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:
-
- National Renewable Energy Lab. (NREL), Golden, CO (United States)
- 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}
}
Web of Science
Works referenced in this record:
Metabolic engineering in methanotrophic bacteria
journal, May 2015
- Kalyuzhnaya, Marina G.; Puri, Aaron W.; Lidstrom, Mary E.
- Metabolic Engineering, Vol. 29
Isolation and Characterization of Halotolerant Alkaliphilic Methanotrophic Bacteria from Tuva Soda Lakes
journal, November 1997
- Khmelenina, Valentina N.
- Current Microbiology, Vol. 35, Issue 5
Accurate and reliable quantification of total microalgal fuel potential as fatty acid methyl esters by in situ transesterification
journal, February 2012
- Laurens, Lieve M. L.; Quinn, Matthew; Van Wychen, Stefanie
- Analytical and Bioanalytical Chemistry, Vol. 403, Issue 1
Microbial production of poly(hydroxybutyrate) from C1 carbon sources
journal, January 2013
- Khosravi-Darani, Kianoush; Mokhtari, Zahra-Beigom; Amai, Tomohito
- Applied Microbiology and Biotechnology, Vol. 97, Issue 4
Draft Genome Sequence of Methylomicrobium buryatense Strain 5G, a Haloalkaline-Tolerant Methanotrophic Bacterium
journal, June 2013
- Khmelenina, V. N.; Beck, D. A. C.; Munk, C.
- Genome Announcements, Vol. 1, Issue 4
Envisioning the Bioconversion of Methane to Liquid Fuels
journal, February 2014
- Conrado, R. J.; Gonzalez, R.
- Science, Vol. 343, Issue 6171
Efficient Production of L-Lactic Acid by Metabolically Engineered Saccharomyces cerevisiae with a Genome-Integrated L-Lactate Dehydrogenase Gene
journal, April 2005
- Ishida, N.; Saitoh, S.; Tokuhiro, K.
- Applied and Environmental Microbiology, Vol. 71, Issue 4
Genetic Systems for Moderately Halo(alkali)philic Bacteria of the Genus Methylomicrobium
book, January 2011
- Ojala, David S.; Beck, David A. C.; Kalyuzhnaya, Marina G.
- Methods in Methane Metabolism, Part B: Methanotrophy
Taxonomic Characterization of New Alkaliphilic and Alkalitolerant Methanotrophs from Soda Lakes of the Southeastern Transbaikal Region and description of Methylomicrobium buryatense sp.nov.
journal, January 2001
- Kaluzhnaya, Marina; Khmelenina, Valentina; Eshinimaev, Bulat
- Systematic and Applied Microbiology, Vol. 24, Issue 2
Cow power: the energy and emissions benefits of converting manure to biogas
journal, July 2008
- Cuéllar, Amanda D.; Webber, Michael E.
- Environmental Research Letters, Vol. 3, Issue 3
Bioconversion of natural gas to liquid fuel: Opportunities and challenges
journal, May 2014
- Fei, Qiang; Guarnieri, Michael T.; Tao, Ling
- Biotechnology Advances, Vol. 32, Issue 3
Intracellular localization of the particulate methane monooxygenase and methanol dehydrogenase in Methylomicrobium album BG8
journal, July 2002
- Brantner, Christine; Remsen, Charles; Owen, Heather
- Archives of Microbiology, Vol. 178, Issue 1
Glycolytic enzymes in mammalian spermatozoa. Activities and stabilities of hexokinase and phosphofructokinase in various fractions from sperm homogenates
journal, October 1971
- Harrison, R. A. P.
- Biochemical Journal, Vol. 124, Issue 4
Rethinking biological activation of methane and conversion to liquid fuels
journal, April 2014
- Haynes, Chad A.; Gonzalez, Ramon
- Nature Chemical Biology, Vol. 10, Issue 5
Methane as a Resource: Can the Methanotrophs Add Value?
journal, March 2015
- Strong, P. J.; Xie, S.; Clarke, W. P.
- Environmental Science & Technology, Vol. 49, Issue 7
Heterotrophic bacteria growing in association with Methylococcus capsulatus (Bath) in a single cell protein production process
journal, June 2002
- Bothe, Harald; Jensen, K. Møller; A., Mergel
- Applied Microbiology and Biotechnology, Vol. 59, Issue 1
High-rate conversion of methane to methanol by Methylosinus trichosporium OB3b
journal, August 2011
- Duan, Changhong; Luo, Mingfang; Xing, Xinhui
- Bioresource Technology, Vol. 102, Issue 15
Biocatalytic Conversion of Methane to Methanol as a Key Step for Development of Methane-Based Biorefineries
journal, December 2014
- Hwang, In Yeub; Lee, Seung Hwan; Choi, Yoo Seong
- Journal of Microbiology and Biotechnology, Vol. 24, Issue 12
Wasted Food, Wasted Energy: The Embedded Energy in Food Waste in the United States
journal, August 2010
- Cuéllar, Amanda D.; Webber, Michael E.
- Environmental Science & Technology, Vol. 44, Issue 16
Expression of bacterial hemoglobin genes to improve astaxanthin production in a methanotrophic bacterium Methylomonas sp.
journal, November 2006
- Tao, Luan; Sedkova, Natalia; Yao, Henry
- Applied Microbiology and Biotechnology, Vol. 74, Issue 3
Identification and microbial production of a terpene-based advanced biofuel
journal, September 2011
- Peralta-Yahya, Pamela P.; Ouellet, Mario; Chan, Rossana
- Nature Communications, Vol. 2, Issue 1
Metabolic engineering of Yarrowia lipolytica for itaconic acid production
journal, November 2015
- Blazeck, John; Hill, Andrew; Jamoussi, Mariam
- Metabolic Engineering, Vol. 32
Recent advances in lactic acid production by microbial fermentation processes
journal, November 2013
- Abdel-Rahman, Mohamed Ali; Tashiro, Yukihiro; Sonomoto, Kenji
- Biotechnology Advances, Vol. 31, Issue 6
Use of Transposon Promoter-Probe Vectors in the Metabolic Engineering of the Obligate Methanotroph Methylomonas sp. Strain 16a for Enhanced C40 Carotenoid Synthesis
journal, January 2007
- Sharpe, P. L.; DiCosimo, D.; Bosak, M. D.
- Applied and Environmental Microbiology, Vol. 73, Issue 6
Microbial production of lactic acid
journal, January 2015
- Eiteman, Mark A.; Ramalingam, Subramanian
- Biotechnology Letters, Vol. 37, Issue 5
Use of the tetracycline promoter for the tightly regulated production of a murine antibody fragment in Escherichia coli
journal, December 1994
- Skerra, Arne
- Gene, Vol. 151, Issue 1-2
L-lactic acid production from D-xylose with Candida sonorensis expressing a heterologous lactate dehydrogenase encoding gene
journal, August 2014
- Koivuranta, Kari T.; Ilmén, Marja; Wiebe, Marilyn G.
- Microbial Cell Factories, Vol. 13, Issue 1
Lignin valorization through integrated biological funneling and chemical catalysis
journal, August 2014
- Linger, J. G.; Vardon, D. R.; Guarnieri, M. T.
- Proceedings of the National Academy of Sciences, Vol. 111, Issue 33, p. 12013-12018
Highly efficient methane biocatalysis revealed in a methanotrophic bacterium
journal, December 2013
- Kalyuzhnaya, M. G.; Yang, S.; Rozova, O. N.
- Nature Communications, Vol. 4, Issue 1
Engineering and adaptive evolution of Escherichia coli for d-lactate fermentation reveals GatC as a xylose transporter
journal, September 2012
- Utrilla, José; Licona-Cassani, Cuauhtemoc; Marcellin, Esteban
- Metabolic Engineering, Vol. 14, Issue 5
Genetic Tools for the Industrially Promising Methanotroph Methylomicrobium buryatense
journal, December 2014
- Puri, Aaron W.; Owen, Sarah; Chu, Frances
- Applied and Environmental Microbiology, Vol. 81, Issue 5
The Expanding World of Methylotrophic Metabolism
journal, October 2009
- Chistoserdova, Ludmila; Kalyuzhnaya, Marina G.; Lidstrom, Mary E.
- Annual Review of Microbiology, Vol. 63, Issue 1
Construction of the astaxanthin biosynthetic pathway in a methanotrophic bacterium Methylomonas sp. strain 16a
journal, January 2007
- Ye, Rick W.; Yao, Henry; Stead, Kristen
- Journal of Industrial Microbiology & Biotechnology, Vol. 34, Issue 4
Search for Methanotrophic Producers of Exopolysaccharides
journal, November 2001
- Malashenko, Yu. R.; Pirog, T. P.; Romanovskaya, V. A.
- Applied Biochemistry and Microbiology, Vol. 37, Issue 6, p. 599-602
Building carbon–carbon bonds using a biocatalytic methanol condensation cycle
journal, October 2014
- Bogorad, Igor W.; Chen, Chang-Ting; Theisen, Matthew K.
- Proceedings of the National Academy of Sciences, Vol. 111, Issue 45
Intracellular localization of the particulate methane monooxygenase and methanol dehydrogenase in Methylomicrobium album BG8
journal, July 2002
- Brantner, Christine; Remsen, Charles; Owen, Heather
- Archives of Microbiology, Vol. 178, Issue 1
Heterotrophic bacteria growing in association with Methylococcus capsulatus (Bath) in a single cell protein production process
journal, June 2002
- Bothe, Harald; Jensen, K. Møller; A., Mergel
- Applied Microbiology and Biotechnology, Vol. 59, Issue 1
Expression of bacterial hemoglobin genes to improve astaxanthin production in a methanotrophic bacterium Methylomonas sp.
journal, November 2006
- Tao, Luan; Sedkova, Natalia; Yao, Henry
- Applied Microbiology and Biotechnology, Vol. 74, Issue 3
Construction of the astaxanthin biosynthetic pathway in a methanotrophic bacterium Methylomonas sp. strain 16a
journal, January 2007
- Ye, Rick W.; Yao, Henry; Stead, Kristen
- Journal of Industrial Microbiology & Biotechnology, Vol. 34, Issue 4
Use of the tetracycline promoter for the tightly regulated production of a murine antibody fragment in Escherichia coli
journal, December 1994
- Skerra, Arne
- Gene, Vol. 151, Issue 1-2
High-rate conversion of methane to methanol by Methylosinus trichosporium OB3b
journal, August 2011
- Duan, Changhong; Luo, Mingfang; Xing, Xinhui
- Bioresource Technology, Vol. 102, Issue 15
Recent advances in lactic acid production by microbial fermentation processes
journal, November 2013
- Abdel-Rahman, Mohamed Ali; Tashiro, Yukihiro; Sonomoto, Kenji
- Biotechnology Advances, Vol. 31, Issue 6
Engineering and adaptive evolution of Escherichia coli for d-lactate fermentation reveals GatC as a xylose transporter
journal, September 2012
- Utrilla, José; Licona-Cassani, Cuauhtemoc; Marcellin, Esteban
- Metabolic Engineering, Vol. 14, Issue 5
Metabolic engineering in methanotrophic bacteria
journal, May 2015
- Kalyuzhnaya, Marina G.; Puri, Aaron W.; Lidstrom, Mary E.
- Metabolic Engineering, Vol. 29
Metabolic engineering of Yarrowia lipolytica for itaconic acid production
journal, November 2015
- Blazeck, John; Hill, Andrew; Jamoussi, Mariam
- Metabolic Engineering, Vol. 32
Wasted Food, Wasted Energy: The Embedded Energy in Food Waste in the United States
journal, August 2010
- Cuéllar, Amanda D.; Webber, Michael E.
- Environmental Science & Technology, Vol. 44, Issue 16
Identification and microbial production of a terpene-based advanced biofuel
journal, September 2011
- Peralta-Yahya, Pamela P.; Ouellet, Mario; Chan, Rossana
- Nature Communications, Vol. 2, Issue 1
Highly efficient methane biocatalysis revealed in a methanotrophic bacterium
journal, December 2013
- Kalyuzhnaya, M. G.; Yang, S.; Rozova, O. N.
- Nature Communications, Vol. 4, Issue 1
Control of Redox Balance by the Stringent Response Regulatory Protein Promotes Antioxidant Defenses of Salmonella
journal, September 2010
- Henard, Calvin A.; Bourret, Travis J.; Song, Miryoung
- Journal of Biological Chemistry, Vol. 285, Issue 47
Genome Sequence of the Haloalkaliphilic Methanotrophic Bacterium Methylomicrobium alcaliphilum 20Z
journal, December 2011
- Vuilleumier, S.; Khmelenina, V. N.; Bringel, F.
- Journal of Bacteriology, Vol. 194, Issue 2
L-lactic acid production from D-xylose with Candida sonorensis expressing a heterologous lactate dehydrogenase encoding gene
journal, August 2014
- Koivuranta, Kari T.; Ilmén, Marja; Wiebe, Marilyn G.
- Microbial Cell Factories, Vol. 13, Issue 1
Works referencing / citing this record:
Metabolic engineering of methanotrophs and its application to production of chemicals and biofuels from methane: Biofuel energy consumption-Economic growth relationship: An empirical investigation of Brazil
journal, August 2016
- Lee, Ok Kyung; Hur, Dong Hoon; Nguyen, Diep Thi Ngoc
- Biofuels, Bioproducts and Biorefining, Vol. 10, Issue 6
Building a genome engineering toolbox in nonmodel prokaryotic microbes
journal, May 2018
- Freed, Emily; Fenster, Jacob; Smolinski, Sharon L.
- Biotechnology and Bioengineering, Vol. 115, Issue 9
Biological conversion of methane to chemicals and fuels: technical challenges and issues
journal, February 2018
- Hwang, In Yeub; Nguyen, Anh Duc; Nguyen, Thu Thi
- Applied Microbiology and Biotechnology, Vol. 102, Issue 7
A modular approach for high-flux lactic acid production from methane in an industrial medium using engineered Methylomicrobium buryatense 5GB1
journal, April 2018
- Garg, Shivani; Clomburg, James M.; Gonzalez, Ramon
- Journal of Industrial Microbiology & Biotechnology, Vol. 45, Issue 6
Methane utilization in Methylomicrobium alcaliphilum 20ZR: a systems approach
journal, February 2018
- Akberdin, Ilya R.; Thompson, Merlin; Hamilton, Richard
- Scientific Reports, Vol. 8, Issue 1
Muconic acid production from methane using rationally-engineered methanotrophic biocatalysts
journal, January 2019
- Henard, Calvin A.; Akberdin, Ilya R.; Kalyuzhnaya, Marina G.
- Green Chemistry, Vol. 21, Issue 24
Methanol-based acetoin production by genetically engineered Bacillus methanolicus
journal, January 2020
- Drejer, Eivind B.; Chan, Dennis Tin Chat; Haupka, Carsten
- Green Chemistry, Vol. 22, Issue 3
Industrial biomanufacturing: The future of chemical production
journal, January 2017
- Clomburg, James M.; Crumbley, Anna M.; Gonzalez, Ramon
- Science, Vol. 355, Issue 6320
Development of a CRISPR/Cas9 System for Methylococcus capsulatus In Vivo Gene Editing
journal, March 2019
- Tapscott, Timothy; Guarnieri, Michael T.; Henard, Calvin A.
- Applied and Environmental Microbiology, Vol. 85, Issue 11
Biological conversion of methane to putrescine using genome-scale model-guided metabolic engineering of a methanotrophic bacterium Methylomicrobium alcaliphilum 20Z
journal, June 2019
- Nguyen, Linh Thanh; Lee, Eun Yeol
- Biotechnology for Biofuels, Vol. 12, Issue 1
Efficient production of d-lactate from methane in a lactate-tolerant strain of Methylomonas sp. DH-1 generated by adaptive laboratory evolution
journal, September 2019
- Lee, Jong Kwan; Kim, Sujin; Kim, Wonsik
- Biotechnology for Biofuels, Vol. 12, Issue 1
Role of the malic enzyme in metabolism of the halotolerant methanotroph Methylotuvimicrobium alcaliphilum 20Z
journal, November 2019
- Rozova, Olga N.; Mustakhimov, Ildar I.; But, Sergei Y.
- PLOS ONE, Vol. 14, Issue 11
Fatty Acid Biosynthesis Pathways in Methylomicrobium buryatense 5G(B1)
journal, January 2017
- Demidenko, Aleksandr; Akberdin, Ilya R.; Allemann, Marco
- Frontiers in Microbiology, Vol. 7
Oxygen-limited metabolism in the methanotroph Methylomicrobium buryatense 5GB1C
journal, January 2017
- Gilman, Alexey; Fu, Yanfen; Hendershott, Melissa
- PeerJ, Vol. 5
Two Experimental Protocols for Accurate Measurement of Gas Component Uptake and Production Rates in Bioconversion Processes
journal, April 2019
- Stone, Kyle A.; He, Q. Peter; Wang, Jin
- Scientific Reports, Vol. 9, Issue 1
Development of a CRISPR/Cas9 System for Methylococcus capsulatus In Vivo Gene Editing
journal, March 2019
- Tapscott, Timothy; Guarnieri, Michael T.; Henard, Calvin A.
- Applied and Environmental Microbiology, Vol. 85, Issue 11
Efficient production of d-lactate from methane in a lactate-tolerant strain of Methylomonas sp. DH-1 generated by adaptive laboratory evolution
journal, September 2019
- Lee, Jong Kwan; Kim, Sujin; Kim, Wonsik
- Biotechnology for Biofuels, Vol. 12, Issue 1
Fatty Acid Biosynthesis Pathways in Methylomicrobium buryatense 5G(B1)
journal, January 2017
- Demidenko, Aleksandr; Akberdin, Ilya R.; Allemann, Marco
- Frontiers in Microbiology, Vol. 7
A Prospective Study on the Fermentation Landscape of Gaseous Substrates to Biorenewables Using Methanosarcina acetivorans Metabolic Model
journal, August 2018
- Nazem-Bokaee, Hadi; Maranas, Costas D.
- Frontiers in Microbiology, Vol. 9
Oxygen-limited metabolism in the methanotroph Methylomicrobium buryatense 5GB1C
journal, January 2017
- Gilman, Alexey; Fu, Yanfen; Hendershott, Melissa
- PeerJ, Vol. 5