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  • Overexpression of native carbonic anhydrases increases carbon conversion efficiency in the methanotrophic biocatalyst Methylococcus capsulatus Bath
Title: Overexpression of native carbonic anhydrases increases carbon conversion efficiency in the methanotrophic biocatalyst Methylococcus capsulatus Bath
ABSTRACT Methanotrophic bacteria play a vital role in the biogeochemical carbon cycle due to their unique ability to use CH4as a carbon and energy source. Evidence suggests that some methanotrophs, includingMethylococcus capsulatus, can also use CO2as a carbon source, making these bacteria promising candidates for developing biotechnologies targeting greenhouse gas capture and mitigation. However, a deeper understanding of the dual CH4and CO2metabolism is needed to guide methanotroph strain improvements and realize their industrial utility. In this study, we show thatM. capsulatusexpresses five carbonic anhydrase (CA) isoforms, one α-CA, one γ-CA, and three β-CAs, that play a role in its inorganic carbon metabolism and CO2-dependent growth. The CA isoforms are differentially expressed, and transcription of all isoform genes is induced in response to CO2limitation. CA null mutant strains exhibited markedly impaired growth compared to an isogenic wild-type control, suggesting that the CA isoforms have independent, non-redundant roles inM. capsulatusmetabolism and physiology. Overexpression of some, but not all, CA isoforms improved bacterial growth kinetics and decreased CO2evolution from CH4-consuming cultures. Notably, we developed an engineered methanotrophic biocatalyst overexpressing the native α-CA and β-CA with a 2.5-fold improvement in the conversion of CH4to biomass. Given that product yield is a significant cost driver of methanotroph-based bioprocesses, the engineered strain developed here could improve the economics of CH4biocatalysis, including the production of single-cell protein from natural gas or anaerobic digestion-derived biogas.IMPORTANCEMethanotrophs transform CH4into CO2and multi-carbon compounds, so they play a critical role in the global carbon cycle and are of interest for biotechnology applications. Some methanotrophs, includingMethylococcus capsulatus, can also use CO2as a carbon source, but this dual one-carbon metabolism is incompletely understood. In this study, we show thatM. capsulatuscarbonic anhydrases are critical for this bacterium to optimally utilize CO2. We developed an engineered strain with improved CO2utilization capacity that increased the overall carbon conversion to cell biomass. The improvements to methanotroph-based product yields observed here are expected to reduce costs associated with CH4conversion bioprocesses.  more » « less
Award ID(s):
2225776
PAR ID:
10536722
Author(s) / Creator(s):
; ; ; ; ;
Editor(s):
Tringe, Susannah Green
Publisher / Repository:
American Society for Microbiology
Date Published:
Journal Name:
mSphere
ISSN:
2379-5042
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
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