
ABSTRACT Pseudomonas putida is a saprophytic bacterium with robust metabolisms and strong solvent tolerance making it an attractive host for metabolic engineering and bioremediation. Due to its diverse carbon metabolisms, its genome encodes an array of proteins and enzymes that can be readily applied to produce valuable products. In this work we sought to identify design principles and bottlenecks in the production of type III polyketide synthase (T3PKS)-derived compounds in P. putida . T3PKS products are widely used as nutraceuticals and medicines and often require aromatic starter units, such as coumaroyl-CoA, which is also an intermediate in the native coumarate catabolic pathway of P. putida . Using a randomly barcoded transposon mutant (RB-TnSeq) library, we assayed gene functions for a large portion of aromatic catabolism, confirmed known pathways, and proposed new annotations for two aromatic transporters. The 1,3,6,8-tetrahydroxynapthalene synthase of Streptomyces coelicolor (RppA), a microbial T3PKS, was then used to rapidly assay growth conditions for increased T3PKS product accumulation. The feruloyl/coumaroyl CoA synthetase (Fcs) of P. putida was used to supply coumaroyl-CoA for the curcuminoid synthase (CUS) of Oryza sativa , a plant T3PKS. We identified that accumulation of coumaroyl-CoA in this pathway results in extended growth lag times in P. putida . Deletion of the second step in coumarate catabolism, the enoyl-CoA hydratase-lyase (Ech), resulted in increased production of the type III polyketide bisdemethoxycurcumin.
570, 31 Biological Sciences (for-2020), QH301-705.5, 610, Biological Sciences, Industrial Biotechnology, Infectious Diseases, 3106 Industrial Biotechnology (for-2020), Infectious Diseases (rcdc), Special issue on Non-conventional microbes edited by Ian Wheeldon and Aindrila Mukhopadhyay, Biochemistry and cell biology, Medical biochemistry and metabolomics, Biology (General), 3101 Biochemistry and cell biology (for-2020), TP248.13-248.65, 3205 Medical biochemistry and metabolomics (for-2020), Biotechnology
570, 31 Biological Sciences (for-2020), QH301-705.5, 610, Biological Sciences, Industrial Biotechnology, Infectious Diseases, 3106 Industrial Biotechnology (for-2020), Infectious Diseases (rcdc), Special issue on Non-conventional microbes edited by Ian Wheeldon and Aindrila Mukhopadhyay, Biochemistry and cell biology, Medical biochemistry and metabolomics, Biology (General), 3101 Biochemistry and cell biology (for-2020), TP248.13-248.65, 3205 Medical biochemistry and metabolomics (for-2020), Biotechnology
| selected citations These citations are derived from selected sources. This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically). | 59 | |
| popularity This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network. | Top 1% | |
| influence This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically). | Top 10% | |
| impulse This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network. | Top 10% |
