An NAD+-dependent alanine dehydrogenase from a methylotrophic bacterium

Bellion, Edward; Tan, Filemon K.

doi:10.1042/bj2440565

Cited by 35 publications

(16 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Our native, pure preparation of L-alanine dehydrogenase from R. capsulatus ElFl was stable when stored at 4°C in contrast to that partially purified from strain AD2 (30). The molecular properties of the L-alanine dehydrogenase of R. capsulatus ElFl resemble those of the L-alanine dehydrogenase of cyanobacteria (24) and other nonphotosynthetic bacteria (12,22), although, unlike ours, the bacterial L-alanine dehydrogenases are tetramers with subunits of sizes similar to that of R. capsulatus ElFl (3,21,33).…”

Section: Resultsmentioning

confidence: 56%

Purification and properties of L-alanine dehydrogenase of the phototrophic bacterium Rhodobacter capsulatus E1F1

1989

View full text Add to dashboard Cite

In the phototrophic nonsulfur bacterium Rhodobacter capsulatus ElFl, L-alanine dehydrogenase aminating activity functions as an alternative route for ammonia assimilation when glutamine synthetase is inactivated. L-Alanine dehydrogenase deaminating activity participates in the supply of organic carbon to cells growing on L-alanine as the sole carbon source. L-Alanine dehydrogenase is induced in cells growing on pyruvate plus nitrate, pyruvate plus ammonia, or L-alanine under both light-anaerobic and dark-heterotrophic conditions. The enzyme has been purified to electrophoretic and immunological homogeneity by using affinity chromatography with Red-120 agarose. The native enzyme was an oligomeric protein of 246 kilodaltons (kDa) which consisted of six identical subunits of 42 kDa each, had a Stokes' radius of 5.8 nm, an s20 , of 10.1 S, a D20,, of 4.25 x 10-11 m2 s-5, and a frictional quotient of 1.35. The aminating activity was absolutely specific for NADPH, whereas deaminating activity was strictly NAD dependent, with apparent Kms of 0.25 (NADPH), 0.15 (NAD)), 1.25 (L-alanine), 0.13 (pyruvate), and 16 (ammonium) mM. The enzyme was inhibited in vitro by pyruvate or L-alanine and had two sulfhydryl groups per subunit which were essential for both aminating and deaminating activities.

show abstract

Section: Resultsmentioning

confidence: 56%

Purification and properties of L-alanine dehydrogenase of the phototrophic bacterium Rhodobacter capsulatus E1F1

1989

View full text Add to dashboard Cite

show abstract

“…aureus can increase biofilm and explant pH by consuming explant proteins and amino acids as a source of energy. Protein analysis of the spent medium identified some key S. aureus-origin enzymes involved in amino acid metabolism, including 1-pyrroline-5-carboxylate dehydrogenase (involved in proline catabolism [51]), glucosamine-6-phosphate isomerase (converts D-glucosa- mine 6-phosphate to D-fructose 6-phosphate and ammonia [52]), and alanine dehydrogenase 1 (which catalyzes conversion of alanine and NAD ϩ to pyruvate, ammonium, and NADH [53]). The extracellular matrix, which is a binding site of S. aureus (40), contains abundant quantities of collagen, which is rich in proline and alanine, and hyaluronic acid; the latter contains D-glucosamine (54).…”

Section: Discussionmentioning

confidence: 99%

Staphylococcus aureus Induces Hypoxia and Cellular Damage in Porcine Dermal Explants

et al. 2015

View full text Add to dashboard Cite

fWe developed a porcine dermal explant model to determine the extent to which Staphylococcus aureus biofilm communities deplete oxygen, change pH, and produce damage in underlying tissue. Microelectrode measurements demonstrated that dissolved oxygen (DO) in biofilm-free dermal tissue was 4.45 ؎ 1.17 mg/liter, while DO levels for biofilm-infected tissue declined sharply from the surface, with no measurable oxygen detectable in the underlying dermal tissue. Magnetic resonance imaging demonstrated that biofilm-free dermal tissue had a significantly lower relative effective diffusion coefficient (0.26 ؎ 0.09 to 0.30 ؎ 0.12) than biofilm-infected dermal tissue (0.40 ؎ 0.12 to 0.48 ؎ 0.12; P < 0.0001). Thus, the difference in DO level was attributable to biofilm-induced oxygen demand rather than changes in oxygen diffusivity. Microelectrode measures showed that pH within biofilm-infected explants was more alkaline than in biofilm-free explants (8.0 ؎ 0.17 versus 7.5 ؎ 0.15, respectively; P < 0.002). Cellular and nuclear details were lost in the infected explants, consistent with cell death. Quantitative label-free shotgun proteomics demonstrated that both proapoptotic programmed cell death protein 5 and antiapoptotic macrophage migration inhibitory factor accumulated in the infected-explant spent medium, compared with uninfected-explant spent media (1,351-fold and 58-fold, respectively), consistent with the cooccurrence of apoptosis and necrosis in the explants. Biofilm-origin proteins reflected an extracellular matrix-adapted lifestyle of S. aureus. S. aureus biofilms deplete oxygen, increase pH, and induce cell death, all factors that contribute to impede wound healing.

show abstract

“…Although the presence of NAD-dependent alcohol dehydrogenases (ADH; EC 1.1.1.1) has been reported in some methylotrophic bacteria, e. g. in Hyphomicrobium X (Brooke and Attwood 1983) and in organism PAR (Bellion and Wu 1978), this enzyme only appeared to function in ethanol catabolism. All Gram-negative methylotrophic bacteria studied employ a PQQ-containing methanol dehydrogenase (MDH, EC 1.i.99.8; Duine et al 1978) for methanol oxidation.…”

Section: Discussionmentioning

confidence: 99%

Methanol metabolism in thermotolerant methylotrophic Bacillus strains involving a novel catabolic NAD-dependent methanol dehydrogenase as a key enzyme

et al. 1989

View full text Add to dashboard Cite

Abstract. The enzymology of methanol utilization in thermotolerant methytotrophic Bacillus strains was investigated. In all strains an immunological]y related NAD-dependent methanol dehydrogenase was involved in the initial oxidation of methanol. In cells of Bacillus sp. C1 grown under methanol-limiting conditions this enzyme constituted a high percentage of total soluble protein. The methanol dehydrogenase from this organism was purified to homogeneity and characterized. In cell-free extracts the enzyme displayed biphasic kinetics towards methanol, with apparent Km values of 3.8 and 166 mM. Carbon assimilation was by way of the fructose-1,6-bisphosphate aldolase cleavage and transketolase/transaldolase rearrangement variant of the RuMP cycle of formaldehyde fixation. The key enzymes of the RuMP cycle, hexulose-6-phosphate synthase (HPS) and hexulose-6-phosphate isomerase (HPI), were present at very high levels of activity. Failure of whole cells to oxidize formate, and the absence of formaldehyde-and formate dehydrogenases indicated the operation of a non-linear oxidation sequence for formaldehyde via HPS. A comparison of the levels of methanol dehydrogenase and HPS in cells of Bacillus sp. C1 grown on methanol and glucose suggested that the synthesis of these enzymes is not under coordinate control.

show abstract

An NAD+-dependent alanine dehydrogenase from a methylotrophic bacterium

Cited by 35 publications

References 30 publications

Purification and properties of L-alanine dehydrogenase of the phototrophic bacterium Rhodobacter capsulatus E1F1

Purification and properties of L-alanine dehydrogenase of the phototrophic bacterium Rhodobacter capsulatus E1F1

Staphylococcus aureus Induces Hypoxia and Cellular Damage in Porcine Dermal Explants

Methanol metabolism in thermotolerant methylotrophic Bacillus strains involving a novel catabolic NAD-dependent methanol dehydrogenase as a key enzyme

Contact Info

Product

Resources

About