Engineering Pseudomonas putida for isoprenoid production by manipulating endogenous and shunt pathways supplying precursors
Hernandez-Arranz, Sofia (Centre de Recerca en Agrigenòmica)
Pérez-Gil, Jordi (Centre de Recerca en Agrigenòmica)
Marshall-Sabey, D. (Centre de Recerca en Agrigenòmica)
Rodríguez Concepción, Manuel (Centre de Recerca en Agrigenòmica)

Date:	2019
Abstract:	Background: The soil bacterium Pseudomonas putida is a promising platform for the production of industrially valuable natural compounds. In the case of isoprenoids, the availability of biosynthetic precursors is a major limiting factor. In P. putida and most other bacteria, these precursors are produced from pyruvate and glyceraldehyde 3-phosphate by the methylerythritol 4-phosphate (MEP) pathway, whereas other bacteria synthesize the same precursors from acetyl-CoA using the unrelated mevalonate (MVA) pathway. Results: Here we explored different strategies to increase the supply of isoprenoid precursors in P. putida cells using lycopene as a read-out. Because we were not aiming at producing high isoprenoid titers but were primarily interested in finding ways to enhance the metabolic flux to isoprenoids, we engineered the well-characterized P. putida strain KT2440 to produce low but detectable levels of lycopene under conditions in which MEP pathway steps were not saturated. Then, we compared lycopene production in cells expressing the Myxococcus xanthus MVA pathway genes or endogenous MEP pathway genes (dxs, dxr, idi) under the control of IPTG-induced and stress-regulated promoters. We also tested a shunt pathway producing isoprenoid precursors from ribulose 5-phosphate using a mutant version of the Escherichia coli ribB gene. Conclusions: The most successful combination led to a 50-fold increase in lycopene levels, indicating that P. putida can be successfully engineered to substantially increase the supply of metabolic substrates for the production of industrially valuable isoprenoids.
Grants:	Agència de Gestió d'Ajuts Universitaris i de Recerca 2017/SGR‑710 European Commission 627639 Ministerio de Economía y Competitividad SEV‑2015‑0533 Ministerio de Ciencia e Innovación BIO2017‑90877‑REDT Ministerio de Ciencia e Innovación PCIN‑2015‑103
Rights:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Language:	Anglès
Document:	Article ; recerca ; Versió publicada
Published in:	Microbial cell factories, Vol. 18 (September 2019) , art. 152, ISSN 1475-2859

DOI: 10.1186/s12934-019-1204-z
PMID: 31500633

14 p, 1.7 MB

The record appears in these collections:
Research literature > UAB research groups literature > Research Centres and Groups (research output) > Experimental sciences > CRAG (Centre for Research in Agricultural Genomics)
Articles > Research articles
Articles > Published articles