Genetic Tools for the Industrially Promising Methanotroph Methylomicrobium buryatense

Journal Article · Tue Feb 10 04:00:00 EST 2015 · Applied and Environmental Microbiology

DOI:https://doi.org/10.1128/AEM.03795-14· OSTI ID:1211474

Puri, AW; Owen, S; Chu, F; Chavkin, T; Beck, DAC; Kalyuzhnaya, MG; Lidstrom, ME

Aerobic methanotrophs oxidize methane at ambient temperatures and pressures and are therefore attractive systems for methane-based bioconversions. In this work, we developed and validated genetic tools for Methylomicrobium buryatense, a haloalkaliphilic gammaproteobacterial (type I) methanotroph. M. buryatense was isolated directly on natural gas and grows robustly in pure culture with a 3-h doubling time, enabling rapid genetic manipulation compared to many other methanotrophic species. As a proof of concept, we used a sucrose counterselection system to eliminate glycogen production in M. buryatense by constructing unmarked deletions in two redundant glycogen synthase genes. We also selected for a more genetically tractable variant strain that can be conjugated with small incompatibility group P (IncP)-based broad-host-range vectors and determined that this capability is due to loss of the native plasmid. These tools make M. buryatense a promising model system for studying aerobic methanotroph physiology and enable metabolic engineering in this bacterium for industrial biocatalysis of methane.

• Sponsoring Organization: USDOE Advanced Research Projects Agency - Energy (ARPA-E)
• DOE Contract Number: AR0000350
• OSTI ID: 1211474
• Journal Information: Applied and Environmental Microbiology, Journal Name: Applied and Environmental Microbiology Journal Issue: 5 Vol. 81; ISSN 0099-2240
• Country of Publication: United States
• Language: English