Modulation of medium-chain fatty acid synthesis in Synechococcus sp. PCC 7002 by replacing FabH with a Chaetoceros Ketoacyl-ACP synthase

Gu, Huiya; Jinkerson, Robert E.; Davies, Fiona K.; Sisson, Lyle A.; Schneider, Philip E.; Posewitz, Matthew C.

doi:10.3389/fpls.2016.00690

Title: Modulation of medium-chain fatty acid synthesis in Synechococcus sp. PCC 7002 by replacing FabH with a Chaetoceros Ketoacyl-ACP synthase

Journal Article · Thu May 26 00:00:00 EDT 2016 · Frontiers in Plant Science

DOI:https://doi.org/10.3389/fpls.2016.00690· OSTI ID:1281962

Gu, Huiya ^[1]; Jinkerson, Robert E. ^[2]; Davies, Fiona K. ^[1]; Sisson, Lyle A. ^[1]; Schneider, Philip E. ^[1]; Posewitz, Matthew C. ^[1]

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

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.

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Research Organization:: Colorado School of Mines, Golden, CO, (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22), Chemical Sciences, Geosciences and Biosciences Division

Grant/Contract Number:: SC0008547

OSTI ID:: 1281962

Journal Information:: Frontiers in Plant Science, Vol. 7; ISSN 1664-462X

Publisher:: Frontiers Research FoundationCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 8 works

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References (65)

Fatty acid production in genetically modified cyanobacteria Liu, X.; Sheng, J.; Curtiss III, R. Proceedings of the National Academy of Sciences, Vol. 108, Issue 17, p. 6899-6904 https://doi.org/10.1073/pnas.1103014108	journal	April 2011
Biocommodities from photosynthetic microorganisms Work, Victoria H.; Bentley, Fiona K.; Scholz, Matthew J. Environmental Progress & Sustainable Energy, Vol. 32, Issue 4 https://doi.org/10.1002/ep.11849	journal	September 2013
Genetic engineering of fatty acid chain length in Phaeodactylum tricornutum Radakovits, Randor; Eduafo, Patrick M.; Posewitz, Matthew C. Metabolic Engineering, Vol. 13, Issue 1 https://doi.org/10.1016/j.ymben.2010.10.003	journal	January 2011
Marine metagenomics as a source for bioprospecting Kodzius, Rimantas; Gojobori, Takashi Marine Genomics, Vol. 24 https://doi.org/10.1016/j.margen.2015.07.001	journal	December 2015
Selection, breeding and engineering of microalgae for bioenergy and biofuel production Larkum, Anthony W. D.; Ross, Ian L.; Kruse, Olaf Trends in Biotechnology, Vol. 30, Issue 4, p. 198-205 https://doi.org/10.1016/j.tibtech.2011.11.003	journal	April 2012
Manipulating Fatty Acid Biosynthesis in Microalgae for Biofuel through Protein-Protein Interactions Blatti, Jillian L.; Beld, Joris; Behnke, Craig A. PLoS ONE, Vol. 7, Issue 9 https://doi.org/10.1371/journal.pone.0042949	journal	September 2012
Contrasting Patterns of Community Assembly in the Stratified Water Column of Great Salt Lake, Utah Meuser, Jonathan E.; Baxter, Bonnie K.; Spear, John R. Microbial Ecology, Vol. 66, Issue 2 https://doi.org/10.1007/s00248-013-0180-9	journal	January 2013
Lauric Acid Production in a Glycogen-Less Strain of Synechococcus sp. PCC 7002 Work, Victoria H.; Melnicki, Matthew R.; Hill, Eric A. Frontiers in Bioengineering and Biotechnology, Vol. 3 https://doi.org/10.3389/fbioe.2015.00048	journal	April 2015
Fatty acid and lipid class composition of the eicosapentaenoic acid-producing microalga, Nitzschia laevis Chen, G.; Jiang, Y.; Chen, F. Food Chemistry, Vol. 104, Issue 4 https://doi.org/10.1016/j.foodchem.2007.03.008	journal	January 2007
A Comparison of the Metabolic Fate of Fatty Acids of Different Chain Lengths in Developing Oilseeds Battey, James F.; Ohlrogge, John B. Plant Physiology, Vol. 90, Issue 3 https://doi.org/10.1104/pp.90.3.835	journal	July 1989
Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes Kovach, Michael E.; Elzer, Philip H.; Steven Hill, D. Gene, Vol. 166, Issue 1 https://doi.org/10.1016/0378-1119(95)00584-1	journal	December 1995
The lipid composition of the unicellular green alga Chlamydomonas reinhardtii and the diatom Cyclotella meneghiniana investigated by MALDI-TOF MS and TLC Vieler, Astrid; Wilhelm, Christian; Goss, Reimund Chemistry and Physics of Lipids, Vol. 150, Issue 2 https://doi.org/10.1016/j.chemphyslip.2007.06.224	journal	December 2007
Effect of UV stress on the fatty acid and lipid class composition in two marine microalgae Pavlova lutheri (Pavlovophyceae) and Odontella aurita (Bacillariophyceae) Guihéneuf, Freddy; Fouqueray, Manuela; Mimouni, Virginie Journal of Applied Phycology, Vol. 22, Issue 5 https://doi.org/10.1007/s10811-010-9503-0	journal	February 2010
Fast and accurate short read alignment with Burrows-Wheeler transform Li, H.; Durbin, R. Bioinformatics, Vol. 25, Issue 14 https://doi.org/10.1093/bioinformatics/btp324	journal	May 2009
Modeling sequencing errors by combining Hidden Markov models Lottaz, C.; Iseli, C.; Jongeneel, C. V. Bioinformatics, Vol. 19, Issue Suppl 2 https://doi.org/10.1093/bioinformatics/btg1067	journal	September 2003
SOAP: short oligonucleotide alignment program Li, R.; Li, Y.; Kristiansen, K. Bioinformatics, Vol. 24, Issue 5 https://doi.org/10.1093/bioinformatics/btn025	journal	January 2008
Palmitoyl-acyl carrier protein (ACP) thioesterase and the evolutionary origin of plant acyl-ACP thioesterases. Jones, A.; Davies, H. M.; Voelker, T. A. The Plant Cell, Vol. 7, Issue 3 https://doi.org/10.1105/tpc.7.3.359	journal	March 1995
Altered carbohydrate metabolism in glycogen synthase mutants of Synechococcus sp. strain PCC 7002: Cell factories for soluble sugars Xu, Yu; Tiago Guerra, L.; Li, Zhongkui Metabolic Engineering, Vol. 16 https://doi.org/10.1016/j.ymben.2012.12.002	journal	March 2013
Using the miraEST Assembler for Reliable and Automated mRNA Transcript Assembly and SNP Detection in Sequenced ESTs Chevreux, Bastien; Pfisterer, Thomas; Drescher, Bernd Genome Research, Vol. 14, Issue 6, p. 1147-1159 https://doi.org/10.1101/gr.1917404	journal	May 2004
Engineering Limonene and Bisabolene Production in Wild Type and a Glycogen-Deficient Mutant of Synechococcus sp. PCC 7002 Davies, Fiona K.; Work, Victoria H.; Beliaev, Alexander S. Frontiers in Bioengineering and Biotechnology, Vol. 2 https://doi.org/10.3389/fbioe.2014.00021	journal	June 2014
“Designer” Biodiesel: Optimizing Fatty Ester Composition to Improve Fuel Properties ^† Knothe, Gerhard Energy & Fuels, Vol. 22, Issue 2 https://doi.org/10.1021/ef700639e	journal	March 2008
The Transcriptome and Proteome of the Diatom Thalassiosira pseudonana Reveal a Diverse Phosphorus Stress Response Dyhrman, Sonya T.; Jenkins, Bethany D.; Rynearson, Tatiana A. PLoS ONE, Vol. 7, Issue 3 https://doi.org/10.1371/journal.pone.0033768	journal	March 2012
Draft genome sequence and genetic transformation of the oleaginous alga Nannochloropsis gaditana Radakovits, Randor; Jinkerson, Robert E.; Fuerstenberg, Susan I. Nature Communications, Vol. 3, Issue 1 https://doi.org/10.1038/ncomms1688	journal	January 2012
The Marine Microbial Eukaryote Transcriptome Sequencing Project (MMETSP): Illuminating the Functional Diversity of Eukaryotic Life in the Oceans through Transcriptome Sequencing Keeling, Patrick J.; Burki, Fabien; Wilcox, Heather M. PLoS Biology, Vol. 12, Issue 6 https://doi.org/10.1371/journal.pbio.1001889	journal	June 2014
Transcriptome analysis of Chlamydomonas reinhardtii during the process of lipid accumulation Lv, Hexin; Qu, Ge; Qi, Xizhen Genomics, Vol. 101, Issue 4 https://doi.org/10.1016/j.ygeno.2013.01.004	journal	April 2013
Phylogeny.fr: robust phylogenetic analysis for the non-specialist Dereeper, A.; Guignon, V.; Blanc, G. Nucleic Acids Research, Vol. 36, Issue Web Server https://doi.org/10.1093/nar/gkn180	journal	May 2008
Phylogenetic and experimental characterization of an acyl-ACP thioesterase family reveals significant diversity in enzymatic specificity and activity Jing, Fuyuan; Cantu, David C.; Tvaruzkova, Jarmila BMC Biochemistry, Vol. 12, Issue 1 https://doi.org/10.1186/1471-2091-12-44	journal	January 2011
Changes in Transcript Abundance in Chlamydomonas reinhardtii following Nitrogen Deprivation Predict Diversion of Metabolism Miller, Rachel; Wu, Guangxi; Deshpande, Rahul R. Plant Physiology, Vol. 154, Issue 4 https://doi.org/10.1104/pp.110.165159	journal	October 2010
Evolution of acyl-ACP thioesterases and β-ketoacyl-ACP synthases revealed by protein–protein interactions Beld, Joris; Blatti, Jillian L.; Behnke, Craig Journal of Applied Phycology, Vol. 26, Issue 4 https://doi.org/10.1007/s10811-013-0203-4	journal	November 2013
Enzymatic assembly of DNA molecules up to several hundred kilobases Gibson, Daniel G.; Young, Lei; Chuang, Ray-Yuan Nature Methods, Vol. 6, Issue 5, p. 343-345 https://doi.org/10.1038/nmeth.1318	journal	April 2009
Improved Free Fatty Acid Production in Cyanobacteria with Synechococcus sp. PCC 7002 as Host Ruffing, Anne M. Frontiers in Bioengineering and Biotechnology, Vol. 2 https://doi.org/10.3389/fbioe.2014.00017	journal	May 2014
Fatty acid biosynthesis redirected to medium chains in transgenic oilseed plants Voelker, T.; Worrell, A.; Anderson, L. Science, Vol. 257, Issue 5066 https://doi.org/10.1126/science.1621095	journal	July 1992
The impact of nitrogen starvation on the dynamics of triacylglycerol accumulation in nine microalgae strains Breuer, Guido; Lamers, Packo P.; Martens, Dirk E. Bioresource Technology, Vol. 124 https://doi.org/10.1016/j.biortech.2012.08.003	journal	November 2012
Fatty acid composition of 12 microalgae for possible use in aquaculture feed Patil, Vishwanath; Källqvist, Torsten; Olsen, Elisabeth Aquaculture International, Vol. 15, Issue 1 https://doi.org/10.1007/s10499-006-9060-3	journal	July 2006
Fatty Acid Activation in Cyanobacteria Mediated by Acyl-Acyl Carrier Protein Synthetase Enables Fatty Acid Recycling Kaczmarzyk, Danuta; Fulda, Martin Plant Physiology, Vol. 152, Issue 3 https://doi.org/10.1104/pp.109.148007	journal	January 2010
Microalgae for biodiesel production and other applications: A review Mata, Teresa M.; Martins, António A.; Caetano, Nidia. S. Renewable and Sustainable Energy Reviews, Vol. 14, Issue 1, p. 217-232 https://doi.org/10.1016/j.rser.2009.07.020	journal	January 2010
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. Voelker, T. A.; Davies, H. M. Journal of Bacteriology, Vol. 176, Issue 23, p. 7320-7327 https://doi.org/10.1128/jb.176.23.7320-7327.1994	journal	December 1994
Dependence of biodiesel fuel properties on the structure of fatty acid alkyl esters Knothe, Gerhard Fuel Processing Technology, Vol. 86, Issue 10 https://doi.org/10.1016/j.fuproc.2004.11.002	journal	June 2005
Knockout of the regulatory site of 3-ketoacyl-ACP synthase III enhances short- and medium-chain acyl-ACP synthesis Abbadi, Amine; Brummel, Monika; Spener, Friedrich The Plant Journal, Vol. 24, Issue 1 https://doi.org/10.1046/j.1365-313x.2000.00841.x	journal	October 2000
Second Generation Biofuels: High-Efficiency Microalgae for Biodiesel Production Schenk, Peer M.; Thomas-Hall, Skye R.; Stephens, Evan BioEnergy Research, Vol. 1, Issue 1 https://doi.org/10.1007/s12155-008-9008-8	journal	March 2008
Perspectives on engineering strategies for improving biofuel production from microalgae — A critical review Ho, Shih-Hsin; Ye, Xiaoting; Hasunuma, Tomohisa Biotechnology Advances, Vol. 32, Issue 8 https://doi.org/10.1016/j.biotechadv.2014.09.002	journal	December 2014
The initiation ketosynthase (FabH) is the sole rate-limiting enzyme of the fatty acid synthase of Synechococcus sp. PCC 7002 Kuo, James; Khosla, Chaitan Metabolic Engineering, Vol. 22 https://doi.org/10.1016/j.ymben.2013.12.008	journal	March 2014
Metabolic Engineering of Fatty Acid Biosynthesis in Plants Thelen, Jay J.; Ohlrogge, John B. Metabolic Engineering, Vol. 4, Issue 1 https://doi.org/10.1006/mben.2001.0204	journal	January 2002
Regulation of fatty acid biosynthesis in Escherichia coli. Magnuson, K.; Jackowski, S.; Rock, C. O. Microbiological Reviews, Vol. 57, Issue 3 https://doi.org/10.1128/MMBR.57.3.522-542.1993	journal	January 1993
ABySS: A parallel assembler for short read sequence data Simpson, J. T.; Wong, K.; Jackman, S. D. Genome Research, Vol. 19, Issue 6 https://doi.org/10.1101/gr.089532.108	journal	February 2009
Novel kingdom-level eukaryotic diversity in anoxic environments Dawson, S. C.; Pace, N. R. Proceedings of the National Academy of Sciences, Vol. 99, Issue 12 https://doi.org/10.1073/pnas.062169599	journal	June 2002
Overexpression of 3-Ketoacyl-Acyl-Carrier Protein Synthase IIIs in Plants Reduces the Rate of Lipid Synthesis Dehesh, Katayoon; Tai, Heeyoung; Edwards, Patricia Plant Physiology, Vol. 125, Issue 2 https://doi.org/10.1104/pp.125.2.1103	journal	February 2001
Condensing enzymes from Cuphea wrightii associated with medium chain fatty acid biosynthesis Slabaugh, Mary B.; Leonard, Jeffrey M.; Knapp, Steven J. The Plant Journal, Vol. 13, Issue 5 https://doi.org/10.1046/j.1365-313x.1998.00065.x	journal	March 1998
Blast2GO: A Comprehensive Suite for Functional Analysis in Plant Genomics Conesa, Ana; Götz, Stefan International Journal of Plant Genomics, Vol. 2008 https://doi.org/10.1155/2008/619832	journal	January 2008
The role of β-ketoacyl-acyl carrier protein synthase III in the condensation steps of fatty acid biosynthesis in sunflower González-Mellado, Damián; von Wettstein-Knowles, Penny; Garcés, Rafael Planta, Vol. 231, Issue 6 https://doi.org/10.1007/s00425-010-1131-z	journal	March 2010
The potentials and challenges of algae based biofuels: A review of the techno-economic, life cycle, and resource assessment modeling Quinn, Jason C.; Davis, Ryan Bioresource Technology, Vol. 184 https://doi.org/10.1016/j.biortech.2014.10.075	journal	May 2015
Genetic engineering for plant oils: potential and limitations Battey, J. Trends in Biotechnology, Vol. 7, Issue 5 https://doi.org/10.1016/0167-7799(89)90088-7	journal	May 1989
Acyl lipid composition variation related to culture age and nitrogen concentration in continuous culture of the microalga Phaeodactylum tricornutum Alonso, Diego López; Belarbi, El-Hassan; Fernández-Sevilla, José M. Phytochemistry, Vol. 54, Issue 5 https://doi.org/10.1016/S0031-9422(00)00084-4	journal	June 2000
Three Acyltransferases and Nitrogen-responsive Regulator Are Implicated in Nitrogen Starvation-induced Triacylglycerol Accumulation in Chlamydomonas Boyle, Nanette R.; Page, Mark Dudley; Liu, Bensheng Journal of Biological Chemistry, Vol. 287, Issue 19 https://doi.org/10.1074/jbc.M111.334052	journal	March 2012
Plant triacylglycerols as feedstocks for the production of biofuels Durrett, Timothy P.; Benning, Christoph; Ohlrogge, John The Plant Journal, Vol. 54, Issue 4, p. 593-607 https://doi.org/10.1111/j.1365-313X.2008.03442.x	journal	May 2008
A Cuphea β‐ketoacyl‐ACP synthase shifts the synthesis of fatty acids towards shorter chains in Arabidopsis seeds expressing Cuphea FatB thioesterases Leonard, Jeffrey M.; Knapp, Steven J.; Slabaugh, Mary B. The Plant Journal, Vol. 13, Issue 5 https://doi.org/10.1046/j.1365-313X.1998.00066.x	journal	March 1998
The gross chemical composition and fatty acid composition of 18 species of tropical Australian microalgae for possible use in mariculture Renaud, Susan M.; Thinh, Luong-Van; Parry, David L. Aquaculture, Vol. 170, Issue 2 https://doi.org/10.1016/S0044-8486(98)00399-8	journal	January 1999
Changes in the Lipid Composition of Thalassiosira pseudonana during Its Life Cycle Zhukova, N. V. Russian Journal of Plant Physiology, Vol. 51, Issue 5 https://doi.org/10.1023/B:RUPP.0000040759.04882.8c	journal	September 2004
Palmitoyl-Acyl Carrier Protein (ACP) Thioesterase and the Evolutionary Origin of Plant Acyl-ACP Thioesterases Jones, Aubrey; Davies, H. Maelor; Voelker, Toni A. The Plant Cell, Vol. 7, Issue 3 https://doi.org/10.2307/3869857	journal	March 1995
Modeling sequencing errors by combining Hidden Markov models Lottaz, Claudio; Iseli, Christian; Jongeneel, C. Victor Universität Regensburg https://doi.org/10.5283/epub.32949	text	January 2003
The Marine Microbial Eukaryote Transcriptome Sequencing Project (MMETSP): Illuminating the Functional Diversity of Eukaryotic Life in the Oceans through Transcriptome Sequencing Keeling, Patrick J.; Burki, Fabien; Wilcox, Heather M. Columbia University https://doi.org/10.7916/d8445mrv	text	January 2014
How can we genetically engineer oilseed crops to produce high levels of medium-chain fatty acids? Dehesh, Katayoon European Journal of Lipid Science and Technology, Vol. 103, Issue 10 https://doi.org/10.1002/1438-9312(200110)103:10<688::aid-ejlt688>3.0.co;2-9	journal	October 2001
Machine learning discovery of missing links that mediate alternative branches to plant alkaloids Vavricka, Christopher J.; Takahashi, Shunsuke; Watanabe, Naoki Nature Communications, Vol. 13, Issue 1 https://doi.org/10.1038/s41467-022-28883-8	journal	March 2022
Targeted A-to-G base editing of chloroplast DNA in plants Mok, Young Geun; Hong, Sunghyun; Bae, Su-Ji Nature Plants, Vol. 8, Issue 12 https://doi.org/10.1038/s41477-022-01279-8	journal	December 2022
Regulation of fatty acid biosynthesis in Escherichia coli. Magnuson, K.; Jackowski, S.; Rock, C. O. Microbiological Reviews, Vol. 57, Issue 3 https://doi.org/10.1128/mr.57.3.522-542.1993	journal	January 1993

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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
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Title: Modulation of medium-chain fatty acid synthesis in Synechococcus sp. PCC 7002 by replacing FabH with a Chaetoceros Ketoacyl-ACP synthase

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