Screening for microorganisms producing D-malate from maleate

Werf, M.J. van der; Tweel, W.J.J. van den; Hartmans, S.

doi:10.1128/aem.58.9.2854-2860.1992

Cited by 45 publications

(8 citation statements)

References 33 publications

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“…The product of malease activity with maleate iS D-malate (25), and the product of citraconate had the same optical configuration as commercially available D-citramalate (analyzed by optical rotation).…”

Section: Resultsmentioning

confidence: 93%

“…Optical rotation. The optical rotation of the malate and citramalate produced was assayed as described previously (25).…”

Section: Methodsmentioning

confidence: 99%

“…Malease is also present in microorganisms (25), and D-malate production from maleate was studied in more detail in Pseudomonas pseudoalcaligenes NCIMB 9867. This strain produced D-malate with an enantiomeric purity of more than 99.9%.…”

mentioning

confidence: 99%

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Purification and Characterization of Maleate Hydratase from Pseudomonas pseudoalcaligenes

Werf

Tweel

Hartmans

1993

Appl Environ Microbiol

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Maleate hydratase (malease) from Pseudomonas pseudoalcaligenes has been purified. The purified enzyme (98% pure) catalyzes the stereospecific addition ofwater to maleate and citraconate (2-methylmaleate), forming D-(+)-malate and D-(+)-citramalate, respectively. 2,3-Dimethylmaleate was also a substrate for malease. The stability of the enzyme was dependent on the protein concentration and the addition of dicarboxylic acids. The purified enzyme (89 kDa) consisted of two subunits (57 and 24 kDa). No cofactor was required for full activity of this colorless enzyme. Maximum enzyme activity was measured at pH 8 and 45°C. The K,m for maleate was 0.35 mM, and that for citraconate was 0.20 mM. Thiol reagents, such as p-chloromercuribenzoate and iodoacetamide, and sodium dodecyl sulfate completely inhibited malease activity. Malease activity was competitively inhibited by D-malate (Ki = 0.63 mM) and D-citramalate (Ki = 0.083 mM) and by the substrate analog 2,2-dimethylsuccinate (Ki = 0.025 mM). The apparent equilibrium constants for the maleate,

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Section: Resultsmentioning

confidence: 93%

“…Optical rotation. The optical rotation of the malate and citramalate produced was assayed as described previously (25).…”

Section: Methodsmentioning

confidence: 99%

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Purification and Characterization of Maleate Hydratase from Pseudomonas pseudoalcaligenes

Werf

Tweel

Hartmans

1993

Appl Environ Microbiol

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“…In this respect, clear parallels exist to McpP that binds only L-lactate but not D-lactate 36 . Many bacteria are able to synthesize D-malate 68 , 69 and Pseudomonas species were found to metabolize both isomers 70 . However, L-malate is a common carboxylic acid whereas its D-isomer is less frequent 71 .…”

Section: Discussionmentioning

confidence: 99%

The activity of the C4-dicarboxylic acid chemoreceptor of Pseudomonas aeruginosa is controlled by chemoattractants and antagonists

Martín‐Mora

Ortega

Pérez-Maldonado

et al. 2018

Sci Rep

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Chemotaxis toward organic acids has been associated with colonization fitness and virulence and the opportunistic pathogen Pseudomonas aeruginosa exhibits taxis toward several tricarboxylic acid intermediates. In this study, we used high-throughput ligand screening and isothermal titration calorimetry to demonstrate that the ligand binding domain (LBD) of the chemoreceptor PA2652 directly recognizes five C4-dicarboxylic acids with KD values ranging from 23 µM to 1.24 mM. In vivo experimentation showed that three of the identified ligands act as chemoattractants whereas two of them behave as antagonists by inhibiting the downstream chemotaxis signalling cascade. In vitro and in vivo competition assays showed that antagonists compete with chemoattractants for binding to PA2652-LBD, thereby decreasing the affinity for chemoattractants and the subsequent chemotactic response. Two chemosensory pathways encoded in the genome of P. aeruginosa, che and che2, have been associated to chemotaxis but we found that only the che pathway is involved in PA2652-mediated taxis. The receptor PA2652 is predicted to contain a sCACHE LBD and analytical ultracentrifugation analyses showed that PA2652-LBD is dimeric in the presence and the absence of ligands. Our results indicate the feasibility of using antagonists to interfere specifically with chemotaxis, which may be an alternative strategy to fight bacterial pathogens.

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“…39,[102][103][104][105][106] The substrate spectrum of malease isas is the case for fumaraserather narrow, and maleic acid and citraconic acid are accepted best, but small changes in the functional group pattern of these substrates are allowed. [104][105][106] For example chloromaleate and bromomaleate are hydrated to give α-substituted malates (2S,3S)-3-chloromalate and (2S,3S)-3-bromomalate. 107 In industry, Pseudomonas pseudoalcaligenes containing malease is used for the large-scale production of malic acid.…”

Section: View Article Onlinementioning

confidence: 99%