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Identification and Characterization of Some Genes, Enzymes, and Metabolic Intermediates Belonging to the Bile Acid Aerobic Catabolic Pathway from Pseudomonas

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Microbial Steroids

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2704))

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Abstract

The study of the catabolic potential of microbial species isolated from different habitats has allowed the identification and characterization of bacteria able to assimilate bile acids and/or other steroids (e.g., testosterone and 4-androsten-3,17-dione) under aerobic conditions through the 9,10-seco pathway. From soil samples, we have isolated several strains belonging to genus Pseudomonas that grow efficiently in chemically defined media containing some cyclopentane–perhydrophenanthrene derivatives as carbon sources. Genetic and biochemical studies performed with one of these bacteria (P. putida DOC21) allowed the identification of the genes and enzymes belonging to the route involved in bile acids and androgens, the 9,10-seco pathway in this bacterium. In this manuscript, we describe the most relevant methods used in our lab for the identification of the chromosomal location and nucleotide sequence of the catabolic genes (or gene clusters) encoding the enzymes of this pathway, and the tools useful to establish the role of some of the enzymes that participate in this route.

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Acknowledgments

This research was funded by the Ministerio de Economía y Competitividad (Madrid, España, grants BFU2009-11545-C03-01, BIO2012-39695-C02-02, and BIO2015-66960-C3-3R), by a CENIT Project RTC-2014-2249-1 (CDTI, Ministerio de Economía y Competitividad, Madrid, España), and by a grant from the Junta de Castilla y León (Consejería de Educación, Valladolid, España) LE246A11-2. The authors also want to thank the support to their actual research by the Horizon Europe Framework Programme (call: HORIZON-CL4-2021-RESILIENCE-01-11) through the ESTELLA project (“DESign of bio-based Thermoset polymer with rEcycLing capabiLity by dynAmic bonds for bio-composite manufacturing”) (Project no. 101058371), the Ministerio de Ciencia e Innovación (grant TED2021-132593B-I00 belonging to the 2021 convocatory “Proyectos Estratégicos Orientados a la Transición Ecológica y a la Transición Digital” and RTI2018-095584-B-C43 from Proyectos de I+D+i RETOS INVESTIGACION), and the Junta de Castilla y León, grant LE250P20.

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Authors and Affiliations

  1. Departamento de Biología Molecular (Área de Bioquímica y Biología Molecular), Facultad de Veterinaria, Universidad de León, León, Spain

    José M. Luengo & Elias R. Olivera

Authors
  1. José M. Luengo
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  2. Elias R. Olivera
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Correspondence to Elias R. Olivera .

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Editors and Affiliations

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

    Carlos Barreiro

  2. Department of Biotechnology, Curia Spain, León, Spain

    José-Luis Barredo

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Luengo, J.M., Olivera, E.R. (2023). Identification and Characterization of Some Genes, Enzymes, and Metabolic Intermediates Belonging to the Bile Acid Aerobic Catabolic Pathway from Pseudomonas. In: Barreiro, C., Barredo, JL. (eds) Microbial Steroids. Methods in Molecular Biology, vol 2704. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3385-4_4

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  • DOI: https://doi.org/10.1007/978-1-0716-3385-4_4

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3384-7

  • Online ISBN: 978-1-0716-3385-4

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