Biochemical characterization of two glutamate dehydrogenases with different cofactor specificities from a hyperthermophilic archaeon Pyrobaculum calidifontis

Wakamatsu, Taisuke; Higashi, Chisato; Ohmori, Taketo; Doi, Kunio; Ohshima, Toshihisa

doi:10.1007/s00792-013-0527-7

Cited by 7 publications

(3 citation statements)

References 41 publications

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“…Discussion. Data on inhibitory of NADP +dependent glutamate dehydrogenase A. cal coaceticus ІМV B-7241 activity with potassium and sodium cations in relatively high concentrations (over 50 mM, see Table 1) are consistent with the literature data [13,14]. Thus, in the presence of 1 M of potassium chloride, a 30 % decrease in the activity of the purified enzyme was observed in hyperthermophilic archaea Thermococcus waiotapuensis [13], the activity of purified NADP + -dependent glutamate dehydrogenase of Pyrobaculum calidifontis was inhibited by 50 % in the presence of 100-200 mM of potassium chloride and 100-300 mM of sodium chloride [14].…”

Section: Abstract: Acinetobacter Calcoaceticus Imv B-7241 Surfactants Potassium and Sodium Cations Biological Activity Biosynthesissupporting

confidence: 88%

“…It is known from the literature that monovalent cations can be both inhibitors [13,14] and activators of NADP + -glutamate dehydrogenase [7,8]. In paper [15] it is noted that the stability of Escherichia coli glutamate dehydrogenase increased in the presence of lithium cations at concentrations from 1 to 10 mM.…”

Section: Abstract: Acinetobacter Calcoaceticus Imv B-7241 Surfactants Potassium and Sodium Cations Biological Activity Biosynthesismentioning

confidence: 99%

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Synthesis and Biological Activity of Acinetobacter calcoaceticus IMV B-7241 Surfactants Depending on Monovalent Cations Content in Cultivation Medium

Пирог¹,

Lutsai²,

Shevchuk³

et al. 2021

Mikrobiol. Z.

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Microbial surfactants (biosurfactants) are multifunctional preparations due to a combination of physicochemical (reduction of surface and interfacial tension, emulsifying activity) and biological (antimicrobial and antiadhesive activity, the ability to destroy biofilms) properties. However, the disadvantage of biosurfactants synthesized as a complex of compounds is the possibility of changing the biological activity depending on the conditions of producer cultivation. Aim. To study the effect of potassium and sodium cations on the NADP+-dependent glutamate dehydrogenase activity of cell-free extract of Acinetobacter calcoaceticus ІМV B-7241 with subsequent appropriate modification of the nutrient medium composition and determination of antimicrobial and anti-adhesive activity of surfactant synthesized. Methods. A. calcoaceticus ІМV B-7241 strain was grown in media containing 2% of sunflower oil waste as a carbon source, as well as various concentrations of potassium and sodium chloride (basal – 1.0 g/l NaCl, medium #1 that did not contains NaCl, medium #2 in which the concentration of NaCl was 2.0 g/l, medium #3 in which the concentration of NaCl and KCl was 1.0 g/l each). The surfactants were extracted from the supernatant liquid culture with a modified Folch mixture. Antiadhesive activity and the degree of biofilms degradation were determined by spectrophotometric method, antimicrobial activity − by the indicator of the minimum inhibitory concentration (MIC). Activity of enzymes of surface-active aminolipids biosynthesis (NADP+-dependent glutamate dehydrogenase) and glycolipids (phosphoenolpyruvate (PEP) carboxylase, PEP-synthetase, PEP-carboxykinase, trehalose phosphate synthase) were analyzed in cell-free extracts obtained after the destruction of cells by ultrasound. Results. It was found that potassium and sodium cations in concentrations of 50 and 100 mM are inhibitors of NADP+-dependent glutamate dehydrogenase, and in lower concentrations (5–20 mM) – activators of this enzyme, as well as PEP-carboxykinase and PEP-synthetase. The increase in the biosurfactant concentration to 6.1−7.7 g/l during cultivation of A. calcoaceticus ІМV B-7241 in medium #1 and #3 was due to the predominant synthesis of glycolipids under such conditions, which was evidenced by the increase in 1.8−6.5 times in the activity of PEP-carboxylase, PEP-carboxykinase, PEP-synthetase and trehalose phosphate synthetase compared to the indicators on the basal medium. The concentration of surfactants synthesized in the basal medium was 3.6 g/l, but such surfactants were characterized by the highest antimicrobial and anti-adhesive activity. Their MIC against the test-cultures of studied bacteria (Pseudomonas sp. MI-2, Bacillus subtilis BT-2, Escherichia coli IEM-1, Staphylococcus aureus BMS-1, Enterobacter cloaceae C-8) and fungi (Candida albicans D-6, Rhizopus nigricans P1, Aspergillus niger P-3, Fusarium culmorum T-7) were 0.88−56 μg/ml and were by 2−3 orders of magnitude lower compared to established for surfactants synthesized in modified media #1–3. In the case of treatment of abiotic materials with surfactant solutions obtained on the basal medium, the adhesion of bacteria and fungi was on average 10–20% lower than after surface treatment by the surfactant synthesized in modified media. In the presence of 148−296 μg/ml of surfactants obtained in the basal medium, destruction of S. aureus BMS-1 and B. subtilis BT-2 biofilms was 45−66%, and C. albicans D-6 yeast – 39−44%. Under the action of similar concentrations of surfactants synthesized in modified media, the destruction of bacterial and yeast biofilms was lower: 6-52 and 20–46%, respectively. Conclusions. The obtained results are consistent with the data of our previous studies on the possibility of regulating the antimicrobial and antiadhesive activity of surfactants in the process of producer cultivation by changing the content of cations in the medium, which are inhibitors/activators of enzymes responsible for the synthesis of components of the surfactants complex, which have certain biological properties.

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Section: Abstract: Acinetobacter Calcoaceticus Imv B-7241 Surfactants Potassium and Sodium Cations Biological Activity Biosynthesissupporting

confidence: 88%

Section: Abstract: Acinetobacter Calcoaceticus Imv B-7241 Surfactants Potassium and Sodium Cations Biological Activity Biosynthesismentioning

confidence: 99%

Synthesis and Biological Activity of Acinetobacter calcoaceticus IMV B-7241 Surfactants Depending on Monovalent Cations Content in Cultivation Medium

Пирог¹,

Lutsai²,

Shevchuk³

et al. 2021

Mikrobiol. Z.

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“…The K m of glutamate is comparable with the reported K m values of glutamate for GDH obtained from thermophilic microbial species, which range from 0.025 to over 25,000 μM (56-59). However, our calculated K m for NAD + is slightly higher than the range reported for thermophilic species (18-350 μM) (56)(57)(58)(59).…”

Section: Applicability Of the 24-dnph α-Kg Assay To Non-phd Enzymescontrasting

confidence: 83%

Development of a colorimetric α-ketoglutarate detection assay for prolyl hydroxylase domain (PHD) proteins

Wong

Ringel

Yuan

et al. 2021

Journal of Biological Chemistry

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Since the discovery of the prolyl hydroxylases domain (PHD) proteins and their canonical hypoxia-inducible factor (HIF) substrate two decades ago, a number of in vitro hydroxylation (IVH) assays for PHD activity have been developed to measure the PHD–HIF interaction. However, most of these assays either require complex proteomics mass spectrometry methods that rely on the specific PHD–HIF interaction or require the handling of radioactive material, as seen in the most commonly used assay measuring [ 14 C]O 2 release from labeled [ 14 C]α-ketoglutarate. Here, we report an alternative rapid, cost-effective assay in which the consumption of α-ketoglutarate is monitored by its derivatization with 2,4-dinitrophenylhydrazine (2,4-DNPH) followed by treatment with concentrated base. We extensively optimized this 2,4-DNPH α-ketoglutarate assay to maximize the signal-to-noise ratio and demonstrated that it is robust enough to obtain kinetic parameters of the well-characterized PHD2 isoform comparable with those in published literature. We further showed that it is also sensitive enough to detect and measure the IC 50 values of pan-PHD inhibitors and several PHD2 inhibitors in clinical trials for chronic kidney disease (CKD)-induced anemia. Given the efficiency of this assay coupled with its multiwell format, the 2,4-DNPH α-KG assay may be adaptable to explore non-HIF substrates of PHDs and potentially to high-throughput assays.

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Two different alanine dehydrogenases from Geobacillus kaustophilus: Their biochemical characteristics and differential expression in vegetative cells and spores

Maeno

Ohmori

Nukada

et al. 2023

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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Biochemical characterization of two glutamate dehydrogenases with different cofactor specificities from a hyperthermophilic archaeon Pyrobaculum calidifontis

Cited by 7 publications

References 41 publications

Synthesis and Biological Activity of Acinetobacter calcoaceticus IMV B-7241 Surfactants Depending on Monovalent Cations Content in Cultivation Medium

Synthesis and Biological Activity of Acinetobacter calcoaceticus IMV B-7241 Surfactants Depending on Monovalent Cations Content in Cultivation Medium

Development of a colorimetric α-ketoglutarate detection assay for prolyl hydroxylase domain (PHD) proteins

Two different alanine dehydrogenases from Geobacillus kaustophilus: Their biochemical characteristics and differential expression in vegetative cells and spores

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