Modulation of medium-chain fatty acid synthesis in Synechococcus sp. PCC 7002 by replacing FabH with a Chaetoceros Ketoacyl-ACP synthase
Abstract
The isolation or engineering of algal cells synthesizing high levels of medium-chain fatty acids (MCFAs) is attractive to mitigate the high clouding point of longer chain fatty acids in algal based biodiesel. To develop a more informed understanding of MCFA synthesis in photosynthetic microorganisms, we isolated several algae from Great Salt Lake and screened this collection for MCFA accumulation to identify strains naturally accumulating high levels of MCFA. A diatom, Chaetoceros sp. GSL56, accumulated particularly high levels of C14 (up to 40%), with the majority of C14 fatty acids allocated in triacylglycerols. Using whole cell transcriptome sequencing and de novo assembly, putative genes encoding fatty acid synthesis enzymes were identified. Enzymes from this Chaetoceros sp. were expressed in the cyanobacterium Synechococcus sp. PCC 7002 to validate gene function and to determine whether eukaryotic enzymes putatively lacking bacterial evolutionary control mechanisms could be used to improve MCFA production in this promising production strain. Replacement of the Synechococcus 7002 native FabH with a Chaetoceros ketoacyl-ACP synthase Ill increased MCFA synthesis up to fivefold. In conclusion, the level of increase is dependent on promoter strength and culturing conditions.
- Authors:
-
- Colorado School of Mines, Golden, CO, (United States). Dept. of Chemistry and Geochemistry
- Carnegie Institution for Science, Stanford, CA, (United States). Dept. of Plant Biology
- Publication Date:
- Research Org.:
- Colorado School of Mines, Golden, CO, (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22), Chemical Sciences, Geosciences and Biosciences Division
- OSTI Identifier:
- 1281962
- Grant/Contract Number:
- SC0008547
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Frontiers in Plant Science
- Additional Journal Information:
- Journal Volume: 7; Journal ID: ISSN 1664-462X
- Publisher:
- Frontiers Research Foundation
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; medium chain fatty acids; Chaetoceros; type III ketoacyl-ACP synthase; Synechococcus sp PCC 7002; lauric acid production; RNAseq; chlamydomonas-reinhardtii; biofuel production; lipid-composition; triacylglycerol accumulation; phaeodactylum-tricornutum; nitrogen starvation; escherichia-coli; fuel properties; life-cycle; microalgae
Citation Formats
Gu, Huiya, Jinkerson, Robert E., Davies, Fiona K., Sisson, Lyle A., Schneider, Philip E., and Posewitz, Matthew C. Modulation of medium-chain fatty acid synthesis in Synechococcus sp. PCC 7002 by replacing FabH with a Chaetoceros Ketoacyl-ACP synthase. United States: N. p., 2016.
Web. doi:10.3389/fpls.2016.00690.
Gu, Huiya, Jinkerson, Robert E., Davies, Fiona K., Sisson, Lyle A., Schneider, Philip E., & Posewitz, Matthew C. Modulation of medium-chain fatty acid synthesis in Synechococcus sp. PCC 7002 by replacing FabH with a Chaetoceros Ketoacyl-ACP synthase. United States. https://doi.org/10.3389/fpls.2016.00690
Gu, Huiya, Jinkerson, Robert E., Davies, Fiona K., Sisson, Lyle A., Schneider, Philip E., and Posewitz, Matthew C. Thu .
"Modulation of medium-chain fatty acid synthesis in Synechococcus sp. PCC 7002 by replacing FabH with a Chaetoceros Ketoacyl-ACP synthase". United States. https://doi.org/10.3389/fpls.2016.00690. https://www.osti.gov/servlets/purl/1281962.
@article{osti_1281962,
title = {Modulation of medium-chain fatty acid synthesis in Synechococcus sp. PCC 7002 by replacing FabH with a Chaetoceros Ketoacyl-ACP synthase},
author = {Gu, Huiya and Jinkerson, Robert E. and Davies, Fiona K. and Sisson, Lyle A. and Schneider, Philip E. and Posewitz, Matthew C.},
abstractNote = {The isolation or engineering of algal cells synthesizing high levels of medium-chain fatty acids (MCFAs) is attractive to mitigate the high clouding point of longer chain fatty acids in algal based biodiesel. To develop a more informed understanding of MCFA synthesis in photosynthetic microorganisms, we isolated several algae from Great Salt Lake and screened this collection for MCFA accumulation to identify strains naturally accumulating high levels of MCFA. A diatom, Chaetoceros sp. GSL56, accumulated particularly high levels of C14 (up to 40%), with the majority of C14 fatty acids allocated in triacylglycerols. Using whole cell transcriptome sequencing and de novo assembly, putative genes encoding fatty acid synthesis enzymes were identified. Enzymes from this Chaetoceros sp. were expressed in the cyanobacterium Synechococcus sp. PCC 7002 to validate gene function and to determine whether eukaryotic enzymes putatively lacking bacterial evolutionary control mechanisms could be used to improve MCFA production in this promising production strain. Replacement of the Synechococcus 7002 native FabH with a Chaetoceros ketoacyl-ACP synthase Ill increased MCFA synthesis up to fivefold. In conclusion, the level of increase is dependent on promoter strength and culturing conditions.},
doi = {10.3389/fpls.2016.00690},
journal = {Frontiers in Plant Science},
number = ,
volume = 7,
place = {United States},
year = {Thu May 26 00:00:00 EDT 2016},
month = {Thu May 26 00:00:00 EDT 2016}
}
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Works referenced in this record:
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Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes
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The potentials and challenges of algae based biofuels: A review of the techno-economic, life cycle, and resource assessment modeling
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Perspectives on engineering strategies for improving biofuel production from microalgae — A critical review
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Fatty acid and lipid class composition of the eicosapentaenoic acid-producing microalga, Nitzschia laevis
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Dependence of biodiesel fuel properties on the structure of fatty acid alkyl esters
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Marine metagenomics as a source for bioprospecting
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Transcriptome analysis of Chlamydomonas reinhardtii during the process of lipid accumulation
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Genetic engineering of fatty acid chain length in Phaeodactylum tricornutum
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Altered carbohydrate metabolism in glycogen synthase mutants of Synechococcus sp. strain PCC 7002: Cell factories for soluble sugars
journal, March 2013
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- Metabolic Engineering, Vol. 16
The initiation ketosynthase (FabH) is the sole rate-limiting enzyme of the fatty acid synthase of Synechococcus sp. PCC 7002
journal, March 2014
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- Metabolic Engineering, Vol. 22
“Designer” Biodiesel: Optimizing Fatty Ester Composition to Improve Fuel Properties †
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- Energy & Fuels, Vol. 22, Issue 2
Draft genome sequence and genetic transformation of the oleaginous alga Nannochloropsis gaditana
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Condensing enzymes from Cuphea wrightii associated with medium chain fatty acid biosynthesis
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Knockout of the regulatory site of 3-ketoacyl-ACP synthase III enhances short- and medium-chain acyl-ACP synthesis
journal, October 2000
- Abbadi, Amine; Brummel, Monika; Spener, Friedrich
- The Plant Journal, Vol. 24, Issue 1
Fatty acid production in genetically modified cyanobacteria
journal, April 2011
- Liu, X.; Sheng, J.; Curtiss III, R.
- Proceedings of the National Academy of Sciences, Vol. 108, Issue 17, p. 6899-6904
SOAP: short oligonucleotide alignment program
journal, January 2008
- Li, R.; Li, Y.; Kristiansen, K.
- Bioinformatics, Vol. 24, Issue 5
Phylogeny.fr: robust phylogenetic analysis for the non-specialist
journal, May 2008
- Dereeper, A.; Guignon, V.; Blanc, G.
- Nucleic Acids Research, Vol. 36, Issue Web Server
ABySS: A parallel assembler for short read sequence data
journal, February 2009
- Simpson, J. T.; Wong, K.; Jackman, S. D.
- Genome Research, Vol. 19, Issue 6
Using the miraEST Assembler for Reliable and Automated mRNA Transcript Assembly and SNP Detection in Sequenced ESTs
journal, May 2004
- Chevreux, Bastien; Pfisterer, Thomas; Drescher, Bernd
- Genome Research, Vol. 14, Issue 6, p. 1147-1159
Overexpression of 3-Ketoacyl-Acyl-Carrier Protein Synthase IIIs in Plants Reduces the Rate of Lipid Synthesis
journal, February 2001
- Dehesh, Katayoon; Tai, Heeyoung; Edwards, Patricia
- Plant Physiology, Vol. 125, Issue 2
A Comparison of the Metabolic Fate of Fatty Acids of Different Chain Lengths in Developing Oilseeds
journal, July 1989
- Battey, James F.; Ohlrogge, John B.
- Plant Physiology, Vol. 90, Issue 3
Fatty acid biosynthesis redirected to medium chains in transgenic oilseed plants
journal, July 1992
- Voelker, T.; Worrell, A.; Anderson, L.
- Science, Vol. 257, Issue 5066
Alteration of the specificity and regulation of fatty acid synthesis of Escherichia coli by expression of a plant medium-chain acyl-acyl carrier protein thioesterase.
journal, December 1994
- Voelker, T. A.; Davies, H. M.
- Journal of Bacteriology, Vol. 176, Issue 23, p. 7320-7327
Regulation of fatty acid biosynthesis in Escherichia coli.
journal, January 1993
- Magnuson, K.; Jackowski, S.; Rock, C. O.
- Microbiological Reviews, Vol. 57, Issue 3
Phylogenetic and experimental characterization of an acyl-ACP thioesterase family reveals significant diversity in enzymatic specificity and activity
journal, January 2011
- Jing, Fuyuan; Cantu, David C.; Tvaruzkova, Jarmila
- BMC Biochemistry, Vol. 12, Issue 1
The Marine Microbial Eukaryote Transcriptome Sequencing Project (MMETSP): Illuminating the Functional Diversity of Eukaryotic Life in the Oceans through Transcriptome Sequencing
journal, June 2014
- Keeling, Patrick J.; Burki, Fabien; Wilcox, Heather M.
- PLoS Biology, Vol. 12, Issue 6
Manipulating Fatty Acid Biosynthesis in Microalgae for Biofuel through Protein-Protein Interactions
journal, September 2012
- Blatti, Jillian L.; Beld, Joris; Behnke, Craig A.
- PLoS ONE, Vol. 7, Issue 9
Palmitoyl-Acyl Carrier Protein (ACP) Thioesterase and the Evolutionary Origin of Plant Acyl-ACP Thioesterases
journal, March 1995
- Jones, Aubrey; Davies, H. Maelor; Voelker, Toni A.
- The Plant Cell, Vol. 7, Issue 3
Improved Free Fatty Acid Production in Cyanobacteria with Synechococcus sp. PCC 7002 as Host
journal, May 2014
- Ruffing, Anne M.
- Frontiers in Bioengineering and Biotechnology, Vol. 2
Engineering Limonene and Bisabolene Production in Wild Type and a Glycogen-Deficient Mutant of Synechococcus sp. PCC 7002
journal, June 2014
- Davies, Fiona K.; Work, Victoria H.; Beliaev, Alexander S.
- Frontiers in Bioengineering and Biotechnology, Vol. 2
Lauric Acid Production in a Glycogen-Less Strain of Synechococcus sp. PCC 7002
journal, April 2015
- Work, Victoria H.; Melnicki, Matthew R.; Hill, Eric A.
- Frontiers in Bioengineering and Biotechnology, Vol. 3
Modeling sequencing errors by combining Hidden Markov models
text, January 2003
- Lottaz, Claudio; Iseli, Christian; Jongeneel, C. Victor
- Universität Regensburg
Works referencing / citing this record:
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