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Title: Genetic Tools for the Industrially Promising Methanotroph Methylomicrobium buryatense

Abstract

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.

Authors:
; ; ; ; ; ;
Publication Date:
Sponsoring Org.:
USDOE Advanced Research Projects Agency - Energy (ARPA-E)
OSTI Identifier:
1211474
DOE Contract Number:  
DE-AR0000350
Resource Type:
Journal Article
Journal Name:
Applied and Environmental Microbiology
Additional Journal Information:
Journal Volume: 81; Journal Issue: 5; Journal ID: ISSN 0099-2240
Country of Publication:
United States
Language:
English

Citation Formats

Puri, AW, Owen, S, Chu, F, Chavkin, T, Beck, DAC, Kalyuzhnaya, MG, and Lidstrom, ME. Genetic Tools for the Industrially Promising Methanotroph Methylomicrobium buryatense. United States: N. p., 2015. Web. doi:10.1128/AEM.03795-14.
Puri, AW, Owen, S, Chu, F, Chavkin, T, Beck, DAC, Kalyuzhnaya, MG, & Lidstrom, ME. Genetic Tools for the Industrially Promising Methanotroph Methylomicrobium buryatense. United States. doi:10.1128/AEM.03795-14.
Puri, AW, Owen, S, Chu, F, Chavkin, T, Beck, DAC, Kalyuzhnaya, MG, and Lidstrom, ME. Tue . "Genetic Tools for the Industrially Promising Methanotroph Methylomicrobium buryatense". United States. doi:10.1128/AEM.03795-14.
@article{osti_1211474,
title = {Genetic Tools for the Industrially Promising Methanotroph Methylomicrobium buryatense},
author = {Puri, AW and Owen, S and Chu, F and Chavkin, T and Beck, DAC and Kalyuzhnaya, MG and Lidstrom, ME},
abstractNote = {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.},
doi = {10.1128/AEM.03795-14},
journal = {Applied and Environmental Microbiology},
issn = {0099-2240},
number = 5,
volume = 81,
place = {United States},
year = {2015},
month = {2}
}