The overarching aim of the project is to re-programme the lifestyle of Pseudomonas putida and designing a modular, streamlined bacterial platform for bespoke biocatalysis. Steps that lead to this overall aim are:
Step 1 - Generation of strains with a genome streamlined chassis still enabling growth and production but freed of non-essential functionalities and re-factored for optimal functioning.
Step 2 - Development of novel ATP sensoring and control loops.
Step 3 - Design and implementation of strains uncoupling growth from production of specialty and bulk chemicals.
Step 4 - Harness of non-native butanol and isobutanol production in re-factored chassis.
Step 5 - Opening the door for novel bio-sustainable routes by accessing butene and butadiene as well as a specialty chemical with potential use in crop protection from glucose.
Step 6 - Validation of the value and strength of the refactored Pseudomonas putida–based platform for the production of the bulk and specialty chemicals indicated above.
Step 7 - Translation of the knowledge and performance testing for potential end use applications.
This novel production platform shall comprise:
- a streamlined genome still enabling growth but liberated from non-essential functionalities and, presumably, more amenable for process control
- novel plug-in loci under the control of orthogonal regulation systems (small RNAs)
- novel ATP sensoring and control loops decoupling ATP availability from growth, and
- new pathways to produce ATP-demanding (iso-)butanol and derivatives as building blocks for chemicals and biofuels, as an example of a class of specialty chemical of high agricultural and industrial importance