title: Pseudomonas putida as a platform for the synthesis of aromatic compounds.
abstract: Aromatic compounds such as l-phenylalanine, 2-phenylethanol and trans-cinnamate are aromatic compounds of industrial interest. Current trends support replacement of chemical synthesis of these compounds by 'green' alternatives produced in microbial cell factories. The solvent-tolerant Pseudomonas putida DOT-T1E strain was genetically modified to produce up to 1 g l-1 of l-phenylalanine. In order to engineer this strain, we carried out the following stepwise process: (1) we selected random mutants that are resistant to toxic phenylalanine analogues; (2) we then deleted up to five genes belonging to phenylalanine metabolism pathways, which greatly diminished the internal metabolism of phenylalanine; and (3) in these mutants, we overexpressed the pheAfbr gene, which encodes a recombinant variant of PheA that is insensitive to feedback inhibition by phenylalanine. Furthermore, by introducing new genes, we were able to further extend the diversity of compounds produced. Introduction of histidinol phosphate transferase (PP_0967), phenylpyruvate decarboxylase (kdc) and an alcohol dehydrogenase (adh) enabled the strain to produce up to 180 mg l-1 2-phenylethanol. When phenylalanine ammonia lyase (pal) was introduced, the resulting strain produced up to 200 mg l-1 of trans-cinnamate. These results demonstrate that P. putida can serve as a promising microbial cell factory for the production of l-phenylalanine and related compounds.
authors.affiliation: Biotechnology - CPA Department, Abengoa Research, C/Energía Solar 1, Palmas Altas, Seville, Spain.
authors.forename: Carlos
authors.initials: C
authors.lastname: Molina-Santiago
authors.affiliation: Biotechnology - CPA Department, Abengoa Research, C/Energía Solar 1, Palmas Altas, Seville, Spain.
authors.forename: Baldo F
authors.initials: BF
authors.lastname: Cordero
authors.affiliation: Biotechnology - CPA Department, Abengoa Research, C/Energía Solar 1, Palmas Altas, Seville, Spain.
authors.forename: Abdelali
authors.initials: A
authors.lastname: Daddaoua
authors.affiliation: Biotechnology - CPA Department, Abengoa Research, C/Energía Solar 1, Palmas Altas, Seville, Spain.
authors.forename: Zulema
authors.initials: Z
authors.lastname: Udaondo
authors.affiliation: Biotechnology - Process Development Department, Abengoa Research, Babilafuente, Salamanca, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Manzano
authors.affiliation: Biotechnology - CPA Department, Abengoa Research, C/Energía Solar 1, Palmas Altas, Seville, Spain.
authors.forename: Miguel
authors.initials: M
authors.lastname: Valdivia
authors.affiliation: Biotechnology - CPA Department, Abengoa Research, C/Energía Solar 1, Palmas Altas, Seville, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Segura
authors.affiliation: Biotechnology - Process Development Department, Abengoa Research, Babilafuente, Salamanca, Spain.|Biotechnology - CPA Department, Abengoa Research, C/Energía Solar 1, Palmas Altas, Seville, Spain.
authors.forename: Juan-Luis
authors.initials: JL
authors.lastname: Ramos
authors.affiliation: Biotechnology - CPA Department, Abengoa Research, C/Energía Solar 1, Palmas Altas, Seville, Spain.
authors.forename: Estrella
authors.initials: E
authors.lastname: Duque
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002934:Cinnamates / Q000378:metabolism*; D005421:Flavoring Agents / Q000378:metabolism*; D007218:Industrial Microbiology; D010649:Phenylalanine / Q000096:biosynthesis*; D010626:Phenylethyl Alcohol / Q000378:metabolism*; D016958:Pseudomonas putida / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D002934:Cinnamates; D005421:Flavoring Agents; D010649:Phenylalanine; D010626:Phenylethyl Alcohol