Improved production of propionic acid using genome shuffling.
Traditionally derived from fossil fuels, biological production of propionic acid has recently gained interest. Propionibacterium species produce propionic acid as their main fermentation product. Production of other organic acids reduces propionic acid yield and productivity, pointing to by-products gene-knockout strategies as a logical solution to increase yield. However, removing by-product formation has seen limited success due to our inability to genetically engineer the best producing strains (i.e. Propionibacterium acidipropionici). To overcome this limitation, random mutagenesis continues to be the best path towards improving strains for biological propionic acid production. Recent advances in next generation sequencing opened new avenues to understand improved strains. In this work, we use genome shuffling on two wild type strains to generate a better propionic acid producing strain. Using next generation sequencing, we mapped the genomic changes leading to the improved phenotype. The best strain produced 25% more propionic acid than the wild type strain. Sequencing of the strains showed that genomic changes were restricted to single point mutations and gene duplications in well-conserved regions in the genomes. Such results confirm the involvement of gene conversion in genome shuffling as opposed to long genomic insertions.
Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Brisbane, Qld, Australia.
Carlos H
CH
Luna-Flores
Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Brisbane, Qld, Australia.
Robin W
RW
Palfreyman
Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Brisbane, Qld, Australia.
Jens O
JO
Krömer
Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Brisbane, Qld, Australia.
Lars K
LK
Nielsen
Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Brisbane, Qld, Australia.|Dow Centre for Sustainable Engineering and Innovation, The University of Queensland, Brisbane, Qld, Australia.
Esteban
E
Marcellin
D001709:Biotechnology / Q000379:methods*; D032621:DNA Shuffling; D059014:High-Throughput Nucleotide Sequencing; D011422:Propionates / Q000378:metabolism*; D011424:Propionibacterium / Q000235:genetics / Q000378:metabolism
D011422:Propionates; C029658:propionic acid
Genome shuffling; Next generation sequencing; Propionibacterium acidipropionici; Propionic acid; Protoplast fusion