Abstract
Interest in microalgal biotechnology for the production of bioactive and nutritional ingredients has increased tremendously with the urgent need for developing renewable bioresources. Microalgae represent a huge and still insufficiently tapped resource of LC-PUFA for human nutrition and health-related applications. Photosynthetic omega-3 and omega-6 LC-PUFA-producing microalgae are potent organisms and synthetic biology chassis for the production of high-value constituents for both health and aquaculture sectors. This chapter covers the diversity of eukaryotic microalgae in relation to their LC-PUFA production and outlines the fundamental role of microalgae as primary producers of LC-PUFA. We provide mechanistic insights into the biosynthesis of LC-PUFA in photosynthetic microalgae and highlight their biotechnological applications. The ongoing research aims to understand the molecular mechanisms by which microalgae of different evolutionary groups synthesize, relocate and incorporate LC-PUFA into complex lipids in the context of their multifaceted cellular organization. This research is supported by the availability of genomic information and ‘omics’ studies in a growing number of species. Recent progress in microalgal genetic transformation, genome editing and metabolic engineering has enabled the manipulation of LC-PUFA biosynthesis. We discuss the prospects for LC-PUFA manufacturing by microalgal biotechnology as a renewable and sustainable alternative to the finite resources of LC-PUFA.
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Aarsetoey H, Aarsetoey R, Lindner T et al (2011) Low levels of the omega-3 index are associated with sudden cardiac arrest and remain stable in survivors in the subacute phase. Lipids 46:151–161. https://doi.org/10.1007/s11745-010-3511-3
Abida H, Dolch L-J, Meï C et al (2015) Membrane glycerolipid remodeling triggered by nitrogen and phosphorus starvation in Phaeodactylum tricornutum. Plant Physiol 167:118–136. https://doi.org/10.1104/pp.114.252395
Agostoni C (2008) Role of long-chain polyunsaturated fatty acids in the first year of life. J Pediatr Gastroenterol Nutr 47:S41–S44. https://doi.org/10.1097/01.mpg.0000338811.52062.b2
Ajjawi I, Verruto J, Aqui M et al (2017) Lipid production in Nannochloropsis gaditana is doubled by decreasing expression of a single transcriptional regulator. Nat Biotechnol 35:647–652. https://doi.org/10.1038/nbt.3865
Alves SP, Mendonça SH, Silva JL, Bessa RJB (2018) Nannochloropsis oceanica, a novel natural source of rumen-protected eicosapentaenoic acid (EPA) for ruminants. Sci Rep 8:10269. https://doi.org/10.1038/s41598-018-28576-7
Andrianasolo EH, Haramaty L, Vardi A et al (2008) Apoptosis-inducing galactolipids from a cultured marine diatom, Phaeodactylum tricornutum. J Nat Prod 71:1197–1201. https://doi.org/10.1021/np800124k.Apoptosis-Inducing
Apt KE, Grossman AR, Kroth-Pancic PG (1996) Stable nuclear transformation of the diatom Phaeodactylum tricornutum. Mol Gen Genet 252:572–579. https://doi.org/10.1007/BF02172403
Arao T, Yamada M (1994) Biosynthesis of polyunsaturated fatty acids in the marine diatom, Phaeodactylum tricornutum. Phytochemistry 35:1177–1181. https://doi.org/10.1016/S0031-9422(00)94817-9
Aussant J, Guihéneuf F, Stengel DB (2018) Impact of temperature on fatty acid composition and nutritional value in eight species of microalgae. Appl Microbiol Biotechnol 102:5279–5297. https://doi.org/10.1007/s00253-018-9001-x
Balamurugan S, Wang X, Wang H-L et al (2017) Occurrence of plastidial triacylglycerol synthesis and the potential regulatory role of AGPAT in the model diatom Phaeodactylum tricornutum. Biotechnol Biofuels 10:97. https://doi.org/10.1186/s13068-017-0786-0
Barclay WR, Meager KM, Abril JR (1994) Heterotrophic production of long chain omega-3 fatty acids utilizing algae and algae-like microorganisms. J Appl Phycol 6:123–129. https://doi.org/10.1007/BF02186066
Beacham TA, Macia VM, Rooks P et al (2015) Altered lipid accumulation in Nannochloropsis salina CCAP849/3 following EMS and UV induced mutagenesis. Biotechnol Rep (Amst) 7:87–94. https://doi.org/10.1016/j.btre.2015.05.007
Bell MV, Pond D (1996) Lipid composition during growth of motile and coccolith forms of Emiliania huxleyi. Phytochemistry 41:465–471. https://doi.org/10.1016/0031-9422(95)00663-X
Bell JG, Sargent JR (2003) Arachidonic acid in aquaculture feeds: current status and future opportunities. Aquaculture 218:491–499. https://doi.org/10.1016/S0044-8486(02)00370-8
Bendif EM, Probert I, Schroeder DC, de Vargas C (2013) On the description of Tisochrysis lutea gen. nov. sp. nov. and Isochrysis nuda sp. nov. in the Isochrysidales, and the transfer of Dicrateria to the Prymnesiales (Haptophyta). J Appl Phycol 25:1763–1776. https://doi.org/10.1007/s10811-013-0037-0
Betancor MB, Sprague M, Sayanova O et al (2015) Evaluation of a high-EPA oil from transgenic Camelina sativa in feeds for Atlantic salmon (Salmo salar L.): effects on tissue fatty acid composition, histology and gene expression. Aquaculture 444:1–12. https://doi.org/10.1016/j.aquaculture.2015.03.020
Betancor MB, Sprague M, Montero D et al (2016) Replacement of marine fish oil with de novo omega-3 oils from transgenic Camelina sativa in feeds for gilthead sea bream (Sparus aurata L.). Lipids 51:1171–1191. https://doi.org/10.1007/s11745-016-4191-4
Bhattacharya D, Price DC, Chan CX et al (2013) Genome of the red alga Porphyridium purpureum. Nat Commun 4:1941. https://doi.org/10.1038/ncomms2931
Bigogno C, Khozin-Goldberg I, Adlerstein D, Cohen Z (2002a) Biosynthesis of arachidonic acid in the oleaginous microalga Parietochloris incisa (Chlorophyceae): radiolabeling studies. Lipids 37:209–216. https://doi.org/10.1007/s11745-002-0882-6
Bigogno C, Khozin-Goldberg I, Boussiba S et al (2002b) Lipid and fatty acid composition of the green oleaginous alga Parietochloris incisa, the richest plant source of arachidonic acid. Phytochemistry 60:497–503. https://doi.org/10.1016/S0031-9422(02)00100-0
Bigogno C, Khozin-Goldberg I, Cohen Z (2002c) Accumulation of arachidonic acid-rich triacylglycerols in the microalga Parietochloris incisa (Trebouxiophyceae, Chlorophyta). Phytochemistry 60:135–143. https://doi.org/10.1016/S0031-9422(02)00037-7
Boller T, Felix G (2009) A renaissance of elicitors: perception of microbe-associated molecular patterns and danger signals by pattern-recognition receptors. Annu Rev Plant Biol 60:379–406. https://doi.org/10.1146/annurev.arplant.57.032905.105346
Boussiba S, Vonshak A, Cohen Z et al (1987) Lipid and biomass production by the halotolerant microalga Nannochloropsis salina. Biomass 12:37–47. https://doi.org/10.1016/0144-4565(87)90006-0
Brodie J, Chan CX, De Clerck O et al (2017) The algal revolution. Trends Plant Sci 22:726–738
Burja AM, Radianingtyas H, Windust A, Barrow CJ (2006) Isolation and characterization of polyunsaturated fatty acid producing Thraustochytrium species: screening of strains and optimization of omega-3 production. Appl Microbiol Biotechnol 72:1161–1169. https://doi.org/10.1007/s00253-006-0419-1
Calder PC (2015) Marine omega-3 fatty acids and inflammatory processes: effects, mechanisms and clinical relevance. BBA – Mol Cell Biol Lipids 1851:469–484. https://doi.org/10.1016/j.bbalip.2014.08.010
Camacho-Rodríguez J, Cerón-García MC, Fernández-Sevilla JM, Molina-Grima E (2015) Genetic algorithm for the medium optimization of the microalga Nannochloropsis gaditana cultured to aquaculture. Bioresour Technol 177:102–109. https://doi.org/10.1016/j.biortech.2014.11.057
Carrier G, Garnier M, Le Cunff L et al (2014) Comparative transcriptome of wild type and selected strains of the microalgae Tisochrysis lutea provides insights into the genetic basis, lipid metabolism and the life cycle. PLoS One 9:e86889. https://doi.org/10.1371/journal.pone.0086889
Chaturvedi R, Fujita Y (2006) Isolation of enhanced eicosapentaenoic acid producing mutants of Nannochloropsis oculata ST-6 using ethyl methane sulfonate induced mutagenesis techniques and their characterization at mRNA transcript level. Phycol Res 54:208–219. https://doi.org/10.1111/j.1440-1835.2006.00428.x
Chaturvedi R, Uppalapati SR, Alamsjah MA, Fujita Y (2004) Isolation of quizalofop-resistant mutants of Nannochloropsis oculata (Eustigmatophyceae) with high eicosapentaenoic acid following N-methyl-N-nitrosourea-induced random mutagenesis. J Appl Phycol 16:135–144. https://doi.org/10.1023/B:JAPH.0000044826.70360.8e
Chauton MS, Reitan KI, Norsker NH et al (2015) A techno-economic analysis of industrial production of marine microalgae as a source of EPA and DHA-rich raw material for aquafeed: research challenges and possibilities. Aquaculture 436:95–103. https://doi.org/10.1016/j.aquaculture.2014.10.038
Cheng J, Feng J, Cheng R et al (2014) Gene expression and metabolic pathways related to cell growth and lipid synthesis in diatom Nitzschia ZJU2 after two rounds of mutagenesis by γ-rays. RSC Adv 4:28463–28470. https://doi.org/10.1039/C4RA03359K
Conte M, Lupette J, Seddiki K et al (2018) Screening for biologically annotated drugs that trigger triacylglycerol accumulation in the diatom phaeodactylum. Plant Physiol 177:532–552. https://doi.org/10.1104/pp.17.01804
Cook O, Hildebrand M (2016) Enhancing LC-PUFA production in Thalassiosira pseudonana by overexpressing the endogenous fatty acid elongase genes. J Appl Phycol 28:897–905. https://doi.org/10.1007/s10811-015-0617-2
Crozet P, Navarro FJ, Willmund F et al (2018) Birth of a photosynthetic chassis: a MoClo toolkit enabling synthetic biology in the microalga Chlamydomonas reinhardtii. ACS Synth Biol 7:2074–2086. https://doi.org/10.1021/acssynbio.8b00251
Daboussi F, Leduc S, Maréchal A et al (2014) Genome engineering empowers the diatom Phaeodactylum tricornutum for biotechnology. Nat Commun 5:3831. https://doi.org/10.1038/ncomms4831
Dagar A, Zilberg D, Cohen Z et al (2010) Short-term dietary supplementation with the microalga Parietochloris incisa enhances stress resistance in guppies Poecilia reticulata. Aquac Res 41:267–277. https://doi.org/10.1111/j.1365-2109.2009.02329.x
Das UN (2006) Essential fatty acids: biochemistry, physiology and pathology. Biotechnol J 1:420–439. https://doi.org/10.1002/biot.200600012
Desbois AP, Mearns-Spragg A, Smith VJ (2009) A fatty acid from the diatom Phaeodactylum tricornutum is antibacterial against diverse bacteria including multi-resistant Staphylococcus aureus (MRSA). Mar Biotechnol 11:45–52. https://doi.org/10.1007/s10126-008-9118-5
Diner RE, Bielinski VA, Dupont CL et al (2016) Refinement of the diatom episome maintenance sequence and improvement of conjugation-based DNA delivery methods. Front Bioeng Biotechnol 4:65. https://doi.org/10.3389/fbioe.2016.00065
Dolch L-J, Maréchal E (2015) Inventory of fatty acid desaturases in the pennate diatom Phaeodactylum tricornutum. Mar Drugs 13:1317–1339. https://doi.org/10.3390/md13031317
Dolch L-J, Rak C, Perin G et al (2017) A palmitic acid elongase affects eicosapentaenoic acid and plastidial monogalactosyldiacylglycerol levels in Nannochloropsis. Plant Physiol 173:742–759. https://doi.org/10.1104/pp.16.01420
Domergue F, Lerchl J, Zähringer U, Heinz E (2002) Cloning and functional characterization of Phaeodactylum tricornutum front-end desaturases involved in eicosapentaenoic acid biosynthesis. Eur J Biochem 269:4105–4113
Domergue F, Abbadi A, Ott C et al (2003) Acyl carriers used as substrates by the desaturases and elongases involved in very long-chain polyunsaturated fatty acids biosynthesis reconstituted in yeast. J Biol Chem 278:35115–35126. https://doi.org/10.1074/jbc.M305990200
Domergue F, Abbadi A, Zähringer U et al (2005) In vivo characterization of the first acyl-CoA Delta6-desaturase from a member of the plant kingdom, the microalga Ostreococcus tauri. Biochem J 389:483–490. https://doi.org/10.1042/BJ20050111
Draaisma RB, Wijffels RH, (Ellen) Slegers P et al (2013) Food commodities from microalgae. Curr Opin Biotechnol 24:169–177. https://doi.org/10.1016/J.COPBIO.2012.09.012
Dragosits M, Mattanovich D (2013) Adaptive laboratory evolution – principles and applications for biotechnology. Microb Cell Fact 12:64. https://doi.org/10.1186/1475-2859-12-64
Dunahay TG, Jarvis EE, Roessler PG (1995) Genetic transformation of the diatoms Cyclotella cryptica and Navicula saprophila. J Phycol 31:1004–1012. https://doi.org/10.1111/j.0022-3646.1995.01004.x
Dunahay TG, Jarvis EE, Dais SS et al (1996) Manipulation of microalgal lipid production using genetic engineering. Appl Biochem Biotechnol 57:223. https://doi.org/10.1007/BF02941703
Endo H, Yoshida M, Uji T et al (2016) Stable nuclear transformation system for the coccolithophorid alga Pleurochrysis carterae. Sci Rep 6:1–10. https://doi.org/10.1038/srep22252
FAO (2016) The state of world fisheries and aquaculture 2016
Fischer H, Robl I, Sumper M, Kroger N (1999) Targeting and covalent modification of cell wall and membrane proteins heterologously expressed in the diatom Cylindrotheca fusiformis (Bacillariophyceae). J Phycol 35:113–120. https://doi.org/10.1046/j.1529-8817.1999.3510113.x
Freire I, Cortina-Burgueño A, Grille P et al (2016) Nannochloropsis limnetica: a freshwater microalga for marine aquaculture. Aquaculture 459:124–130. https://doi.org/10.1016/J.AQUACULTURE.2016.03.015
Frigolet ME, Gutiérrez-Aguilar R (2017) The role of the novel lipokine palmitoleic acid in health and disease. Adv Nutr An Int Rev J 8:173S–181S. https://doi.org/10.3945/an.115.011130
Fu HY, Picot D, Choquet Y et al (2017) Redesigning the QA binding site of Photosystem II allows reduction of exogenous quinones. Nat Commun 8:15274. https://doi.org/10.1038/ncomms15274
Fukuchi-Mizutani M, Tasaka Y, Tanaka Y et al (1998) Characterization of delta 9 acyl-lipid desaturase homologues from Arabidopsis thaliana. Plant Cell Physiol 39:247–253
Funk CD (2001) Prostaglandins and leukotrienes: advances in eicosanoid biology. Science 294:1871–1875
García JL, de Vicente M, Galán B (2017) Microalgae, old sustainable food and fashion nutraceuticals. Microb Biotechnol 10:1017–1024. https://doi.org/10.1111/1751-7915.12800
Georgianna DR, Mayfield SP (2012) Exploiting diversity and synthetic biology for the production of algal biofuels. Nature 488:329–335. https://doi.org/10.1038/nature11479
Gerber M (2012) Omega-3 fatty acids and cancers: a systematic update review of epidemiological studies. Br J Nutr 107:S228–S239. https://doi.org/10.1017/S0007114512001614
Gill S, Willette S, Dungan B et al (2018) Suboptimal temperature acclimation affects Kennedy pathway gene expression, lipidome and metabolite profile of Nannochloropsis salina during PUFA enriched TAG synthesis. Mar Drugs 16:425. https://doi.org/10.3390/md16110425
Gong Y, Guo X, Wan X et al (2011) Characterization of a novel thioesterase (PtTE) from Phaeodactylum tricornutum. J Basic Microbiol 51:666–672. https://doi.org/10.1002/jobm.201000520
Guarnieri MT, Pienkos PT (2015) Algal omics: unlocking bioproduct diversity in algae cell factories. Photosynth Res 123:255–263
Guihéneuf F, Stengel D (2013) LC-PUFA-enriched oil production by microalgae: accumulation of lipid and triacylglycerols containing n-3 LC-PUFA is triggered by nitrogen limitation and inorganic carbon availability in the marine haptophyte Pavlova lutheri. Mar Drugs 11:4246–4266. https://doi.org/10.3390/md11114246
Guihéneuf F, Mimouni V, Ulmann L, Tremblin G (2009) Combined effects of irradiance level and carbon source on fatty acid and lipid class composition in the microalga Pavlova lutheri commonly used in mariculture. J Exp Mar Bio Ecol 369:136–143. https://doi.org/10.1016/J.JEMBE.2008.11.009
Guihéneuf F, Fouqueray M, Mimouni V et al (2010) Effect of UV stress on the fatty acid and lipid class composition in two marine microalgae Pavlova lutheri (Pavlovophyceae) and Odontella aurita (Bacillariophyceae). J Appl Phycol 22:629–638. https://doi.org/10.1007/s10811-010-9503-0
Guschina IA, Harwood JL (2006) Lipids and lipid metabolism in eukaryotic algae. Prog Lipid Res 45:160–186. https://doi.org/10.1016/j.plipres.2006.01.001
Haimeur A, Ulmann L, Mimouni V et al (2012) The role of Odontella aurita, a marine diatom rich in EPA, as a dietary supplement in dyslipidemia, platelet function and oxidative stress in high-fat fed rats. Lipids Health Dis 11:147. https://doi.org/10.1186/1476-511X-11-147
Hamilton ML, Haslam RP, Napier JA, Sayanova O (2014) Metabolic engineering of Phaeodactylum tricornutum for the enhanced accumulation of omega-3 long chain polyunsaturated fatty acids. Metab Eng 22:3–9. https://doi.org/10.1016/j.ymben.2013.12.003
Hamilton ML, Warwick J, Terry A et al (2015) Towards the industrial production of omega-3 long chain polyunsaturated fatty acids from a genetically modified diatom Phaeodactylum tricornutum. PLoS One 10:e0144054. https://doi.org/10.1371/journal.pone.0144054
Harwood JL, Jones AL (1989) Lipid metabolism in algae. In: Callow JA (ed) Advances in botanical research. Academic, London, pp 1–53
Haslam RP, Sayanova O, Kim HJ et al (2016) Synthetic redesign of plant lipid metabolism. Plant J 87:76–86. https://doi.org/10.1111/tpj.13172
Henley WJ, Litaker RW, Novoveská L et al (2013) Initial risk assessment of genetically modified (GM) microalgae for commodity-scale biofuel cultivation. Algal Res 2:66–77. https://doi.org/10.1016/J.ALGAL.2012.11.001
Hodgson PA, Henderson RJ, Sargent JR, Leftley JW (1991) Patterns of variation in the lipid class and fatty acid composition of Nannochloropsis oculata (Eustigmatophyceae) during batch culture. J Appl Phycol 3:169–181. https://doi.org/10.1007/BF00003699
Hoffmann M, Wagner M, Abbadi A et al (2008) Metabolic engineering of ω3-very long chain polyunsaturated fatty acid production by an exclusively acyl-CoA-dependent pathway. J Biol Chem 283:22352–22362. https://doi.org/10.1074/jbc.M802377200
Hopes A, Nekrasov V, Kamoun S, Mock T (2016) Editing of the urease gene by CRISPR-Cas in the diatom Thalassiosira pseudonana. Plant Methods 12. https://doi.org/10.1186/s13007-016-0148-0
Horrocks LA, Yeo YK (1999) Health benefits of docosahexaenoic acid (DHA). Pharmacol Res 40:211–225
Hsiao TY, Blanch HW (2005) Physiological studies of eicosapentaenoic acid production in the marine microalga Glossomastix chrysoplasta. Biotechnol Bioeng 93:465–475. https://doi.org/10.1002/bit.20761
Huang W, Daboussi F (2017) Genetic and metabolic engineering in diatoms. Philos Trans R Soc B Biol Sci 372:20160411. https://doi.org/10.1098/rstb.2016.0411
Hubert F, Poisson L, Loiseau C et al (2017) Lipids and lipolytic enzymes of the microalga Isochrysis galbana. OCL 24:D407. https://doi.org/10.1051/ocl/2017023
Huerlimann R, de Nys R, Heimann K (2010) Growth, lipid content, productivity, and fatty acid composition of tropical microalgae for scale-up production. Biotechnol Bioeng 107:245–257. https://doi.org/10.1002/bit.22809
Hulatt CJ, Wijffels RH, Bolla S, Kiron V (2017) Production of fatty acids and protein by nannochloropsis in flat-plate photobioreactors. PLoS One 12:e0170440. https://doi.org/10.1371/journal.pone.0170440
Ifuku K, Yan D, Miyahara M et al (2015) A stable and efficient nuclear transformation system for the diatom Chaetoceros gracilis. Photosynth Res 123:203–211. https://doi.org/10.1007/s11120-014-0048-y
Innis SM (2000) The role of dietary n–6 and n–3 fatty acids in the developing brain. Dev Neurosci 22:474–480. https://doi.org/10.1159/000017478
Iskandarov U, Khozin-Goldberg I, Ofir R, Cohen Z (2009) Cloning and characterization of the Δ6 polyunsaturated fatty acid elongase from the green microalga Parietochloris incisa. Lipids 44:545–554. https://doi.org/10.1007/s11745-009-3301-y
Iskandarov U, Khozin-Goldberg I, Cohen Z (2010) Identification and characterization of Δ12, Δ6, and Δ5 desaturases from the green microalga Parietochloris incisa. Lipids 45:519–530. https://doi.org/10.1007/s11745-010-3421-4
Iskandarov U, Khozin-Goldberg I, Cohen Z (2011) Selection of a DGLA-producing mutant of the microalga Parietochloris incisa: I. Identification of mutation site and expression of VLC-PUFA biosynthesis genes. Appl Microbiol Biotechnol 90:249–256. https://doi.org/10.1007/s00253-010-3005-5
Jamers A, Blust R, De Coen W (2009) Omics in algae: paving the way for a systems biological understanding of algal stress phenomena? Aquat Toxicol 92:114–121. https://doi.org/10.1016/j.aquatox.2009.02.012
Janssen JH, Lamers PP, de Vos RCH et al (2019) Translocation and de novo synthesis of eicosapentaenoic acid (EPA) during nitrogen starvation in Nannochloropsis gaditana. Algal Res 37:138–144. https://doi.org/10.1016/j.algal.2018.11.025
Jiang M, Guo B, Wan X et al (2014) Isolation and characterization of the diatom phaeodactylum Δ5-elongase gene for transgenic LC-PUFA production in Pichia pastoris. Mar Drugs 12:1317–1334. https://doi.org/10.3390/md12031317
Jinkerson RE, Radakovits R, Posewitz MC (2013) Genomic insights from the oleaginous model alga Nannochloropsis gaditana. Bioengineered 4:37–43. https://doi.org/10.4161/bioe.21880
Karas BJ, Diner RE, Lefebvre SC et al (2015) Designer diatom episomes delivered by bacterial conjugation. Nat Commun 6:6925. https://doi.org/10.1038/ncomms7925
Kaulmann U, Hertweck C (2002) Biosynthesis of polyunsaturated fatty acids by polyketide synthases. Angew Chem Int Ed 41:1866. https://doi.org/10.1002/1521-3773
Kaye Y, Grundman O, Leu S et al (2015) Metabolic engineering toward enhanced LC-PUFA biosynthesis in Nannochloropsis oceanica: overexpression of endogenous Δ12 desaturase driven by stress-inducible promoter leads to enhanced deposition of polyunsaturated fatty acids in TAG. Algal Res 11:387–398. https://doi.org/10.1016/j.algal.2015.05.003
Khozin I, Adlerstein D, Bigogno C et al (1997) Elucidation of the biosynthesis of eicosapentaenoic acid in the microalga Porphyridium cruentum. Plant Physiol 114:223–230
Khozin-Goldberg I, Yu HZ, Adlerstein D et al (2000) Triacylglycerols of the red microalga Porphyridium cruentum can contribute to the biosynthesis of eukaryotic galactolipids. Lipids 35:881–889
Khozin-Goldberg I, Bigogno C, Shrestha P, Cohen Z (2002a) Nitrogen starvation induces the accumulation of arachidonic acid in the freshwater green alga Parietochloris incisa (Trebouxiophyceae). J Phycol 38:1–5. https://doi.org/10.1046/j.1529-8817.2002.01160.x
Khozin-Goldberg I, Didi-Cohen S, Shayakhmetova I, Cohen Z (2002b) Biosynthesis of eicosapentaenoic acid (EPA) in the freshwater eustigmatophyte Monodus subterraneus (Eustigmatophyceae). J Phycol 38:745–756. https://doi.org/10.1046/j.1529-8817.2002.02006.x
Khozin-Goldberg I, Iskandarov U, Cohen Z (2011) LC-PUFA from photosynthetic microalgae: occurrence, biosynthesis, and prospects in biotechnology. Appl Microbiol Biotechnol 91:905–915
Khozin-Goldberg I, Leu S, Boussiba S (2016) Microalgae as a source for VLC-PUFA production. In: Nakamura Y, Li-Beisson Y (eds) Lipids in plant and algae development, subcellular biochemistry. Springer, Cham, pp 471–510
Kilian O, Vick B (2013) United States Patent US 8.440,805 B2
Kilian O, Benemann CSE, Niyogi KK, Vick B (2011) High-efficiency homologous recombination in the oil-producing alga Nannochloropsis sp. Proc Natl Acad Sci 108:21265–21269. https://doi.org/10.1073/pnas.1105861108
Kiron V, Phromkunthong W, Huntley M et al (2012) Marine microalgae from biorefinery as a potential feed protein source for Atlantic salmon, common carp and whiteleg shrimp. Aquac Nutr 18:521–531. https://doi.org/10.1111/j.1365-2095.2011.00923.x
Koh HG, Kang NK, Kim EK et al (2018) Advanced multigene expression system for Nannochloropsis salina using 2A self-cleaving peptides. J Biotechnol 278:39–47. https://doi.org/10.1016/J.JBIOTEC.2018.04.017
Koven W, Barr Y, Lutzky S et al (2001) The effect of dietary arachidonic acid (20:4n−6) on growth, survival and resistance to handling stress in gilthead seabream (Sparus aurata) larvae. Aquaculture 193:107–122. https://doi.org/10.1016/S0044-8486(00)00479-8
Krienitz L, Wirth M (2006) The high content of polyunsaturated fatty acids in Nannochloropsis limnetica (Eustigmatophyceae) and its implication for food web interactions, freshwater aquaculture and biotechnology. Limnologica 36:204–210. https://doi.org/10.1016/j.limno.2006.05.002
Kyle DJ (2001) The large-scale production and use of a single-cell oil highly enriched in docosahexaenoic acid. In: Omega-3 fat acids, pp 92–107. https://doi.org/10.1021/bk-2001-0788.ch008
Lee Chang KJ, Nichols CM, Blackburn SI et al (2014) Comparison of thraustochytrids Aurantiochytrium sp., Schizochytrium sp., Thraustochytrium sp., and Ulkenia sp. for production of biodiesel, long-chain omega-3 oils, and exopolysaccharide. Mar Biotechnol 16:396–411. https://doi.org/10.1007/s10126-014-9560-5
León-Saiki GM, Remmers IM, Martens DE et al (2017) The role of starch as transient energy buffer in synchronized microalgal growth in Acutodesmus obliquus. Algal Res 25:160–167. https://doi.org/10.1016/j.algal.2017.05.018
Leu S, Boussiba S (2014) Advances in the production of high-value products by microalgae. Ind Biotechnol 10:169–183. https://doi.org/10.1089/ind.2013.0039
Levering J, Broddrick J, Dupont CL et al (2016) Genome-scale model reveals metabolic basis of biomass partitioning in a model diatom. PLoS One 11:e0155038. https://doi.org/10.1371/journal.pone.0155038
Levitan O, Dinamarca J, Zelzion E et al (2015a) An RNA interference knock-down of nitrate reductase enhances lipid biosynthesis in the diatom Phaeodactylum tricornutum. Plant J 84:963–973. https://doi.org/10.1111/tpj.13052
Levitan O, Dinamarca J, Zelzion E et al (2015b) Remodeling of intermediate metabolism in the diatom Phaeodactylum tricornutum under nitrogen stress. Proc Natl Acad Sci 112:412–417. https://doi.org/10.1073/pnas.1419818112
Leyland B, Leu S, Boussiba S, Voglmayr H (2017) Are Thraustochytrids algae? Fungal Biol 121:835–840. https://doi.org/10.1016/j.funbio.2017.07.006
Li Z, Bock R (2018) Replication of bacterial plasmids in the nucleus of the red alga Porphyridium purpureum. Nat Commun 9:3451. https://doi.org/10.1038/s41467-018-05651-1
Li X, Marella TK, Tao L et al (2017a) Optimization of growth conditions and fatty acid analysis for three freshwater diatom isolates. Phycol Res 65:177–187. https://doi.org/10.1111/pre.12174
Li X, Pei G, Liu L et al (2017b) Metabolomic analysis and lipid accumulation in a glucose tolerant Crypthecodinium cohnii strain obtained by adaptive laboratory evolution. Bioresour Technol 235:87–95. https://doi.org/10.1016/J.BIORTECH.2017.03.049
Liang Y, Maeda Y, Sunaga Y et al (2013) Biosynthesis of polyunsaturated fatty acids in the oleaginous marine diatom Fistulifera sp. strain JPCC DA0580. Mar Drugs 11:5008–5023. https://doi.org/10.3390/md11125008
Liang Y, Maeda Y, Yoshino T et al (2014) Profiling of polar lipids in marine oleaginous diatom Fistulifera solaris JPCC DA0580: prediction of the potential mechanism for eicosapentaenoic acid-incorporation into triacylglycerol. Mar Drugs 12:3218–3230. https://doi.org/10.3390/md12063218
Li-Beisson Y, Shorrosh B, Beisson F et al (2013) Acyl-lipid metabolism. Arab B 11:e0161. https://doi.org/10.1199/tab.0161
Liu C-P, Lin L-P (2005) Morphology and eicosapentaenoic acid production by Monodus subterraneus UTEX 151. Micron 36:545–550. https://doi.org/10.1016/j.micron.2005.05.001
Longworth J, Wu D, Huete-Ortega M et al (2016) Proteome response of Phaeodactylum tricornutum, during lipid accumulation induced by nitrogen depletion. Algal Res 18:213–224. https://doi.org/10.1016/j.algal.2016.06.015
Lopes-Marques M, Kabeya N, Qian Y et al (2018) Retention of fatty acyl desaturase 1 (fads1) in Elopomorpha and Cyclostomata provides novel insights into the evolution of long-chain polyunsaturated fatty acid biosynthesis in vertebrates. BMC Evol Biol 18:157. https://doi.org/10.1186/s12862-018-1271-5
Lupette J, Jaussaud A, Vigor C et al (2018) Non-enzymatic synthesis of bioactive isoprostanoids in the diatom phaeodactylum following oxidative stress. Plant Physiol 178:1344–1357. https://doi.org/10.1104/PP.18.00925
Ma Y-H, Wang X, Niu Y-F et al (2014) Antisense knockdown of pyruvate dehydrogenase kinase promotes the neutral lipid accumulation in the diatom Phaeodactylum tricornutum. Microb Cell Fact 13:100. https://doi.org/10.1186/s12934-014-0100-9
Ma XN, Chen TP, Yang B et al (2016) Lipid production from Nannochloropsis. Mar Drugs 14. https://doi.org/10.3390/md14040061
Ma H, Jin M, Zhu T et al (2018) Effect of dietary arachidonic acid levels on growth performance, fatty acid profiles and lipid metabolism of juvenile yellow catfish (Pelteobagrus fulvidraco). Aquaculture 486:31–41. https://doi.org/10.1016/J.AQUACULTURE.2017.11.055
Martins DA, Custódio L, Barreira L et al (2013) Alternative sources of n-3 long-chain polyunsaturated fatty acids in marine microalgae. Mar Drugs 11(7):2259–2281. https://doi.org/10.3390/md11072259
Matsuda T, Sakaguchi K, Hamaguchi R et al (2012) Analysis of Δ12-fatty acid desaturase function revealed that two distinct pathways are active for the synthesis of PUFAs in T. aureum ATCC 34304. J Lipid Res 53:1210–1222. https://doi.org/10.1194/jlr.M024935
Mazzocchi A, D’Oria V, Cosmi V et al (2018) The role of lipids in human milk and infant formulae. Nutrients 10:567. https://doi.org/10.3390/nu10050567
Meesapyodsuk D, Qiu X (2012) The front-end desaturase: structure, function, evolution and biotechnological use. Lipids 47:227–237. https://doi.org/10.1007/s11745-011-3617-2
Meng Y, Jiang J, Wang H et al (2015) The characteristics of TAG and EPA accumulation in Nannochloropsis oceanica IMET1 under different nitrogen supply regimes. Bioresour Technol 179:483–489. https://doi.org/10.1016/j.biortech.2014.12.012
Metz JG, Roessler P, Facciotti D et al (2001) Production of polyunsaturated fatty acids by polyketide synthases in both prokaryotes and eukaryotes. Science 293:290–293. https://doi.org/10.1126/science.1059593
Metz JG, Kuner J, Rosenzweig B et al (2009) Biochemical characterization of polyunsaturated fatty acid synthesis in Schizochytrium: release of the products as free fatty acids. Plant Physiol Biochem 47:472–478. https://doi.org/10.1016/j.plaphy.2009.02.002
Miyagawa-Yamaguchi A, Okami T, Kira N et al (2011) Stable nuclear transformation of the diatom Chaetoceros sp. Phycol Res 59:113–119. https://doi.org/10.1111/j.1440-1835.2011.00607.x
Miyahara M, Aoi M, Inoue-Kashino N et al (2013) Highly efficient transformation of the diatom Phaeodactylum tricornutum by multi-pulse electroporation. Biosci Biotechnol Biochem 77:874–876. https://doi.org/10.1271/bbb.120936
Moog D, Stork S, Reislöhner S et al (2015) In vivo localization studies in the stramenopile alga Nannochloropsis oceanica. Protist 166:161–171. https://doi.org/10.1016/j.protis.2015.01.003
Murakami H, Nobusawa T, Hori K et al (2018) Betaine lipid is crucial for adapting to low temperature and phosphate deficiency in Nannochloropsis. Plant Physiol 177:181–193. https://doi.org/10.1104/pp.17.01573
Muto M, Fukuda Y, Nemoto M et al (2013) Establishment of a genetic transformation system for the marine pennate diatom Fistulifera sp. strain JPCC DA0580-a high triglyceride producer. Mar Biotechnol 15:48–55. https://doi.org/10.1007/s10126-012-9457-0
Nath PRR, Khozin-Goldberg I, Cohen Z et al (2012) Dietary supplementation with the microalgae Parietochloris incisa increases survival and stress resistance in guppy (Poecilia reticulata) fry. Aquac Nutr 18:167–180. https://doi.org/10.1111/j.1365-2095.2011.00885.x
Nayak S, Koven W, Meiri I et al (2017) Dietary arachidonic acid affects immune function and fatty acid composition in cultured rabbitfish Siganus rivulatus. Fish Shellfish Immunol 68:46–53. https://doi.org/10.1016/j.fsi.2017.07.003
Nayak S, Khozin-Goldberg I, Cohen G, Zilberg D (2018) Dietary supplementation with ω6 LC-PUFA-rich algae modulates zebrafish immune function and improves resistance to streptococcal infection. Front Immunol 9:1960. https://doi.org/10.3389/fimmu.2018.01960
Neumann U, Derwenskus F, Gille A et al (2018) Bioavailability and safety of nutrients from the microalgae Chlorella vulgaris, Nannochloropsis oceanica and Phaeodactylum tricornutum in C57BL/6 Mice. Nutrients 10:965. https://doi.org/10.3390/nu10080965
Nguyen HM, Cuiné S, Beyly-Adriano A et al (2013) The green microalga Chlamydomonas reinhardtii has a single ω-3 fatty acid desaturase that localizes to the chloroplast and impacts both plastidic and extraplastidic membrane lipids. Plant Physiol 163:914–928. https://doi.org/10.1104/pp.113.223941
Niu YF, Zhang MH, Li DW et al (2013) Improvement of neutral lipid and polyunsaturated fatty acid biosynthesis by overexpressing a type 2 diacylglycerol acyltransferase in marine diatom Phaeodactylum tricornutum. Mar Drugs 11:4558–4569. https://doi.org/10.3390/md11114558
Niu Y-F, Wang X, Hu D-X et al (2016) Molecular characterization of a glycerol-3-phosphate acyltransferase reveals key features essential for triacylglycerol production in Phaeodactylum tricornutum. Biotechnol Biofuels 9:60. https://doi.org/10.1186/s13068-016-0478-1
Nugent AP (2004) The metabolic syndrome. Nutr Bull 29:36–43
Nymark M, Sharma AK, Sparstad T et al (2016) A CRISPR/Cas9 system adapted for gene editing in marine algae. Sci Rep 6:24951. https://doi.org/10.1038/srep24951
Ohlrogge J, Browse J (1995) Lipid biosynthesis. Plant Cell 7:957–970. https://doi.org/10.1105/tpc.7.7.957
Pal D, Khozin-Goldberg I, Cohen Z, Boussiba S (2011) The effect of light, salinity, and nitrogen availability on lipid production by Nannochloropsis sp. Appl Microbiol Biotechnol 90:1429–1441. https://doi.org/10.1007/s00253-011-3170-1
Pal D, Khozin-Goldberg I, Didi-Cohen S et al (2013) Growth, lipid production and metabolic adjustments in the euryhaline eustigmatophyte Nannochloropsis oceanica CCALA 804 in response to osmotic downshift. Appl Microbiol Biotechnol 97:8291–8306. https://doi.org/10.1007/s00253-013-5092-6
Pal-Nath D, Didi-Cohen S, Shtaida N et al (2017) Improved productivity and oxidative stress tolerance under nitrogen starvation is associated with the ablated Δ5 desaturation in the green microalga Lobosphaera incisa. Algal Res 26:25–38. https://doi.org/10.1016/J.ALGAL.2017.06.026
Pasquet V, Ulmann L, Mimouni V et al (2014) Fatty acids profile and temperature in the cultured marine diatom Odontella aurita. J Appl Phycol 26:2265–2271. https://doi.org/10.1007/s10811-014-0252-3
Patron NJ, Orzaez D, Marillonnet S et al (2015) Standards for plant synthetic biology: a common syntax for exchange of DNA parts. New Phytol 208:13–19. https://doi.org/10.1111/nph.13532
Peng K-T (2014) Delta 5 fatty acid desaturase upregulates the synthesis of polyunsaturated fatty acids in the marine diatom Phaeodactylum tricornutum. J Agric Food Chem 62:8773–8776
Perez-Garcia O, Escalante FME, de Bashan LE, Bashan Y (2011) Heterotrophic cultures of microalgae: metabolism and potential products. Water Res 45:11–36. https://doi.org/10.1016/j.watres.2010.08.037
Perin G, Bellan A, Segalla A et al (2015) Generation of random mutants to improve light-use efficiency of Nannochloropsis gaditana cultures for biofuel production. Biotechnol Biofuels 8:161. https://doi.org/10.1186/s13068-015-0337-5
Perrineau M-M, Zelzion E, Gross J et al (2014) Evolution of salt tolerance in a laboratory reared population of Chlamydomonas reinhardtii. Environ Microbiol 16:1755–1766. https://doi.org/10.1111/1462-2920.12372
Petrie JR, Liu Q, Mackenzie AM et al (2010a) Isolation and characterisation of a high-efficiency desaturase and elongases from microalgae for transgenic LC-PUFA production. Mar Biotechnol 12:430–438. https://doi.org/10.1007/s10126-009-9230-1
Petrie JR, Shrestha P, Mansour MP et al (2010b) Metabolic engineering of omega-3 long-chain polyunsaturated fatty acids in plants using an acyl-CoA Δ6-desaturase with ω3-preference from the marine microalga Micromonas pusilla. Metab Eng 12:233–240. https://doi.org/10.1016/j.ymben.2009.12.001
Poliner E, Pulman JA, Zienkiewicz K et al (2017) A toolkit for Nannochloropsis oceanica CCMP1779 enables gene stacking and genetic engineering of the eicosapentaenoic acid pathway for enhanced long-chain polyunsaturated fatty acid production. Plant Biotechnol J 16:298–309. https://doi.org/10.1111/pbi.12772
Poliner E, Farré EM, Benning C (2018a) Advanced genetic tools enable synthetic biology in the oleaginous microalgae Nannochloropsis sp. Plant Cell Rep 37:1383–1399. https://doi.org/10.1007/s00299-018-2270-0
Poliner E, Takeuchi T, Du ZY et al (2018b) Nontransgenic marker-free gene disruption by an episomal CRISPR system in the oleaginous microalga, Nannochloropsis oceanica CCMP1779. ACS Synth Biol 7:962–968. https://doi.org/10.1021/acssynbio.7b00362
Popko J, Herrfurth C, Feussner K et al (2016) Metabolome analysis reveals betaine lipids as major source for triglyceride formation, and the accumulation of sedoheptulose during nitrogen-starvation of Phaeodactylum tricornutum. PLoS One 11(10):e0164673. https://doi.org/10.1371/journal.pone.0164673
Poulsen N, Kröger N (2005) A new molecular tool for transgenic diatoms: control of mRNA and protein biosynthesis by an inducible promoter-terminator cassette. FEBS J 272:3413–3423. https://doi.org/10.1111/j.1742-4658.2005.04760.x
Prasad B, Vadakedath N, Jeong HJ et al (2014) Agrobacterium tumefaciens-mediated genetic transformation of haptophytes (Isochrysis species). Appl Microbiol Biotechnol 98:8629–8639. https://doi.org/10.1007/s00253-014-5900-7
Prasad B, Lein W, Thiyam G et al (2018) Stable nuclear transformation of rhodophyte species Porphyridium purpureum: advanced molecular tools and an optimized method. Photosynth Res 140:173–188. https://doi.org/10.1007/s11120-018-0587-8
Qi B, Fraser TCM, Bleakley CL et al (2003) The variant “his-box” of the C18-Delta9-PUFA-specific elongase IgASE1 from Isochrysis galbana is essential for optimum enzyme activity. FEBS Lett 547:137–139. https://doi.org/10.1016/S0014-5793(03)00676-8
Qi BX, Fraser T, Mugford S et al (2004) Production of very long chain polyunsaturated omega-3 and omega-6 fatty acids in plants. Nat Biotechnol 22:739–745. https://doi.org/10.1038/Nbt972
Qiu X (2003) Biosynthesis of docosahexaenoic acid (DHA, 22:6-4, 7,10,13,16,19): two distinct pathways. Prostaglandins Leukot Essent Fat Acids 68:181–186. https://doi.org/10.1016/S0952-3278(02)00268-5
Radakovits R, Eduafo PM, Posewitz MC (2011) Genetic engineering of fatty acid chain length in Phaeodactylum tricornutum. Metab Eng 13:89–95. https://doi.org/10.1016/j.ymben.2010.10.003
Ratledge C, Kanagachandran K, Anderson AJ et al (2001) Production of docosahexaenoic acid by Crypthecodinium cohnii grown in a pH-auxostat culture with acetic acid as principal carbon source. Lipids 36:1241–1246. https://doi.org/10.1007/s11745-001-0838-x
Recchiuti A, Serhan CN (2012) Pro-resolving lipid mediators (SPMs) and their actions in regulating miRNA in novel resolution circuits in inflammation. Front Immunol 3:298. https://doi.org/10.3389/fimmu.2012.00298
Remmers IM, D’Adamo S, Martens DE et al (2018) Orchestration of transcriptome, proteome and metabolome in the diatom Phaeodactylum tricornutum during nitrogen limitation. Algal Res 35:33–49. https://doi.org/10.1016/j.algal.2018.08.012
Řezanka T, Kolouchová I, Nedbalová L, Sigler K (2018) Enantiomeric separation of triacylglycerols containing very long chain fatty acids. J Chromatogr A 1557:9–19. https://doi.org/10.1016/j.chroma.2018.04.064
Rodolfi L, Biondi N, Guccione A et al (2017) Oil and eicosapentaenoic acid production by the diatom Phaeodactylum tricornutum cultivated outdoors in Green Wall Panel (GWP®) reactors. Biotechnol Bioeng 114:2204–2210. https://doi.org/10.1002/bit.26353
Roessler PG (1987) UDP-glucose pyrophosphorylase activity in the diatom Cyclotella cryptica pathway of chrysolaminarin synthesis. J Phycol 23:494–498. https://doi.org/10.1111/j.1529-8817.1987.tb02536.x
Ruiz-Lopez N, Haslam RP, Usher S et al (2015) An alternative pathway for the effective production of the omega-3 long-chain polyunsaturates EPA and ETA in transgenic oilseeds. Plant Biotechnol J 13:1–12. https://doi.org/10.1111/pbi.12328
Russo MT, Aiese Cigliano R, Sanseverino W, Ferrante MI (2018) Assessment of genomic changes in a CRISPR/Cas9 Phaeodactylum tricornutum mutant through whole genome resequencing. Peer J 6:e5507. https://doi.org/10.7717/peerj.5507
Ryckebosch E, Bruneel C, Muylaert K, Foubert I (2012) Microalgae as an alternative source of omega-3 long chain polyunsaturated fatty acids. Lipid Technol 24:128–130. https://doi.org/10.1002/lite.201200197
Safafar H, Hass M, Møller P et al (2016) High-EPA biomass from Nannochloropsis salina cultivated in a flat-panel photo-bioreactor on a process water-enriched growth medium. Mar Drugs 14:144. https://doi.org/10.3390/md14080144
Sarker PK, Kapuscinski AR, Bae AY et al (2018) Towards sustainable aquafeeds: evaluating substitution of fishmeal with lipid-extracted microalgal co-product (Nannochloropsis oculata) in diets of juvenile Nile tilapia (Oreochromis niloticus). PLoS One 13:e0201315. https://doi.org/10.1371/journal.pone.0201315
Sayanova O, Napier JA (2016) Metabolic engineering of microalgae for sustainable production of omega-3 long chain polyunsaturated fatty acids. Curr Biotechnol 5:198–212. https://doi.org/10.2174/2211550105666160223214550
Sayanova O, Smith MA, Lapinskas P et al (1997) Expression of a borage desaturase cDNA containing an N-terminal cytochrome b5 domain results in the accumulation of high levels of delta6-desaturated fatty acids in transgenic tobacco. Proc Natl Acad Sci USA 94:4211–4216. https://doi.org/10.1073/PNAS.94.8.4211
Sayanova O, Haslam RP, Calerón MV et al (2011) Identification and functional characterisation of genes encoding the omega-3 polyunsaturated fatty acid biosynthetic pathway from the coccolithophore Emiliania huxleyi. Phytochemistry 72:594–600. https://doi.org/10.1016/j.phytochem.2011.01.022
Sayanova O, Ruiz-Lopez N, Haslam RP, Napier JA (2012) The role of Δ6-desaturase acyl-carrier specificity in the efficient synthesis of long-chain polyunsaturated fatty acids in transgenic plants. Plant Biotechnol J 10:195–206. https://doi.org/10.1111/j.1467-7652.2011.00653.x
Sayanova O, Mimouni V, Ulmann L et al (2017) Modulation of lipid biosynthesis by stress in diatoms. Philos Trans R Soc B Biol Sci 372:20160407. https://doi.org/10.1098/rstb.2016.0407
Scaife MA, Nguyen GTDT, Rico J et al (2015) Establishing Chlamydomonas reinhardtii as an industrial biotechnology host. Plant J 82:532–546. https://doi.org/10.1111/tpj.12781
Schneider JC, Roessler P (1994) Radiolabeling studies of lipids and fatty acids in Nannochloropsis (Eustigmatophyceae), an oleaginous marine alga. J Phycol 30:594–598
Schneider JC, Livne A, Sukenik A, Roessler PG (1995) A mutant of Nannochloropsis deficient in eicosapentaenoic acid production. Phytochemistry 40:807–814. https://doi.org/10.1016/0031-9422(95)00365-E
Schwartz AS, Brown R, Ajjawi I et al (2018) Complete genome sequence of the model oleaginous alga Nannochloropsis gaditana CCMP1894. Genome Announc 6. https://doi.org/10.1128/genomeA.01448-17
Serhan CN (2014) Pro-resolving lipid mediators are leads for resolution physiology. Nature 510:92–101. https://doi.org/10.1038/nature13479
Shanklin J, Guy JE, Mishra G, Lindqvist Y (2009) Desaturases: emerging models for understanding functional diversification of diiron-containing enzymes. J Biol Chem 284:18559–18563. https://doi.org/10.1074/jbc.R900009200
Shepherd CJ, Jackson AJ (2013) Global fishmeal and fish-oil supply: inputs, outputs and markets. J Fish Biol 83:1046–1066. https://doi.org/10.1111/jfb.12224
Shi T, Yu A, Li M et al (2012) Identification of a novel C22-Δ4-producing docosahexaenoic acid (DHA) specific polyunsaturated fatty acid desaturase gene from Isochrysis galbana and its expression in Saccharomyces cerevisiae. Biotechnol Lett 34:2265–2274. https://doi.org/10.1007/s10529-012-1028-y
Shi H, Wu R, Zheng Y, Yue X (2018) Molecular mechanisms underlying catalytic activity of delta 6 desaturase from Glossomastix chrysoplasta and Thalassiosira pseudonana. J Lipid Res 59:79–88. https://doi.org/10.1194/jlr.M079806
Shields RJ, Lupatsch I (2012) Algae for aquaculture and animal feeds. Tech – Theor Prax 21:23–37
Shockey J, Regmi A, Cotton K et al (2016) Identification of Arabidopsis GPAT9 (At5g60620) as an essential gene involved in triacylglycerol biosynthesis. Plant Physiol 170:163–179. https://doi.org/10.1104/pp.15.01563
Shrestha RP, Hildebrand M (2017) Development of a silicon limitation inducible expression system for recombinant protein production in the centric diatoms Thalassiosira pseudonana and Cyclotella cryptica. Microb Cell Factories 16:145. https://doi.org/10.1186/s12934-017-0760-3
Simopoulos AP (2002) The importance of the ratio of omega-6/omega-3 essential fatty acids. Biomed Pharmacother 56:365–379
Simopoulos AP (2008) The importance of the omega-6/omega-3 fatty acid ratio in cardiovascular disease and other chronic diseases. Exp Biol Med (Maywood) 233:674–688. https://doi.org/10.3181/0711-MR-311
Singer SD, Chen G, Mietkiewska E et al (2016) Arabidopsis GPAT9 contributes to synthesis of intracellular glycerolipids but not surface lipids. J Exp Bot 67:4627–4638. https://doi.org/10.1093/jxb/erw242
Skrede A, Mydland L, Ahlstrøm Ø et al (2011) Evaluation of microalgae as sources of digestible nutrients for monogastric animals. J Anim Feed Sci 20:131–142. https://doi.org/10.22358/jafs/66164/2011
Slattery SS, Diamond A, Wang H et al (2018) An expanded plasmid-based genetic toolbox enables Cas9 genome editing and stable maintenance of synthetic pathways in Phaeodactylum tricornutum. ACS Synth Biol 7:328–338. https://doi.org/10.1021/acssynbio.7b00191
Smith SR, Gillard JTF, Kustka AB et al (2016) Transcriptional orchestration of the global cellular response of a model pennate diatom to diel light cycling under iron limitation. PLoS Genet 12:e1006490. https://doi.org/10.1371/journal.pgen.1006490
Solovchenko AE, Khozin-Goldberg I, Didi-Cohen S et al (2008) Effects of light intensity and nitrogen starvation on growth, total fatty acids and arachidonic acid in the green microalga Parietochloris incisa. J Appl Phycol 20:245–251. https://doi.org/10.1007/s10811-007-9233-0
Spicer A, Molnar A (2018) Gene editing of microalgae: scientific progress and regulatory challenges in Europe. Biology (Basel) 7:21. https://doi.org/10.3390/biology7010021
Sprague M, Betancor MB, Tocher DR (2017) Microbial and genetically engineered oils as replacements for fish oil in aquaculture feeds. Biotechnol Lett 39:1599–1609. https://doi.org/10.1007/s10529-017-2402-6
Starkenburg SR, Kwon KJ, Jha RK et al (2014) A pangenomic analysis of the Nannochloropsis organellar genomes reveals novel genetic variations in key metabolic genes. BMC Genomics 15:212. https://doi.org/10.1186/1471-2164-15-212
Steinrücken P, Prestegard SK, de Vree JH et al (2018) Comparing EPA production and fatty acid profiles of three Phaeodactylum tricornutum strains under western Norwegian climate conditions. Algal Res 30:11–22. https://doi.org/10.1016/j.algal.2017.12.001
Su Y, Yi Z, Björnsdóttir SH et al (2018) Adaptive laboratory evolution for enhanced carotenoid production in microalgae. Methods Mol Biol 1852:117–126
Sukenik A (1991) Ecophysiological considerations in the optimization of eicosapentaenoic acid production by Nannochloropsis sp. (Eustigmatophyceae). Bioresourc Technol 35:263–269
Sukenik A, Beardall J, Kromkamp JC et al (2009) Photosynthetic performance of outdoor Nannochloropsis mass cultures under a wide range of environmental conditions. Aquat Microb Ecol 56:297–308. https://doi.org/10.3354/ame01309
Sun X-M, Ren L-J, Ji X-J et al (2016) Adaptive evolution of Schizochytrium sp. by continuous high oxygen stimulations to enhance docosahexaenoic acid synthesis. Bioresour Technol 211:374–381. https://doi.org/10.1016/J.BIORTECH.2016.03.093
Sun X-M, Ren L-J, Bi Z-Q et al (2018) Adaptive evolution of microalgae Schizochytrium sp. under high salinity stress to alleviate oxidative damage and improve lipid biosynthesis. Bioresour Technol 267:438–444. https://doi.org/10.1016/J.BIORTECH.2018.07.079
Szyjka SJ, Mandal S, Schoepp NG et al (2017) Evaluation of phenotype stability and ecological risk of a genetically engineered alga in open pond production. Algal Res 24:378–386. https://doi.org/10.1016/J.ALGAL.2017.04.006
Tanaka T, Maeda Y, Veluchamy A et al (2015) Oil accumulation by the oleaginous diatom Fistulifera solaris as revealed by the genome and transcriptome. Plant Cell 27:162–176. https://doi.org/10.1105/tpc.114.135194
Taoka Y, Nagano N, Okita Y et al (2009) Influences of culture temperature on the growth, lipid content and fatty acid composition of Aurantiochytrium sp. Strain mh0186. Mar Biotechnol (NY) 11:368–374. https://doi.org/10.1007/s10126-008-9151-4
Tejera N, Vauzour D, Betancor MB et al (2016) A transgenic Camelina sativa seed oil effectively replaces fish oil as a dietary source of eicosapentaenoic acid in mice. J Nutr 146:227–235. https://doi.org/10.3945/jn.115.223941.227
Tirichine L, Bowler C (2011) Decoding algal genomes: tracing back the history of photosynthetic life on Earth. Plant J 66:45–57. https://doi.org/10.1111/j.1365-313X.2011.04540.x
Tocher DR (2015) Omega-3 long-chain polyunsaturated fatty acids and aquaculture in perspective. Aquaculture 449:94–107. https://doi.org/10.1016/j.aquaculture.2015.01.010
Tocher DR, Glencross BD (2015) Lipids and fatty acids. In: Lee C-S, Lim C, Webster C, Gatlin DM (eds) Dietary nutrients, additives, and fish health. Wiley, Hoboken, pp 47–94
Tocher D, Betancor M, Sprague M et al (2019) Omega-3 long-chain polyunsaturated fatty acids, EPA and DHA: bridging the gap between supply and demand. Nutrients 11:89. https://doi.org/10.3390/nu11010089
Tonon T, Harvey D, Larson TR, Graham IA (2002) Long chain polyunsaturated fatty acid production and partitioning to triacylglycerols in four microalgae. Phytochemistry 61:15–24
Tonon T, Qing R, Harvey D et al (2005a) Identification of a long-chain polyunsaturated fatty acid acyl-coenzyme A synthetase from the diatom Thalassiosira pseudonana. Plant Physiol 138:402–408. https://doi.org/10.1104/pp.104.054528.activity
Tonon T, Sayanova O, Michaelson LV et al (2005b) Fatty acid desaturases from the microalga Thalassiosira pseudonana. FEBS J 272:3401–3412. https://doi.org/10.1111/j.1742-4658.2005.04755.x
Trentacoste EM, Shrestha RP, Smith SR et al (2013) Metabolic engineering of lipid catabolism increases microalgal lipid accumulation without compromising growth. Proc Natl Acad Sci USA 110:1–6. https://doi.org/10.1073/pnas.1309299110
Usher S, Han L, Haslam RP et al (2017) Tailoring seed oil composition in the real world: optimising omega-3 long chain polyunsaturated fatty acid accumulation in transgenic Camelina sativa. Sci Rep 7:6570. https://doi.org/10.1038/s41598-017-06838-0
Vaezi R, Napier J, Sayanova O et al (2013) Identification and functional characterization of genes encoding omega-3 polyunsaturated fatty acid biosynthetic activities from unicellular microalgae. Mar Drugs 11:5116–5129. https://doi.org/10.3390/md11125116
Vazquez-Vilar M, Orzaez D, Patron N (2018) DNA assembly standards: setting the low-level programming code for plant biotechnology. Plant Sci 273:33–41. https://doi.org/10.1016/J.PLANTSCI.2018.02.024
Verruto J, Francis K, Wang Y et al (2018) Unrestrained markerless trait stacking in Nannochloropsis gaditana through combined genome editing and marker recycling technologies. Proc Natl Acad Sci 115:E7015–E7022. https://doi.org/10.1073/pnas.1718193115
Vieler A, Wu G, Tsai C-H et al (2012) Genome, functional gene annotation, and nuclear transformation of the heterokont oleaginous alga Nannochloropsis oceanica CCMP1779. PLoS Genet 8:e1003064. https://doi.org/10.1371/journal.pgen.1003064
Vigani M, Parisi C, Rodr Iguez-Cerezo E et al (2015) Food and feed products from micro-algae: market opportunities and challenges for the EU. Trends Food Sci Technol 42:81–92. https://doi.org/10.1016/j.tifs.2014.12.004
Wallis JG, Browse J (1999) The Delta8-desaturase of Euglena gracilis: an alternate pathway for synthesis of 20-carbon polyunsaturated fatty acids. Arch Biochem Biophys 365:307–316. https://doi.org/10.1006/abbi.1999.1167
Wang D, Ning K, Li J et al (2014) Nannochloropsis genomes reveal evolution of microalgal oleaginous traits. PLoS Genet 10:e1004094. https://doi.org/10.1371/journal.pgen.1004094
Wang X, Liu Y-H, Wei W et al (2017) Enrichment of long-chain polyunsaturated fatty acids by coordinated expression of multiple metabolic nodes in the oleaginous microalga Phaeodactylum tricornutum. J Agric Food Chem 65(35):7713–7720. https://doi.org/10.1021/acs.jafc.7b02397
Wang X, Dong H-P, Wei W et al (2018) Dual expression of plastidial GPAT1 and LPAT1 regulates triacylglycerol production and the fatty acid profile in Phaeodactylum tricornutum. Biotechnol Biofuels 11:318. https://doi.org/10.1186/s13068-018-1317-3
Wase N, Black P, DiRusso C (2018) Innovations in improving lipid production: algal chemical genetics. Prog Lipid Res 71:101–123. https://doi.org/10.1016/J.PLIPRES.2018.07.001
Wen Z-Y, Chen F (2003) Heterotrophic production of eicosapentaenoic acid by microalgae. Biotechnol Adv 21:273–294. https://doi.org/10.1016/S0734-9750(03)00051-X
Wu G, Truksa M, Datla N et al (2005) Stepwise engineering to produce high yields of very long-chain polyunsaturated fatty acids in plants. Nat Biotechnol 23:1013–1017. https://doi.org/10.1038/nbt1107
Xie D, Jackson EN, Zhu Q (2015) Sustainable source of omega-3 eicosapentaenoic acid from metabolically engineered Yarrowia lipolytica: from fundamental research to commercial production. Appl Microbiol Biotechnol 99:1599–1610. https://doi.org/10.1007/s00253-014-6318-y
Xue Z, Sharpe PL, Hong S-P et al (2013) Production of omega-3 eicosapentaenoic acid by metabolic engineering of Yarrowia lipolytica. Nat Biotechnol 31:734–740. https://doi.org/10.1038/nbt.2622
Yokochi T, Honda D, Higashihara T, Nakahara T (1998) Optimization of docosahexaenoic acid production by Schizochytrium limacinum SR21. Appl Microbiol Biotechnol 49:72–76. https://doi.org/10.1007/s002530051139
Yongmanitchai W, Ward OP (1991) Growth of and omega-3 fatty acid production by Phaeodactylum tricornutum under different culture conditions. Appl Environ Microbiol 57:419–425
Yu R, Yamada A, Watanabe K et al (2000) Production of eicosapentaenoic acid by a recombinant marine cyanobacterium, Synechococcus sp. Lipids 35:1061–1064. https://doi.org/10.1007/s11745-000-0619-6
Zaslavskaia LA, Lippmeier JC, Kroth PG et al (2000) Transformation of the diatom Phaeodactylum tricornutum with a variety of selectable marker and reporter genes. J Phycol 386:379–386. https://doi.org/10.1046/j.1529-8817.2000.99164.x
Zhou X-R, Robert SS, Petrie JR et al (2007) Isolation and characterization of genes from the marine microalga Pavlova salina encoding three front-end desaturases involved in docosahexaenoic acid biosynthesis. Phytochemistry 68:785–796. https://doi.org/10.1016/j.phytochem.2006.12.016
Zhu BH, Shi HP, Yang GP et al (2016) Silencing UDP-glucose pyrophosphorylase gene in Phaeodactylum tricornutum affects carbon allocation. New Biotechnol 33:237–244. https://doi.org/10.1016/j.nbt.2015.06.003
Zhukova NV, Aizdaicher NA (1995) Fatty acid composition of 15 species of marine microalgae. Phytochemistry 39:351–356. https://doi.org/10.1016/0031-9422(94)00913-E
Zittelli GC, Lavista F, Bastianini A et al (1999) Production of eicosapentaenoic acid by Nannochloropsis sp. cultures in outdoor tubular photobioreactors. J Biotechnol 70:299–312
Zorin B, Grundman O, Khozin-Goldberg I et al (2014) Development of a nuclear transformation system for oleaginous green alga Lobosphaera (Parietochloris) incisa and genetic complementation of a mutant strain, deficient in arachidonic acid biosynthesis. PLoS One 9:e105223. https://doi.org/10.1371/journal.pone.0105223
Zorin B, Pal-Nath D, Lukyanov A et al (2017) Arachidonic acid is important for efficient use of light by the microalga Lobosphaera incisa under chilling stress. Biochim Biophys Acta Mol Cell Biol Lipids 1862:853–868. https://doi.org/10.1016/j.bbalip.2017.04.008
Acknowledgements
I.K.G. acknowledges support by a grant from the Ministry of Science, Technology and Space, Israel. Rothamsted Research receives grant-aided support from the Biotechnology and Biological Sciences Research Council (BBSRC), UK. This project was supported by BBSRC grant BB/L002957/1.
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Khozin-Goldberg, I., Sayanova, O. (2019). Metabolic Engineering and Synthetic Biology Approaches to Enhancing Production of Long-Chain Polyunsaturated Fatty Acids in Microalgae. In: Hallmann, A., Rampelotto, P. (eds) Grand Challenges in Algae Biotechnology. Grand Challenges in Biology and Biotechnology. Springer, Cham. https://doi.org/10.1007/978-3-030-25233-5_7
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