Alkane biosynthesis by Aspergillus carbonarius ITEM 5010 through heterologous expression of Synechococcus elongatus acyl-ACP/CoA reductase and aldehyde deformylating oxygenase genes
Abstract
In this study we describe the heterologous expression of the recently identified cyanobacterial pathway for long chain alkane biosynthesis, involving the reduction of fatty acyl-ACP to fatty aldehyde and the subsequent conversion of this into alkanes, in the filamentous fungus Aspergillus carbonarius ITEM 5010. Genes originating from Synechococcus elongatus strain PCC7942, encoding acyl-ACP/CoA reductase and aldehyde deformylating oxygenase enzymes, were successfully expressed in A. carbonarius, which lead to the production of pentadecane and heptadecane, alkanes that have not been previously produced by this fungus. Titers of 0.2, 0.5 and 2.7 mg/l pentadecane and 0.8, 1.6 and 10.2 mg/l heptadecane were achieved using glucose, Yeast malt and oatmeal media, respectively. Besides producing alkanes, we found elevated levels of internal free fatty acids and triglycerides in the alkane producing transformant. These findings can indicate that a yet unidentified, native fatty aldehyde dehydrogenase channels back the fatty aldehydes into the fatty acid metabolism, thus competing for substrate with the heterologously expressed fatty aldehyde deformylating oxygenase. These findings will potentially facilitate the future application of robust, fungal cell factories for the production of advanced biofuels from various substrates.
- Authors:
-
- Washington State Univ., Pullman, WA (United States). Sciences and Engineering Lab. Bioproducts
- Aalborg Univ. Copenhagen (Denmark). Section for Sustainable Biotechnology
- Washington State Univ., Pullman, WA (United States). Sciences and Engineering Lab. Bioproducts; Aalborg Univ. Copenhagen (Denmark). Section for Sustainable Biotechnology
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Chemical and Biological Process Development Group
- Publication Date:
- Research Org.:
- Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC)
- OSTI Identifier:
- 1627036
- Grant/Contract Number:
- AC05-76RL01830
- Resource Type:
- Accepted Manuscript
- Journal Name:
- AMB Express
- Additional Journal Information:
- Journal Volume: 7; Journal Issue: 1; Journal ID: ISSN 2191-0855
- Publisher:
- Springer
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; Biotechnology & Applied Microbiology; Advanced biofuels; Alkane biosynthesis; Aspergillus carbonarius ITEM 5010; Fungal transformation
Citation Formats
Sinha, Malavika, Weyda, István, Sørensen, Annette, Bruno, Kenneth S., and Ahring, Birgitte K. Alkane biosynthesis by Aspergillus carbonarius ITEM 5010 through heterologous expression of Synechococcus elongatus acyl-ACP/CoA reductase and aldehyde deformylating oxygenase genes. United States: N. p., 2017.
Web. doi:10.1186/s13568-016-0321-x.
Sinha, Malavika, Weyda, István, Sørensen, Annette, Bruno, Kenneth S., & Ahring, Birgitte K. Alkane biosynthesis by Aspergillus carbonarius ITEM 5010 through heterologous expression of Synechococcus elongatus acyl-ACP/CoA reductase and aldehyde deformylating oxygenase genes. United States. https://doi.org/10.1186/s13568-016-0321-x
Sinha, Malavika, Weyda, István, Sørensen, Annette, Bruno, Kenneth S., and Ahring, Birgitte K. Thu .
"Alkane biosynthesis by Aspergillus carbonarius ITEM 5010 through heterologous expression of Synechococcus elongatus acyl-ACP/CoA reductase and aldehyde deformylating oxygenase genes". United States. https://doi.org/10.1186/s13568-016-0321-x. https://www.osti.gov/servlets/purl/1627036.
@article{osti_1627036,
title = {Alkane biosynthesis by Aspergillus carbonarius ITEM 5010 through heterologous expression of Synechococcus elongatus acyl-ACP/CoA reductase and aldehyde deformylating oxygenase genes},
author = {Sinha, Malavika and Weyda, István and Sørensen, Annette and Bruno, Kenneth S. and Ahring, Birgitte K.},
abstractNote = {In this study we describe the heterologous expression of the recently identified cyanobacterial pathway for long chain alkane biosynthesis, involving the reduction of fatty acyl-ACP to fatty aldehyde and the subsequent conversion of this into alkanes, in the filamentous fungus Aspergillus carbonarius ITEM 5010. Genes originating from Synechococcus elongatus strain PCC7942, encoding acyl-ACP/CoA reductase and aldehyde deformylating oxygenase enzymes, were successfully expressed in A. carbonarius, which lead to the production of pentadecane and heptadecane, alkanes that have not been previously produced by this fungus. Titers of 0.2, 0.5 and 2.7 mg/l pentadecane and 0.8, 1.6 and 10.2 mg/l heptadecane were achieved using glucose, Yeast malt and oatmeal media, respectively. Besides producing alkanes, we found elevated levels of internal free fatty acids and triglycerides in the alkane producing transformant. These findings can indicate that a yet unidentified, native fatty aldehyde dehydrogenase channels back the fatty aldehydes into the fatty acid metabolism, thus competing for substrate with the heterologously expressed fatty aldehyde deformylating oxygenase. These findings will potentially facilitate the future application of robust, fungal cell factories for the production of advanced biofuels from various substrates.},
doi = {10.1186/s13568-016-0321-x},
journal = {AMB Express},
number = 1,
volume = 7,
place = {United States},
year = {Thu Jan 05 00:00:00 EST 2017},
month = {Thu Jan 05 00:00:00 EST 2017}
}
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Point mutation of the xylose reductase (XR) gene reduces xylitol accumulation and increases citric acid production in Aspergillus carbonarius
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- Journal of Industrial Microbiology & Biotechnology, Vol. 41, Issue 4
Effects of heterologous expression of phosphoenolpyruvate carboxykinase and phosphoenolpyruvate carboxylase on organic acid production in Aspergillus carbonarius
journal, September 2015
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- Journal of Industrial Microbiology & Biotechnology, Vol. 42, Issue 11
Fatty aldehyde and fatty alcohol metabolism: Review and importance for epidermal structure and function
journal, March 2014
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- Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, Vol. 1841, Issue 3
Production of hydrocarbon compounds by endophytic fungi Gliocladium species grown on cellulose
journal, October 2011
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Production of hydrocarbons by Aspergillus carbonarius ITEM 5010
journal, April 2015
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Identification and characterization of the polyketide synthase involved in ochratoxin A biosynthesis in Aspergillus carbonarius
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A review on microbial synthesis of hydrocarbons
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Quantitative in Vivo Microsampling for Pharmacokinetic Studies Based on an Integrated Solid-Phase Microextraction System
journal, June 2007
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Evidence for Only Oxygenative Cleavage of Aldehydes to Alk(a/e)nes and Formate by Cyanobacterial Aldehyde Decarbonylases
journal, September 2012
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Production and characterization of β-glucosidases from different Aspergillus strains
journal, July 2001
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Taxonomy and significance of black aspergilli
journal, August 2004
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- Antonie van Leeuwenhoek, Vol. 86, Issue 1
Microbial engineering for the production of advanced biofuels
journal, August 2012
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Microbial production of short-chain alkanes
journal, September 2013
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Sjögren–Larsson syndrome is caused by mutations in the fatty aldehyde dehydrogenase gene
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Expanding metabolism for biosynthesis of nonnatural alcohols
journal, December 2008
- Zhang, K.; Sawaya, M. R.; Eisenberg, D. S.
- Proceedings of the National Academy of Sciences, Vol. 105, Issue 52
The production of myco-diesel hydrocarbons and their derivatives by the endophytic fungus Gliocladium roseum (NRRL 50072)
journal, November 2008
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- Microbiology, Vol. 154, Issue 11
Hydrocarbons of Blue-Green Algae: Geochemical Signfficance
journal, January 1969
- Winters, K.; Parker, P. L.; Van Baalen, C.
- Science, Vol. 163, Issue 3866
Deletion of glucose oxidase changes the pattern of organic acid production in Aspergillus carbonarius
journal, August 2014
- Yang, Lei; Lübeck, Mette; Lübeck, Peter S.
- AMB Express, Vol. 4, Issue 1
Fatty Aldehydes in Cyanobacteria Are a Metabolically Flexible Precursor for a Diversity of Biofuel Products
journal, March 2013
- Kaiser, Brett K.; Carleton, Michael; Hickman, Jason W.
- PLoS ONE, Vol. 8, Issue 3
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A direct method for fatty acid methyl ester synthesis: Application to wet meat tissues, oils, and feedstuffs
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- Journal of Animal Science, Vol. 85, Issue 6
Works referencing / citing this record:
The influence of fatty acid supply and aldehyde reductase deletion on cyanobacteria alkane generating pathway in Escherichia coli
journal, March 2018
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Identification of non-conserved residues essential for improving the hydrocarbon-producing activity of cyanobacterial aldehyde-deformylating oxygenase
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A novel C-terminal degron identified in bacterial aldehyde decarbonylases using directed evolution
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