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Title: Bioconversion of methane to lactate by an obligate methanotrophic bacterium

Abstract

Methane is the second most abundant greenhouse gas (GHG), with nearly 60% of emissions derived from anthropogenic sources. Microbial conversion of methane to fuels and value-added chemicals offers a means to reduce GHG emissions, while also valorizing this otherwise squandered high-volume, high-energy gas. However, to date, advances in methane biocatalysis have been constrained by the low-productivity and limited genetic tractability of natural methane-consuming microbes. Here, leveraging recent identification of a novel, tractable methanotrophic bacterium, Methylomicrobium buryatense, we demonstrate microbial biocatalysis of methane to lactate, an industrial platform chemical. Heterologous overexpression of a Lactobacillus helveticus L-lactate dehydrogenase in M. buryatense resulted in an initial titer of 0.06 g lactate/L from methane. Cultivation in a 5 L continuously stirred tank bioreactor enabled production of 0.8 g lactate/L, representing a 13-fold improvement compared to the initial titer. The yields (0.05 g lactate/g methane) and productivity (0.008 g lactate/L/h) indicate the need and opportunity for future strain improvement. Additionally, real-time analysis of methane utilization implicated gas-to-liquid transfer and/or microbial methane consumption as process limitations. This work opens the door to develop an array of methanotrophic bacterial strain-engineering strategies currently employed for biocatalytic sugar upgrading to “green” chemicals and fuels.

Authors:
 [1];  [1];  [1];  [2];  [1];  [1]
  1. National Renewable Energy Lab. (NREL), Golden, CO (United States)
  2. San Diego State Univ., San Diego, CA (United States)
Publication Date:
Research Org.:
National Renewable Energy Laboratory (NREL), Golden, CO (United States)
Sponsoring Org.:
USDOE Office of Energy Efficiency and Renewable Energy (EERE), Sustainable Transportation Office. Bioenergy Technologies Office
OSTI Identifier:
1239888
Report Number(s):
NREL/JA-5100-65849
Journal ID: ISSN 2045-2322
Grant/Contract Number:  
AC36-08GO28308
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Scientific Reports
Additional Journal Information:
Journal Volume: 6; Related Information: Scientific Reports; Journal ID: ISSN 2045-2322
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL, AND ANAYLYTICAL CHEMISTRY; methane; greenhouse gas (GHG); microbial biocatalysis; lactate; applied microbiology; metabolic engineering

Citation Formats

Henard, Calvin A., Smith, Holly, Dowe, Nancy, Kalyuzhnaya, Marina G., Pienkos, Philip T., and Guarnieri, Michael T. Bioconversion of methane to lactate by an obligate methanotrophic bacterium. United States: N. p., 2016. Web. doi:10.1038/srep21585.
Henard, Calvin A., Smith, Holly, Dowe, Nancy, Kalyuzhnaya, Marina G., Pienkos, Philip T., & Guarnieri, Michael T. Bioconversion of methane to lactate by an obligate methanotrophic bacterium. United States. https://doi.org/10.1038/srep21585
Henard, Calvin A., Smith, Holly, Dowe, Nancy, Kalyuzhnaya, Marina G., Pienkos, Philip T., and Guarnieri, Michael T. 2016. "Bioconversion of methane to lactate by an obligate methanotrophic bacterium". United States. https://doi.org/10.1038/srep21585. https://www.osti.gov/servlets/purl/1239888.
@article{osti_1239888,
title = {Bioconversion of methane to lactate by an obligate methanotrophic bacterium},
author = {Henard, Calvin A. and Smith, Holly and Dowe, Nancy and Kalyuzhnaya, Marina G. and Pienkos, Philip T. and Guarnieri, Michael T.},
abstractNote = {Methane is the second most abundant greenhouse gas (GHG), with nearly 60% of emissions derived from anthropogenic sources. Microbial conversion of methane to fuels and value-added chemicals offers a means to reduce GHG emissions, while also valorizing this otherwise squandered high-volume, high-energy gas. However, to date, advances in methane biocatalysis have been constrained by the low-productivity and limited genetic tractability of natural methane-consuming microbes. Here, leveraging recent identification of a novel, tractable methanotrophic bacterium, Methylomicrobium buryatense, we demonstrate microbial biocatalysis of methane to lactate, an industrial platform chemical. Heterologous overexpression of a Lactobacillus helveticus L-lactate dehydrogenase in M. buryatense resulted in an initial titer of 0.06 g lactate/L from methane. Cultivation in a 5 L continuously stirred tank bioreactor enabled production of 0.8 g lactate/L, representing a 13-fold improvement compared to the initial titer. The yields (0.05 g lactate/g methane) and productivity (0.008 g lactate/L/h) indicate the need and opportunity for future strain improvement. Additionally, real-time analysis of methane utilization implicated gas-to-liquid transfer and/or microbial methane consumption as process limitations. This work opens the door to develop an array of methanotrophic bacterial strain-engineering strategies currently employed for biocatalytic sugar upgrading to “green” chemicals and fuels.},
doi = {10.1038/srep21585},
url = {https://www.osti.gov/biblio/1239888}, journal = {Scientific Reports},
issn = {2045-2322},
number = ,
volume = 6,
place = {United States},
year = {Tue Feb 23 00:00:00 EST 2016},
month = {Tue Feb 23 00:00:00 EST 2016}
}

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Works referenced in this record:

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Expression of bacterial hemoglobin genes to improve astaxanthin production in a methanotrophic bacterium Methylomonas sp.
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Construction of the astaxanthin biosynthetic pathway in a methanotrophic bacterium Methylomonas sp. strain 16a
journal, January 2007


High-rate conversion of methane to methanol by Methylosinus trichosporium OB3b
journal, August 2011


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Engineering and adaptive evolution of Escherichia coli for d-lactate fermentation reveals GatC as a xylose transporter
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journal, May 2015


Metabolic engineering of Yarrowia lipolytica for itaconic acid production
journal, November 2015


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journal, August 2010


Identification and microbial production of a terpene-based advanced biofuel
journal, September 2011


Highly efficient methane biocatalysis revealed in a methanotrophic bacterium
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Control of Redox Balance by the Stringent Response Regulatory Protein Promotes Antioxidant Defenses of Salmonella
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L-lactic acid production from D-xylose with Candida sonorensis expressing a heterologous lactate dehydrogenase encoding gene
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Works referencing / citing this record:

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Biological conversion of methane to chemicals and fuels: technical challenges and issues
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A modular approach for high-flux lactic acid production from methane in an industrial medium using engineered Methylomicrobium buryatense 5GB1
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Methane utilization in Methylomicrobium alcaliphilum 20ZR: a systems approach
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Muconic acid production from methane using rationally-engineered methanotrophic biocatalysts
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Methanol-based acetoin production by genetically engineered Bacillus methanolicus
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Industrial biomanufacturing: The future of chemical production
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Development of a CRISPR/Cas9 System for Methylococcus capsulatus In Vivo Gene Editing
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Role of the malic enzyme in metabolism of the halotolerant methanotroph Methylotuvimicrobium alcaliphilum 20Z
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Fatty Acid Biosynthesis Pathways in Methylomicrobium buryatense 5G(B1)
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Oxygen-limited metabolism in the methanotroph Methylomicrobium buryatense 5GB1C
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Development of a CRISPR/Cas9 System for Methylococcus capsulatus In Vivo Gene Editing
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Fatty Acid Biosynthesis Pathways in Methylomicrobium buryatense 5G(B1)
journal, January 2017


Oxygen-limited metabolism in the methanotroph Methylomicrobium buryatense 5GB1C
journal, January 2017