Pseudomonas as a microbial enzyme factory : the source of industrially potent enzymes and the host for heterologous enzyme production

The research presented in this thesis investigates, from different angles, Pseudomonas for the production of pharmaceutically relevant proteins. Pseudomonas can serve as a potential source of industrially potent enzymes and the host for nanotechnological production of biocatalysts. Therefore, Pseudomonads can be seen as microbial enzyme factories. In particular, the fundamental issues, such as the regulation of the lipase gene expression in P. alcaligenes, secretion of heterologous proteins and a search for industrially potent enzymes within P. aeruginosa, were studied here. As a result, we demonstrated at the molecular level that the LipRQ two-component system of P. alcaligenes governs the regulation of lipase expression by direct LipR binding to the lipase promoter. We also analyzed conditions inducing lipase expression. These findings are certainly important for the optimization of lipase production on industrial scale. Furthermore, the observations presented here, concerning PA1893, should stimulate future work extended by the list of PA1893 potential substrate compounds to begin with. Finally, a study with P. aeruginosa as a host for heterologous penicillin G acylase (PGA) expression and secretion suggests that for each and every individual heterologous protein to be expressed a number of different factors determines the production levels and the fate of the protein. It appears that the periplasmic space of P. aeruginosa can serve as a potent folding compartment for various proteins, yet achieving the successful secretion will require further studies before the somewhat universal secretion signal can be defined. Also, the efficient Pseudomonas factory might have to be, ultimately, engineered in a protein-specific manner to fulfill the requirements of a particular heterologous protein. Thus, it would be of great value to pursue on this broad and challenging research in near future to further identify the key factors important for protein expression and secretion within the Pseudomonas enzyme factories

	Title and contents
	Chapter 1
	Chapter 2
	Chapter 3
	Chapter 4
	Chapter 5
	Chapter 6
	Summary, general discussion
	Samenvatting
	Acknowledgements
	Complete thesis

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