1887

Abstract

can utilize proteogenic amino acids as the sole source of carbon and nitrogen. In particular, utilization of -Asp and -Asn is insensitive to carbon catabolite repression as strong growth remains in the mutants devoid of the essential CbrAB activators of most catabolic genes. Transcriptome analysis was conducted to identify genes for the catabolism, uptake and regulation of these two amino acids. Gene inactivation and growth phenotype analysis established two asparaginases AsnA and AsnB for the degradation of -Asn to -Asp, whereas only AnsB is required for the deamidation of -Asn to -Asp. While -Asp is a dead-end product, conversion of -Asp to fumarate is catalysed by an aspartase AspA as further evidenced by enzyme kinetics. The results of measuring promoter- expression and mobility shift assays demonstrated that and encode two transcriptional regulators in response to -Asn and -Asp, respectively, for the induction of the operon and the gene. Exogenous -Glu also caused induction of the gene, most likely due to its conversion to -Asp by the aspartate transaminase AspC. Expression of several transporters were found inducible by -Asn and/or -Asp, including AatJQMP for acid amino acids, DctA and DctPQM for C4-dicarboxylates, and PA5530 for C5-dicarboxylates. In summary, a complete pathway and regulation for -Asn and -Asp catabolism was established in this study. Cross induction of three transport systems for dicarboxylic acids may provide a physiological explanation for the insensitivity of -Asn and -Asp utilization to carbon catabolite repression.

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2018-02-01
2020-10-24
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References

  1. Nishijyo T, Haas D, Itoh Y. The CbrA-CbrB two-component regulatory system controls the utilization of multiple carbon and nitrogen sources in Pseudomonas aeruginosa. Mol Microbiol 2001;40:917–931 [CrossRef][PubMed]
    [Google Scholar]
  2. Li W, Lu CD. Regulation of carbon and nitrogen utilization by CbrAB and NtrBC two-component systems in Pseudomonas aeruginosa. J Bacteriol 2007;189:5413–5420 [CrossRef][PubMed]
    [Google Scholar]
  3. Singh B, Röhm KH. Characterization of a Pseudomonas putida ABC transporter (AatJMQP) required for acidic amino acid uptake: biochemical properties and regulation by the Aau two-component system. Microbiology 2008;154:797–809 [CrossRef][PubMed]
    [Google Scholar]
  4. Sonawane AM, Singh B, Röhm KH. The AauR-AauS two-component system regulates uptake and metabolism of acidic amino acids in Pseudomonas putida. Appl Environ Microbiol 2006;72:6569–6577 [CrossRef][PubMed]
    [Google Scholar]
  5. Sonawane AM, Röhm KH. A functional gltB gene is essential for utilization of acidic amino acids and expression of periplasmic glutaminase/asparaginase (PGA) by Pseudomonas putida KT2440. Mol Genet Genomics 2004;271:33–39 [CrossRef][PubMed]
    [Google Scholar]
  6. Sonawane A, Klöppner U, Hövel S, Völker U, Röhm KH. Identification of Pseudomonas proteins coordinately induced by acidic amino acids and their amides: a two-dimensional electrophoresis study. Microbiology 2003;149:2909–2918 [CrossRef][PubMed]
    [Google Scholar]
  7. Sonawane A, Klöppner U, Derst C, Röhm KH. Utilization of acidic amino acids and their amides by pseudomonads: role of periplasmic glutaminase-asparaginase. Arch Microbiol 2003;179:151–159 [CrossRef][PubMed]
    [Google Scholar]
  8. He W, Li G, Yang CK, Lu CD. Functional characterization of the dguRABC locus for D-Glu and d-Gln utilization in Pseudomonas aeruginosa PAO1. Microbiology 2014;160:2331–2340 [CrossRef][PubMed]
    [Google Scholar]
  9. He W, Li C, Lu CD. Regulation and characterization of the dadRAX locus for d-amino acid catabolism in Pseudomonas aeruginosa PAO1. J Bacteriol 2011;193:2107–2115 [CrossRef][PubMed]
    [Google Scholar]
  10. Li C, Yao X, Lu CD. Regulation of the dauBAR operon and characterization of D-amino acid dehydrogenase DauA in arginine and lysine catabolism of Pseudomonas aeruginosa PAO1. Microbiology 2010;156:60–71 [CrossRef][PubMed]
    [Google Scholar]
  11. Li C, Lu CD. Arginine racemization by coupled catabolic and anabolic dehydrogenases. Proc Natl Acad Sci USA 2009;106:906–911 [CrossRef][PubMed]
    [Google Scholar]
  12. Li G, Lu CD. Molecular characterization of LhpR in control of hydroxyproline catabolism and transport in Pseudomonas aeruginosa PAO1. Microbiology 2016;162:1232–1242 [CrossRef][PubMed]
    [Google Scholar]
  13. Li G, Lu CD. The cryptic dsdA gene encodes a functional D-serine dehydratase in Pseudomonas aeruginosa PAO1. Curr Microbiol 2016;72:788–794 [CrossRef][PubMed]
    [Google Scholar]
  14. Haas D, Holloway BW, Schamböck A, Leisinger T. The genetic organization of arginine biosynthesis in Pseudomonas aeruginosa. Mol Gen Genet 1977;154:7–22 [CrossRef][PubMed]
    [Google Scholar]
  15. Weiner B, Poelarends GJ, Janssen DB, Feringa BL. Biocatalytic enantioselective synthesis of N-substituted aspartic acids by aspartate ammonia lyase. Chemistry 2008;14:10094–10100 [CrossRef][PubMed]
    [Google Scholar]
  16. Valentini M, Lapouge K. Catabolite repression in Pseudomonas aeruginosa PAO1 regulates the uptake of C4 -dicarboxylates depending on succinate concentration. Environ Microbiol 2013;15:1707–1716 [CrossRef][PubMed]
    [Google Scholar]
  17. Valentini M, Storelli N, Lapouge K. Identification of C(4)-dicarboxylate transport systems in Pseudomonas aeruginosa PAO1. J Bacteriol 2011;193:4307–4316 [CrossRef][PubMed]
    [Google Scholar]
  18. Takagi JS, Tokushige M, Shimura Y. Cloning and nucleotide sequence of the aspartase gene of Pseudomonas fluorescens. J Biochem 1986;100:697–705 [CrossRef][PubMed]
    [Google Scholar]
  19. Lundgren BR, Villegas-Peñaranda LR, Harris JR, Mottern AM, Dunn DM et al. Genetic analysis of the assimilation of C5-dicarboxylic acids in Pseudomonas aeruginosa PAO1. J Bacteriol 2014;196:2543–2551 [CrossRef][PubMed]
    [Google Scholar]
  20. Gelfand DH, Steinberg RA. Escherichia coli mutants deficient in the aspartate and aromatic amino acid aminotransferases. J Bacteriol 1977;130:429–440[PubMed]
    [Google Scholar]
  21. Gu W, Song J, Bonner CA, Xie G, Jensen RA. PhhC is an essential aminotransferase for aromatic amino acid catabolism in Pseudomonas aeruginosa. Microbiology 1998;144:3127–3134 [CrossRef][PubMed]
    [Google Scholar]
  22. Castanie-Cornet MP, Penfound TA, Smith D, Elliott JF, Foster JW. Control of acid resistance in Escherichia coli. J Bacteriol 1999;181:3525–3535[PubMed]
    [Google Scholar]
  23. Hoang TT, Karkhoff-Schweizer RR, Kutchma AJ, Schweizer HP. A broad-host-range Flp-FRT recombination system for site-specific excision of chromosomally-located DNA sequences: application for isolation of unmarked Pseudomonas aeruginosa mutants. Gene 1998;212:77–86 [CrossRef][PubMed]
    [Google Scholar]
  24. Farinha MA, Kropinski AM. Construction of broad-host-range plasmid vectors for easy visible selection and analysis of promoters. J Bacteriol 1990;172:3496–3499 [CrossRef][PubMed]
    [Google Scholar]
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