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Production of 3-hydroxy-n-phenylalkanoic acids by a genetically engineered strain of Pseudomonas putida

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Abstract

Overexpression of the gene encoding the poly-3-hydroxy-n-phenylalkanoate (PHPhA) depolymerase (phaZ) in Pseudomonas putida U avoids the accumulation of these polymers as storage granules. In this recombinant strain, the 3-OH-acyl-CoA derivatives released from the different aliphatic or aromatic poly-3-hydroxyalkanoates (PHAs) are catabolized through the β-oxidation pathway and transformed into general metabolites (acetyl-CoA, succinyl-CoA, phenylacetyl-CoA) or into non-metabolizable end-products (cinnamoyl-CoA). Taking into account the biochemical, pharmaceutical and industrial interest of some PHA catabolites (i.e., 3-OH-PhAs), we designed a genetically engineered strain of P. putida U (P. putida U ΔfadBA-phaZ) that efficiently bioconverts (more than 80%) different n-phenylalkanoic acids into their 3-hydroxyderivatives and excretes these compounds into the culture broth.

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Acknowledgements

This investigation was supported by the Comisión Interministerial de Ciencia y Tecnología (CICYT), Madrid, Spain (grant BIO2003-05309-C04-01). A.S. and E.A. are recipients of fellowships from the Universidad de León and CICYT, respectively.

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Authors and Affiliations

  1. Departamento de Bioquímica y Biología Molecular, Facultad de Veterinaria, Universidad de León, 24007, Leon, Spain

    Ángel Sandoval, Elsa Arias-Barrau, Elías R. Olivera & José M. Luengo

  2. Departamento de Química Orgánica, Facultad de Químicas, Universidad de Salamanca, Salamanca, Spain

    Francisco Bermejo

  3. Instituto Biomar, Onzonilla, Leon, Spain

    Librada Cañedo

  4. Departamento de Patología Animal (Sanidad Animal), Facultad de Veterinaria, Universidad de León, Leon, Spain

    Germán Naharro

Authors
  1. Ángel Sandoval
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  2. Elsa Arias-Barrau
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  3. Francisco Bermejo
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  4. Librada Cañedo
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  5. Germán Naharro
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  6. Elías R. Olivera
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  7. José M. Luengo
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Corresponding author

Correspondence to José M. Luengo.

Appendix

Appendix

NMR data for 3-hydroxy-5-phenylpentanoic acid

1H NMR (CDCl3): δ=1.74 (m; 2H, H-4) 2.45 (m; 2H, H-2), 2.74 (m; 2H, H-5), 4.05 (s, broad; 1H, H-3), 7.17–7.27 (m; 5H, aromatic H). 13C NMR (CDCl3): δ=32.09 (C-5), 37.56 (C-4), 41.27 (C-2), 67.38(C-3), 125.60 (C-4′), 128.31(C-2′, C-6′), 128.21 (C-3′, C-5′), 141.55 (C-1′), 175.97 (C-1).

NMR data for 3-hydroxy-6-phenylhexanoic acid

1H NMR (CDCl3): δ=1.55 (m; 2H, H-4), 1.68–1.81 (m; 2H, H-5), 2.54 (m; 2H, H-2), 2.66 (m; 2H, H-6), 4.08 (s, broad; 1H, H-3), 7.30–7.20 (m; 5H, aromatic H). 13C NMR (CDCl3): δ=27.50 (C-5), 35.86 (C-6), 36.12 (C-4), 41.41 (C-2), 68.20 (C-3), 126.10 (C-4′), 128.67 (C-2′, C-6′), 128.62 (C-3′, C-5′), 142.32 (C-1′), 178.39 (C-1).

NMR data for 3-hydroxy-7-phenylheptanoic acid

1H NMR (CDCl3): δ=1.39–1.49 (m, 2H, H-5), 1.53 (m; 2H, H-4), 1.65 (m, 2H, H-6), 2.47 (m; 2H, H-2), 2.62 (m; 2H, H-7), 4.05 (s, broad; 1H, H-3), 7.17–7.27 (m; 5H, aromatic H). 13C NMR (CDCl3): δ=25.10 (C-5), 31.30 (C-6), 35.79 (C-7), 36.28 (C-4), 41.18 (C-2), 68.04(C-3), 125.66 (C-4′), 128.36 (C-2′, C-6′), 128.26 (C-3′, C-5′), 142.45 (C-1′), 176.04 (C-1).

NMR data for 3-hydroxy-8-phenyloctanoic acid

1H NMR (CDCl3): δ=1.36 (m; 2H, H-6), 1.37–1.47 (m; 2H, H-5), 1.47–1.53 (m; 2H, H-4), 1.63 (m; 2H, H-7), 2.51 (m; 2H, H-2), 2.63 (m; 2H, H-8), 4.03 (s, broad; 1H, H-3), 7.18–7.28 (m; 5H, aromatic H). 13C NMR (CDCl3): δ=25.29 (C-5), 29.07 (C-6), 31.35 (C-7), 35.83 (C-8), 36.30 (C-4), 41.09 (C-2), 68.02 (C-3), 125.63 (C-4′), 128.39 (C-2′, C-6′), 128.25 (C-3′, C-5′), 142.62 (C-1′), 178.06 (C-1).

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Sandoval, Á., Arias-Barrau, E., Bermejo, F. et al. Production of 3-hydroxy-n-phenylalkanoic acids by a genetically engineered strain of Pseudomonas putida. Appl Microbiol Biotechnol 67, 97–105 (2005). https://doi.org/10.1007/s00253-004-1752-x

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