• Microbiology and Biotechnology Letters
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• v.47 no.1
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• pp.139-147
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• 2019

Glutamate decarboxylase catalyzes the conversion of glutamate to gamma-aminobutyric acid (GABA), contributing to pH homeostasis through proton consumption. The reaction is the first step toward the GABA shunt. To date, the enzymes involved in the glutamate metabolism of Photobacterium damselae subsp. piscicida have not been elucidated. In this study, an open reading frame of P. damselae subsp. piscicida, showing homology to the glutamate decarboxylase or putative pyridoxal-dependent aspartate 1-decarboxylase genes, was isolated and cloned into an expression vector to produce the recombinant enzyme. Preliminary gas chromatography-mass spectrometry characterization of the purified recombinant enzyme revealed that it catalyzed not only the decarboxylation of glutamate but also the transamination of GABA. This enzyme of P. damselae subsp. piscicida could be bifunctional, combining decarboxylase and transaminase activities in a single polypeptide chain.

• The Plant Pathology Journal
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• v.39 no.3
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• pp.235-244
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• 2023

Acidovorax citrulli (Ac) is a phytopathogenic bacterium that causes bacterial fruit blotch (BFB) in cucurbit crops, including watermelon. However, there are no effective methods to control this disease. YggS family pyridoxal phosphate-dependent enzyme acts as a coenzyme in all transamination reactions, but its function in Ac is poorly understood. Therefore, this study uses proteomic and phenotypic analyses to characterize the functions. The Ac strain lacking the YggS family pyridoxal phosphate-dependent enzyme, AcΔyppAc(EV), virulence was wholly eradicated in geminated seed inoculation and leaf infiltration. AcΔyppAc(EV) propagation was inhibited when exposed to L-homoserine but not pyridoxine. Wild-type and mutant growth were comparable in the liquid media but not in the solid media in the minimal condition. The comparative proteomic analysis revealed that YppAc is primarily involved in cell motility and wall/membrane/envelop biogenesis. In addition, AcΔyppAc(EV) reduced biofilm formation and twitching halo production, indicating that YppAc is involved in various cellular mechanisms and possesses pleiotropic effects. Therefore, this identified protein is a potential target for developing an efficient anti-virulence reagent to control BFB.

• Parasites, Hosts and Diseases
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• v.21 no.1
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• pp.49-57
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• 1983

The distribution and Properties of branched chain amino acid aminotransferase (EC 2.6. 1.42) was investigated in adult Fasciola hepatica. Fascicla hepatica was fractionated by differential centrifugation into nuclear, mitochondrial and cytosolic fractions. The activity of branched chain amino acid aminotransferase was measured by the method of Ichihara and Koyama (1966) . Isozyme patterns of this enzyule was also examined by DEAE-cellulose column chromatography. The results obtained were as follows; 1. The activity in homogenate was found to be 12.69 units/g wet tissue. The activity of this enzyme was relatively high compared with those in rat tissues. 2. The distribution of branched chain amino acid aminotransferase in the subcellular organelles showed that 87.8% of the activity was in cytosolic, 10.9% in mitochondrial and 1.3% was in nuclear fraction. 3. Cytosolic fraction of Fasciola hepatica contained Enzyme I, but not Enzyme II and III, of branched chain amino acid aminotransferase. Ensyme I was eluted by 50mM phosphate buffier from DEAE-cellulose column and catalyzed the transamination of all three branched chain amino acids. 4. The Enzyme I was purified about 22-folds increase in specific activity after chromatography on DEAE-cellulose. 5. The best substrate among three amino acids (leucine, isoleucine and valise) was L-isoleucine. 6. The optimal temperature of Enzyme I was $45^{\circ}C$ and the optimal pH was 8.2. 7. The Km value for leucine of Enzyme I was 4.17 mM. 8. The Km values for a-ketoglutarate and pyridoxal phosphate of Enzyme I were 0.41mM and $4.76{\times}10^{-3}{\;}mM$, respectively.

• Journal of Microbiology and Biotechnology
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• v.9 no.6
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• pp.695-703
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• 1999

We have studied the stereospecificities of various pyridoxal 5'-phosphate (PLP) dependent enzymes for the hydrogen transfer between the C-4' of a bound coenzyme and the C-2 of a substrate in the transamination catalyzed by the enzymes. Stereospecificities reflect the structures of enzyme active-sites, in particular the geometrical relationship between the coenzyme-substrate Schiff base and the active site base participating in an $\alpha$-hydrogen abstraction. The PLP enzymes studied so far catalyze only a si-face specific (pro-S) hydrogen transfer. This stereochemical finding suggests that the PLP enzymes have the same topological active-site structures, and that the PLP enzymes have evolved divergently from a common ancestral protein. However, we found that o-amino acid aminotransferase, branched chain L-amino acid aminotransferase, and 4-amino-4-deoxychorismate lyase, which have significant sequence homology with one another, catalyze a re-face specific (pro-R) hydrogen transfer. We also showed that PLP-dependent amino acid racemases, which have no sequence homology with any aminotransferases, catalyze a non-stereospecific hydrogen transfer: the hydrogen transfer occurs on both faces of the planar intermediate. Crystallographical studies have shown that the catalytic base is situated on the re-face of the C-4' of the bound coenzyme in o-amino acid aminotransferase and branched chain L-amino acid aminotransferase, whereas the catalytic base is situated on the si-face in other aminotransferases (such as L-aspartate aminotransferase) catalyzing the si-face hydrogen transfer. Thus, we have clarified the stereospecificities of PLP enzymes in relation with the primary structures and three-dimensional structures of the enzymes. The characteristic stereospecificities of these enzymes for the hydrogen transfer suggest the convergent evolution of PLP enzymes.

• Journal of the Korean Society of Food Science and Nutrition
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• v.13 no.4
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• pp.437-443
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• 1984

Muscular exercise induced by electrical stimulation of femoral nerves in perfused rat hindquarters(5 contractions per sec) in the presence of insulin and glucose effected a rapid increase(c. a. two-fold) in the level of citric acid cycle intermediates. The highest values were found within one minute of stimulation. The tissue concent ratios of lactate, pyruvate and alanine increased rapidly on initiation of exercese. Release of lactate also increased rapidly, whereas that of pyruvate was only moderately elevated. In the course of three minute exercese, the sum of alanine, glutamate and aspartate was only transiently elevated. A fall in creatine-p and ATP in the stimulated muscle was accompanied by increases in tissue level of AMP and release of ammonia into perfusing medium. However, the changes in glutamine were small. It is concluded that the pool of citric acid cycle intermediates is expanded during muscular work due (a) to an elevated level of pyruvate, leading to shifts in the levels of alanine and cycle intermediates vie trans-amination reactions and (b) to stimulation of the purine nucleotide cycle due to elevated AMP, resulting in generation of cycle intermediates and ammonia at the expense of aspartate.

• Fisheries and Aquatic Sciences
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• v.22 no.7
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• pp.15.1-15.8
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• 2019

Corexit 9500 is a dispersant commercially available in Nigeria that is used to change the inherent chemical and physical properties of oil, thereby changing the oil's transport and fate with potential effects on the environment. The aim of this study was to assess the biochemical (enzymes and electrolyte) toxicity of Corexit 9500 dispersant on the gills, liver and kidney of juveniles of Clarias gariepinus after exposure for 21 days. One hundred sixty fish were used without gender consideration. Range-finding tests were conducted over a 96-h period after acclimatisation of the test organisms in the laboratory. The test organisms (10/treatment) were exposed to Corexit 9500 in the following concentrations-0.00, 0.0125, 0.025 and 0.05 ml/l in triplicate. Twenty-one days later, fish was dissected. 0.5 g from each of the following organs-gills, liver and kidney tissues-was removed, homogenised and tested for enzymes [superoxide dismutase (SOD), catalase (CAT), alanine aminotransferase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP)], urea, creatinine and electrolytes (sodium ($Na^+$), potassium ($K^+$), chloride ($Cl^-$), bicarbonate ($HCO_3{^-}$)) following standard methods. In the gills, SOD and ALT to AST ratio were significantly lower than in control while the creatinine was significantly higher in the toxicant. In the kidney, creatinine was significantly higher in fish exposed to the toxicant. In the liver, ALP increased in the toxicant while urea was decreased. The mean electrolyte concentrations ($Na^+$, $K^+$, $Cl^-$ and $HCO_3{^-}$) increased significantly in the concentration of the toxicant (P < 0.05). The alterations observed in the activities of these electrolytes and enzymes indicated that Corexit 9500 interfered with transamination and metabolic functions of the fish.

• The Korean Journal of Zoology
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• v.17 no.3
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• pp.131-138
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• 1974

Sera from male Spague-Dawley rats, exposed to $30\\pm 0.5^\\circ C$ for 240 hours or $33\\pm 0.5^\\circ C$ for 64 hours, were assayed for the activities of serum glutamic oxaloacetic transaminase(SGOT) and serum glutamic pyruvic transaminase(SGPT) at various time during the heat exposure. 1. When compared to control animals maintained at $23\\pm 1^\\circ C$, the animals exposed to $30\\pm 0.5^\\circ C$ ro $33\\pm 0.5^\\circ C$ showed a significant increase in SGOT and SGPT activities, 2. The SGOT activity incressed at 16 and 72 hours after the exposure to $30^\\circ C$, and at 30 and 64 hours after the exposure to $33^\\circ C$. After 72 hours, the activity returned to the initial value in case of $30^\\circ C$ exposure. 3. The SGPT activity increased significantly as early as 4 hours after the exposure to $30^\\circ C$ or $33^\\circ C$. It was also high at 16 hours after the exposure. The activity was also high at 72 hours and at 64 hours after the exposure to $30^\\circ C$ and $33^\\circ C$ respectively. After 144 hours, SGPT level increased slightly in the case of $30^\\circ C$ exposure. 4. The activities of SGOT and SGPT were significantly higher in rats exposed to $33^\\circ C$ at 16, 30, and 64 hours than those exposed to $30^\\circ C$. 5. It may be inferred from above data that the prolonged heat exposed rat has the abnormal metabolism of transamination.