Abstract
The biosynthesis of natural products by heterologous expression of biosynthetic pathways in amenable production strains enables biotechnological access to a variety of valuable compounds by conversion of renewable resources. Pseudomonas putida has emerged as a microbial laboratory work horse, with elaborated techniques for cultivation and genetic manipulation available. Beyond that, this bacterium offers several particular advantages with regard to natural product biosynthesis, notably a versatile intrinsic metabolism with diverse enzymatic capacities as well as an outstanding tolerance to xenobiotics. Therefore, it has been applied for recombinant biosynthesis of several valuable natural products. This review provides an overview of applications of P. putida as a host organism for the recombinant biosynthesis of such natural products, including rhamnolipids, terpenoids, polyketides and non-ribosomal peptides, and other amino acid-derived compounds. The focus is on de novo natural product synthesis from intrinsic building blocks by means of heterologous gene expression and strain engineering. Finally, the future potential of the bacterium as a chassis organism for synthetic microbiology is pointed out.
Highlights
For the biotechnological production of natural products, engineered bacteria generally offer several advantages over the original producers
As opposed to many natural producers, typically applied engineered bacteria are characterized by easy handling regarding laboratory cultivation which is the prerequisite for biotechnological applications
Usage of non-harmful generally recognized as safe (GRAS)-certified strains such as Pseudomonas putida KT2440 allows studies in many laboratories as well as industrial-scale production
Summary
For the biotechnological production of natural products, engineered bacteria generally offer several advantages over the original producers. T-cinnamate and phenol producer strains were generated by introduction of the phenylalanine/tyrosine ammonia lyase gene pal from Rhodosporidium toruloides ATCC 64815 and the tyrosine phenol lyase gene tpl from Pantoea agglomerans AJ2985, respectively (Nijkamp et al 2005; Wierckx et al 2005) Heterologous expression of these key biosynthetic enzymes was controlled by the salicylateinducible promoter system NagR/PnagAa from Comomonas testosteroni (tpl) or Ptac (pal) and resulted after further strain improvement by chemical mutagenesis in production strains that accumulated extracellularly 5.4 mM (0.8 g/l) t-cinnamate and 9.2 mM (0.9 g/l) phenol, respectively. Such studies demonstrate the potential of P. putida in highly diverse fields of application and may inspire further exciting developments towards the establishment of P. putida as a platform for production of various natural products in the future
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