Protoplast Fusion with the Hydrogen-Oxidizing Bacterium, Cupriavidus Necator

The hydrogen-oxidizing bacteria, Cupriavidus necator, is prominently known for its ability to naturally produce the biodegradable plastic, polyhydroxybutyrate (PHB).  Attempts to increase production of PHB have led to a number of tools enabling the bacteria’s genetic tractability.  A shortcoming of these tools is the ability to holistically modify C. necator’s genome.  A technique for this purpose which has been applied to other organisms is called protoplast fusion.  Protoplast fusion involves the removal of the bacteria's outermembrane and subsequent fusion of the cells allowing the genes of the various genomes to be shuffled.  To enable the use of protoplast fusion with C. necator, we developed a protocol and show the effectiveness of this technique using two strains of amino acid auxotrophs.  To date, this is the first application of this technique with C. necator.