase and p-hydroxyphenylpyruvic acid oxidase are distributed in Aspergillus oryzae. 2. L-Tyrosine oxidation in extracts of acetone powder, cell free extract and culture liquid of Aspergillus oryzae cultivated in the shaking culture are considerably accelerated by the addition of ${\alpha}$-ketoglutaric acid and then formation of glutamic acid was identified by chromatography method. 3. The roles of ${\alpha}$-ketoglutaric acid and pyridoxal phosphate have been shown to be an amino group acceptor in a transamination reaction. 4. Enzyme systems of an extracts of acetone powder and cell free extract also rapidly oxidized L-tyrosine and p-hydroxyphenlpyruvic acid to homogentisic acid. 5. The optimum pH for L-tyrosine-${\alpha}$-ketoglutaric acid transaminase was pH values of 6.0 and 6.5, and that for p-hydroxyphenylpyruvic acid oxidase was at pH values of 7.5.

• Korean Journal of Microbiology
• /
• v.40 no.3
• /
• pp.248-253
• /
• 2004

To scrutinize the physiological diversity by BIOLOG microplate, the carbon source utilization patterns of 168 strains of oligotrophic bacteria and 132 strains of psychrotrophic bacteria isolated from Lake Baikal during 2000 and 2002 were investigated. Eighty-six percent (56 strains) of oxidase test positive group (GN-NENT group) and 89 % (92 strains) of oxidase test negative group (GN-ENT group) among oligotrophic bacteria, and 82% (85 strains) of oxidase test negative group among psychrotrophic bacteria were able to utilize $\alpha$-D-glucose as a sole-carbon-source, and 93% (26 strains) of oxidase test positive group among psychrotrophic bacteria were able to utilize bromosuccinic acid as a sole-carbon-source. However, most strains except few oligotrophic bacteria with oxidase test negative group were not able to utilize $\alpha$-D-lactose as a sole-carbon-source. Most dominant genus among 300 strains was Pseudomonas (49 strains). Other dominant genera belonged to Salmonella, Serratia, Buttiauxella, Pantoea, Yersinia, Brevundimonas, Hydrogenophaga, Photorhabdus, Sphingomonas, and Xenorhabdus. Our results by BIOLOG identification system were able to provide basic data to determine community-level carbon source utilization patterns and to accomplish the efficient and reliable identification for microbial community structure in Lake Baikal.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.45 no.9
• /
• pp.1385-1390
• /
• 2016

Gallic acid is a representative hydroxybenzoic acid and is found in free form in several plants and in various esterified forms as a part of hydolyzable tannins. Convenient enzymatic transformation of trihydroxylated gallic acid with polyphenol oxidase originating from pear was evaluated to investigate whether polyphenol oxidase can be used as a valuable compound to improve the biological activity of gallic acid. Enzymatic oxidation processing of gallic acid using polyphenol oxidase was carried out for five different reaction times. The antioxidant effects of transformed gallic acid for different reaction times were evaluated via radical scavenging assays using 1,1-diphenyl-2-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radicals. In addition, the anti-diabetic property of the transformed gallic acid was measured based on ${\alpha}$-glucosidase. Gallic acid reacted for 5 h showed significantly higher antioxidant and ${\alpha}$-glucosidase inhibitory activities compared to the tested positive control substances. Biotransformation of simple gallic acid induced by polyphenol oxidase might be responsible for enhancing the biological activity of gallic acid.

• Microbiology and Biotechnology Letters
• /
• v.26 no.1
• /
• pp.68-75
• /
• 1998

The overproduction and high level secretion of Glucose Oxidase (GOD) from A. niger in S. cerevisiae was carried out by cloning GOD gene. For this purpose, using two different strong promoters (ADH1 promoter, GAL10 promoter) and signal sequences (${alpha}$-MF signal sequence of S. cerevisiae and ${alpha}$-amylase signal sequence of A. oryzae) and GAL7- and GOD terminator, four expression vectors were constructed. All the expression vectors were transformed in S. cerevisiae 2805 using auxotroph method. By the flask culture, transformants of pGAL expression vector series containing GAL 10 promotor showed much higher GOD productivity than transformants of pADH expression vector series containing ADH1 promoter Transformants of pGALGO2 containing GAL10 promotor and ${alpha}$-amylase signal sequence has shown the best productivity of GOD ($GOD_{total}$: 10.3 unit/mL, $GOD_{ex}$: 8.7 unit/mL) at 115 hr. This value was three fold higher than that of pGALGO1 containing GAL 10 promotor and ${alpha}$-MF signal sequence, even if the same promotor was involved. Through the ${alpha}$-amylase signal sequence of A. oryzae, GOD was secreted much more than the case of ${alpha}$-MF signal sequence from S. cerevisiae. These results suggest that signal sequence may play a important roles in not only the secretion but also the overproduction of foreign protein. Secretion rate of GOD in pGALGO1 and pGALGO2 was 89% and 84%, respectively, Because of the overglycosylation in S. cerevisiae the molecular weight of recombinant GOD in S. cerevisiae was much larger (250 kDa) than that of nature GOD in A. niger (170 kDa).

• Preventive Nutrition and Food Science
• /
• v.7 no.1
• /
• pp.18-21
• /
• 2002

The extract of Ginkgo biloba leaves was measured for inhibitory activity against xanthine oxidase. Aceton extract of G. biloba leaves showed strong inhibitory activity. Inhibitory activities of the fractionated extract were in the order of water > ethyl acetate fractions. Two fractions exhibiting strong inhibitory activities ware further purified via repeated silica gel, Amberlite IRN-78, Polyclar AT, and Sephadex LH-20 column chromatographies. Active components were isolated and identified through $^1$H-NMR and $^{13}$ C-NMR. The compounds were characterized as kaempferol 3-Ο-$\alpha$-(6$^{"′}$-p-coumaroylglucosyl-$\beta$-1,4-rhamnoside), and quercertin 3-Ο-$\alpha$-(6$^{′}$-p-coumaroylglucosyl-$\beta$-1,4-rhamnoside).

• Bulletin of the Korean Chemical Society
• /
• v.30 no.12
• /
• pp.2984-2988
• /
• 2009

A gene (PH0311) encoding a hypothetical protein from the genome sequence data of the hyperthermophilic archaeon Pyrococcus horikoshii OT3 was cloned and over-expressed in Escherichia coli. The purified recombinant protein was found to possess FAD-dependent NADH oxidase activity, although it lacked sequence homology to any other known general NADH oxidase family. The product of the PH0311 gene was thus designated PhNOX (NADH oxidase from Pyrococcus horikoshii), with an estimated molecular weight of 84 kDa by gel filtration and 22 kDa by SDS-PAGE, indicating it to be a homotetramer of 22 kDa subunits. PhNOX catalyzed the oxidation of reduced ${\beta}$-NADH with subsequent formation of $H_2O_2$ in the presence of FAD as a cofactor, but not ${\alpha}$-NADH, ${\alpha}$-NADPH, or ${\beta}$-NADPH. PhNOX showed high affinity for ${\beta}$-NADH with a Km value of 3.70 ${\mu}$M and exhibited optimum activity at pH 8.0 and 95$^{\circ}C$ as it is highly stable against high temperature.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.39 no.4
• /
• pp.481-486
• /
• 2010

The nutraceutical role of omija (Schizandra chinensis Baillon) water extract (OWE) was determined through the analysis of antioxidant activity, nitrite scavening activity, and xanthine oxdiase and $\alpha$-glucosidase inhibitory effects. Antioxidant activity of OWE was measured by using 2,2-diphenyl-1-picryl-hydrazyl (DPPH) free radical scavenging activity and superoxide dismutase-like activity (SODA). DPPH radical scavenging activity and SODA increased in a dose-dependent manner, and was about 49.0% at 2.5 mg/mL and 69.2% at 5 mg/mL, respectively. The xanthine oxidase and $\alpha$-glucosidase inhibitory activities of OWE were about 88.8% and 86.2% at 1 mg/mL, respectively. Nitrite scavenging activity of OWE was about 54.9%, 42.4%, and 34.2% on pH 1.2, 3.0, and 6.0 at 1 mg/mL, respectively. These results suggest that OWE has a strong antioxidant activity, and xanthine oxidase and $\alpha$-glucosidase inhibitory effects.