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Author (up) Donoso, R.A.; Ruiz, D.; Garate-Castro, C.; Villegas, P.; Gonzalez-Pastor, J.E.; de Lorenzo, V.; Gonzalez, B.; Perez-Pantoja, D.
Title Identification of a self-sufficient cytochrome P450 monooxygenase from Cupriavidus pinatubonensis JMP134 involved in 2-hydroxyphenylacetic acid catabolism, via homogentisate pathway Type
Year 2021 Publication Microbial Biotechnology Abbreviated Journal Microb. Biotechnol.
Volume 14 Issue 5 Pages 1944-1960
Abstract The self-sufficient cytochrome P450 RhF and its homologues belonging to the CYP116B subfamily have attracted considerable attention due to the potential for biotechnological applications based in their ability to catalyse an array of challenging oxidative reactions without requiring additional protein partners. In this work, we showed for the first time that a CYP116B self-sufficient cytochrome P450 encoded by the ohpA gene harboured by Cupriavidus pinatubonensis JMP134, a beta-proteobacterium model for biodegradative pathways, catalyses the conversion of 2-hydroxyphenylacetic acid (2-HPA) into homogentisate. Mutational analysis and HPLC metabolite detection in strain JMP134 showed that 2-HPA is degraded through the well-known homogentisate pathway requiring a 2-HPA 5-hydroxylase activity provided by OhpA, which was additionally supported by heterologous expression and enzyme assays. The ohpA gene belongs to an operon including also ohpT, coding for a substrate-binding subunit of a putative transporter, whose expression is driven by an inducible promoter responsive to 2-HPA in presence of a predicted OhpR transcriptional regulator. OhpA homologues can be found in several genera belonging to Actinobacteria and alpha-, beta- and gamma-proteobacteria lineages indicating a widespread distribution of 2-HPA catabolism via homogentisate route. These results provide first time evidence for the natural function of members of the CYP116B self-sufficient oxygenases and represent a significant input to support novel kinetic and structural studies to develop cytochrome P450-based biocatalytic processes.
Corporate Author Thesis
Publisher Place of Publication Editor
Language Summary Language Original Title
Series Editor Series Title Abbreviated Series Title
Series Volume Series Issue Edition
ISSN 1751-7915 ISBN Medium
Area Expedition Conference
Notes WOS:000664134700001 Approved
Call Number UAI @ alexi.delcanto @ Serial 1426
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Author (up) Zuniga, A.; de la Fuente, F.; Federici, F.; Lionne, C.; Bonnet, J.; de Lorenzo, V.; Gonzalez, B.
Title An Engineered Device for Indoleacetic Acid Production under Quorum Sensing Signals Enables Cupriavidus pinatubonensis JMP134 To Stimulate Plant Growth Type
Year 2018 Publication Acs Synthetic Biology Abbreviated Journal ACS Synth. Biol.
Volume 7 Issue 6 Pages 1519-1527
Keywords Arabidopsis thaliana; Cupriavidus pinatubonensis; quorum sensing; synthetic beneficial interaction
Abstract The environmental effects of chemical fertilizers and pesticides have encouraged the quest for new strategies to increase crop productivity with minimal impacts on the natural medium. Plant growth promoting rhizobacteria (PGPR) can contribute to this endeavor by improving fitness through better nutrition acquisition and stress tolerance. Using the neutral (non PGPR) rhizobacterium Cupriavidus pinatubonensis JMP134 as the host, we engineered a regulatory forward loop that triggered the synthesis of the phytohormone indole-3-acetic acid (IAA) in a manner dependent on quorum sensing (QS) signals. Implementation of the device in JMP134 yielded synthesis of IAA in an autoregulated manner, improving the growth of the roots of inoculated Arabidopsis thaliana. These results not only demonstrated the value of the designed genetic module, but also validated C. pinatubonensis JMP134 as a suitable vehicle for agricultural applications, as it is amenable to genetic manipulations.
Address [Zuniga, Ana; de la Fuente, Francisco; Gonzalez, Bernardo] Univ Adolfo Ibanez, Fac Ingn & Ciencias, Ctr Appl Ecol & Sustainabil, Santiago 2640, Chile, Email: ana.zuniga@cbs.cnrs.fr
Corporate Author Thesis
Publisher Amer Chemical Soc Place of Publication Editor
Language English Summary Language Original Title
Series Editor Series Title Abbreviated Series Title
Series Volume Series Issue Edition
ISSN 2161-5063 ISBN Medium
Area Expedition Conference
Notes WOS:000435746400005 Approved
Call Number UAI @ eduardo.moreno @ Serial 877
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