Research progress and prospects in lipid metabolic engineering of eukaryotic microalgae

SUN Han; LIU Jin

doi:10.12211/2096-8280.2023-044

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Research progress and prospects in lipid metabolic engineering of eukaryotic microalgae

Invited Review | 更新时间：2025-03-20

- Research progress and prospects in lipid metabolic engineering of eukaryotic microalgae
- Synthetic Biology Journal Vol. 4, Issue 6, Pages: 1140-1160(2023)
- 作者机构：
  
  1.深圳大学高等研究院，广东深圳 518060
  2.北京大学工学院，北京 100871
- 作者简介：
- 基金信息：
- DOI：10.12211/2096-8280.2023-044
  CLC： Q819
- Received：29 June 2023，
  
  Revised：2023-08-06，
  
  Published：31 December 2023
- 稿件说明：
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孙翰，刘进. 真核微藻脂质代谢工程的研究进展和展望［J］. 合成生物学， 2023， 4（6）： 1140-1160

SUN Han， LIU Jin. Research progress and prospects in lipid metabolic engineering of eukaryotic microalgae［J］. Synthetic Biology Journal， 2023， 4（6）： 1140-1160
孙翰，刘进. 真核微藻脂质代谢工程的研究进展和展望［J］. 合成生物学， 2023， 4（6）： 1140-1160 DOI： 10.12211/2096-8280.2023-044.

SUN Han， LIU Jin. Research progress and prospects in lipid metabolic engineering of eukaryotic microalgae［J］. Synthetic Biology Journal， 2023， 4（6）： 1140-1160 DOI： 10.12211/2096-8280.2023-044.

摘要

真核微藻作为一类重要的生物资源，脂质含量高，广泛应用于能源、化工和食品等领域。然而，真核微藻生物能源成本偏高，其产业化应用仍然面临着一系列挑战。通过代谢工程手段改造微藻，促进脂质的合成与积累，可提高微藻脂质生产的经济可行性。本文介绍真核微藻脂质代谢途径和关键酶基因，并总结了不同培养条件下代谢途径相关基因在转录水平上的变化。还探讨通过代谢工程调控脂质合成相关酶、转录因子和竞争途径等方法，以提高微藻脂质含量和调整脂肪酸组成。通过基因组学、转录组学和蛋白质组学数据的整合分析可揭示脂质代谢中的关键节点和主效调控因子，有助于确定代谢工程的潜在目标。此外，基因工具和基因编辑技术的开发和拓展可显著提高转化效率，实现对微藻底盘细胞的精准改造。通过重塑能量和碳代谢途径，可设计优化微藻脂质生物合成过程。在微藻遗传工具、基因编辑技术、代谢通路调控和产业化等方面的进一步研究和探索对于推动微藻脂质工程的研究和发展具有重要意义。

Abstract

Microalgae represent a diverse group of photosynthetic organisms that are widely found in various ecosystems on the Earth. They play a crucial role in carbon dioxide bio-fixation. Apart from their efficient growth through photosynthesis

many microalgae can also grow robustly under heterotrophic and mixotrophic conditions for high biomass production. Due to their high lipid content and the presence of diverse fatty acid and lipid species

microalgae have a wide range of applications in industries of energy

chemicals

and food. However

the high production cost associated with microalgae-based bioenergy poses a significant challenge for large-scale implementation. To overcome this

there is a growing interest in engineering microalgae to enhance lipid biosynthesis and accumulation

which holds promise for improving the economic feasibility of microalgal lipid production. This requires a better understanding of lipid metabolism and regulation in microalgae. This article provides an overview of recent advances in the elucidation of lipid metabolic pathways

the roles of key enzyme genes involved in lipid metabolism

and the transcriptional regulation of lipid metabolic pathways under different cultivation conditions in eukaryotic microalgae. It also summarizes strategies for metabolic engineering aiming for manipulating lipid biosynthesis-related enzymes

transcription factors

and competing pathways to increase lipid content and/or modify fatty acid composition in microalgae. Integrated analysis of genomics

transcriptomics

and proteomics data can help identify crucial nodes and key regulators in lipid metabolism

facilitating the identification of potential targets for metabolic engineering. Furthermore

the rapid development of genetic tools and gene editing technologies has significantly improved transformation efficiency and enabled precise gene modification

providing a foundation for genetic engineering of microalgae. By reshaping energy and carbon metabolic pathways

it becomes possible to design and optimize lipid biosynthesis processes in microalgae for a better production. Further research and exploration in genetic tools

gene editing technologies

metabolic pathway regulation

and large-scale implementation are of utmost importance for driving the research and development of microalgal lipid engineering.

关键词

Keywords

references

KATIYAR R , GURJAR B R , BISWAS S , et al . Microalgae: an emerging source of energy based bio-products and a solution for environmental issues [J ] . Renewable and Sustainable Energy Reviews , 2017 , 72 : 1083 - 1093 .

CARNEIRO M L N M , PRADELLE F , BRAGA S L , et al . Potential of biofuels from algae: comparison with fossil fuels, ethanol and biodiesel in Europe and Brazil through life cycle assessment (LCA) [J ] . Renewable and Sustainable Energy Reviews , 2017 , 73 : 632 - 653 .

HEPBURN C , ADLEN E , BEDDINGTON J , et al . The technological and economic prospects for CO 2 utilization and removal [J ] . Nature , 2019 , 575 ( 7781 ): 87 - 97 .

TZACHOR A , RICHARDS C E , HOLT L . Future foods for risk-resilient diets [J ] . Nature Food , 2021 , 2 ( 5 ): 326 - 329 .

ADESANYA V O , CADENA E , SCOTT S A , et al . Life cycle assessment on microalgal biodiesel production using a hybrid cultivation system [J ] . Bioresource Technology , 2014 , 163 : 343 - 355 .

KUMAR K , MISHRA S K , SHRIVASTAV A , et al . Recent trends in the mass cultivation of algae in raceway ponds [J ] . Renewable and Sustainable Energy Reviews , 2015 , 51 : 875 - 885 .

TIBOCHA-BONILLA J D , ZUÑIGA C , GODOY-SILVA R D , et al . Advances in metabolic modeling of oleaginous microalgae [J ] . Biotechnology for Biofuels , 2018 , 11 ( 1 ): 1241 .

LI J , ZHU D L , NIU J F , et al . An economic assessment of astaxanthin production by large scale cultivation of Haematococcus pluvialis [J ] . Biotechnology Advances , 2011 , 29 ( 6 ): 568 - 574 .

RUIZ J , OLIVIERI G , DE VREE J , et al . Towards industrial products from microalgae [J ] . Energy & Environmental Science , 2016 , 9 ( 10 ): 3036 - 3043 .

MASRI M A , GARBE D , MEHLMER N , et al . A sustainable, high-performance process for the economic production of waste-free microbial oils that can replace plant-based equivalents [J ] . Energy & Environmental Science , 2019 , 12 ( 9 ): 2717 - 2732 .

SUN H , MAO X M , WU T , et al . Novel insight of carotenoid and lipid biosynthesis and their roles in storage carbon metabolism in Chlamydomonas reinhardtii [J ] . Bioresource Technology , 2018 , 263 : 450 - 457 .

LAURENS L M L , MARKHAM J , TEMPLETON D W , et al . Development of algae biorefinery concepts for biofuels and bioproducts; a perspective on process-compatible products and their impact on cost-reduction [J ] . Energy & Environmental Science , 2017 , 10 ( 8 ): 1716 - 1738 .

ROTH M S , COKUS S J , GALLAHER S D , et al . Chromosome-level genome assembly and transcriptome of the green alga Chromochloris zofingiensis illuminates astaxanthin production [J ] . Proceedings of the National Academy of Sciences of the United States of America , 2017 , 114 ( 21 ): E4296 - E4305 .

BLANC-MATHIEU R , SANCHEZ-FERANDIN S , EYRE-WALKER A , et al . Organellar inheritance in the green lineage: i nsights from Ostreococcus tauri [J ] . Genome Biology and Evolution , 2013 , 5 ( 8 ): 1503 - 1511 .

WEISS T L , JOHNSTON J S , FUJISAWA K , et al . Genome size and phylogenetic analysis of the A and L races of Botryococcus braunii [J ] . Journal of Applied Phycology , 2011 , 23 ( 5 ): 833 - 839 .

MERCHANT S S , PROCHNIK S E , VALLON O , et al . The Chlamydomonas genome reveals the evolution of key animal and plant functions [J ] . Science , 2007 , 318 ( 5848 ): 245 - 250 .

FAN J H , NING K , ZENG X W , et al . Genomic foundation of starch-to-lipid switch in oleaginous Chlorella spp [J ] . Plant Physiology , 2015 , 169 ( 4 ): 2444 - 2461 .

GUARNIERI M T , BOORE J L , HENARD C A , et al . LDKB00000000.1 Chlorella vulgaris [DB/OL ] . NCBI , 2015 [ 2023-06-01 ] . https://www.ncbi.nlm.nih.gov/Traces/wgs/LDKB01 https://www.ncbi.nlm.nih.gov/Traces/wgs/LDKB01 .

BROUARD J S , OTIS C , LEMIEUX C , et al . The exceptionally large chloroplast genome of the green alga Floydiella terrestris illuminates the evolutionary history of the Chlorophyceae [J ] . Genome Biology and Evolution , 2010 , 2 : 240 - 256 .

PALENIK B , GRIMWOOD J , AERTS A , et al . The tiny eukaryote Ostreococcus provides genomic insights into the paradox of plankton speciation [J ] . Proceedings of the National Academy of Sciences of the United States of America , 2007 , 104 ( 18 ): 7705 - 7710 .

WORDEN A Z , LEE J H , MOCK T , et al . Green evolution and dynamic adaptations revealed by genomes of the marine picoeukaryotes Micromonas [J ] . Science , 2009 , 324 ( 5924 ): 268 - 272 .

PROCHNIK S E , UMEN J , NEDELCU A M , et al . Genomic analysis of organismal complexity in the multicellular green alga Volvox carteri [J ] . Science , 2010 , 329 ( 5988 ): 223 - 226 .

BLANC-MATHIEU R , VERHELST B , DERELLE E , et al . An improved genome of the model marine alga Ostreococcus tauri unfolds by assessing Illumina de novo assemblies [J ] . BMC Genomics , 2014 , 15 : 1103 .

GAO C F , WANG Y , SHEN Y , et al . Oil accumulation mechanisms of the oleaginous microalga Chlorella protothecoides revealed through its genome, transcriptomes, and proteomes [J ] . BMC Genomics , 2014 , 15 : 582 .

BOWLER C , ALLEN A E , BADGER J H , et al . The Phaeodactylum genome reveals the evolutionary history of diatom genomes [J ] . Nature , 2008 , 456 ( 7219 ): 239 - 244 .

ARMBRUST E V , BERGES J A , BOWLER C , et al . The genome of the diatom Thalassiosira pseudonana : ecology, evolution, and metabolism [J ] . Science , 2004 , 306 ( 5693 ): 79 - 86 .

NOZAKI H , TAKANO H , MISUMI O , et al . A 100%-complete sequence reveals unusually simple genomic features in the hot-spring red alga Cyanidioschyzon merolae [J ] . BMC Biology , 2007 , 5 : 28 .

OHTA N , MATSUZAKI M , MISUMI O , et al . Complete sequence and analysis of the plastid genome of the unicellular red alga Cyanidioschyzon merolae [J ] . DNA Research , 2003 , 10 ( 2 ): 67 - 77 .

TAJIMA N , SATO S , MARUYAMA F , et al . Analysis of the complete plastid genome of the unicellular red alga Porphyridium purpureum [J ] . Journal of Plant Research , 2014 , 127 ( 3 ): 389 - 397 .

WANG D M , NING K , LI J , et al . Nannochloropsis genomes reveal evolution of microalgal oleaginous traits [J ] . PLoS Genetics , 2014 , 10 ( 1 ): e1004094 .

RADAKOVITS R , JINKERSON R E , FUERSTENBERG S I , et al . Draft genome sequence and genetic transformation of the oleaginous alga Nannochloropsis gaditana [J ] . Nature Communications , 2012 , 3 : 686 .

MUÑOZ C F , SÜDFELD C , NADUTHODI M I S , et al . Genetic engineering of microalgae for enhanced lipid production [J ] . Biotechnology Advances , 2021 , 52 : 107836 .

JIA J , HAN D X , GERKEN H G , et al . Molecular mechanisms for photosynthetic carbon partitioning into storage neutral lipids in Nannochloropsis oceanica under nitrogen-depletion conditions [J ] . Algal Research , 2015 , 7 : 66 - 77 .

LIU J , SUN Z , MAO X M , et al . Multiomics analysis reveals a distinct mechanism of oleaginousness in the emerging model alga Chromochloris zofingiensis [J ] . The Plant Journal , 2019 , 98 ( 6 ): 1060 - 1077 .

KAMALANATHAN M , PIERANGELINI M , SHEARMAN L A , et al . Impacts of nitrogen and phosphorus starvation on the physiology of Chlamydomonas reinhardtii [J ] . Journal of Applied Phycology , 2016 , 28 ( 3 ): 1509 - 1520 .

LIANG J B , IQBAL S , WEN F , et al . Phosphorus-induced lipid class alteration revealed by lipidomic and transcriptomic profiling in oleaginous microalga Nannochloropsis sp. PJ12 [J ] . Marine Drugs , 2019 , 17 ( 9 ): 519 .

JIN X J , GONG S Q , YANG B J , et al . Transcriptomic analysis for phosphorus limitation-induced β-glucans accumulation in Chlorella sorokiniana SCSIO 46784 during the early phase of growth [J ] . Algal Research , 2021 , 54 : 102208 .

HULATT C J , SMOLINA I , DOWLE A , et al . Proteomic and transcriptomic patterns during lipid remodeling in Nannochloropsis gaditana [J ] . International Journal of Molecular Sciences , 2020 , 21 ( 18 ): 6946 .

MANDOTRA S K , KUMAR P , SUSEELA M R , et al . Evaluation of fatty acid profile and biodiesel properties of microalga Scenedesmus abundans under the influence of phosphorus, pH and light intensities [J ] . Bioresource Technology , 2016 , 201 : 222 - 229 .

HE Q N , YANG H J , WU L , et al . Effect of light intensity on physiological changes, carbon allocation and neutral lipid accumulation in oleaginous microalgae [J ] . Bioresource Technology , 2015 , 191 : 219 - 228 .

LI Y L , SUN H , WANG Y N , et al . Integrated metabolic tools reveal carbon alternative in Isochrysis zhangjiangensis for fucoxanthin improvement [J ] . Bioresource Technology , 2022 , 347 : 126401 .

ATTA M , IDRIS A , BUKHARI A , et al . Intensity of blue LED light: a potential stimulus for biomass and lipid content in fresh water microalgae Chlorella vulgaris [J ] . Bioresource Technology , 2013 , 148 : 373 - 378 .

GAYTÁN-LUNA D E , OCHOA-ALFARO A E , ROCHA-URIBE A , et al . Effect of green and red light in lipid accumulation and transcriptional profile of genes implicated in lipid biosynthesis in Chlamydomonas reinhardtii [J ] . Biotechnology Progress , 2016 , 32 ( 6 ): 1404 - 1411 .

VIELER A , WU G X , TSAI C H , et al . Genome, functional gene annotation, and nuclear transformation of the heterokont oleaginous alga Nannochloropsis oceanica CCMP1779 [J ] . PLoS Genetics , 2012 , 8 ( 11 ): e1003064 .

POLINER E , BUSCH A W U , NEWTON L , et al . Aureochromes maintain polyunsaturated fatty acid content in Nannochloropsis oceanica [J ] . Plant Physiology , 2022 , 189 ( 2 ): 906 - 921 .

PATELOU M , INFANTE C , DARDELLE F , et al . Transcriptomic and metabolomic adaptation of Nannochloropsis gaditana grown under different light regimes [J ] . Algal Research , 2020 , 45 : 101735 .

WU L F , CHEN P C , LEE C M . The effects of nitrogen sources and temperature on cell growth and lipid accumulation of microalgae [J ] . International Biodeterioration & Biodegradation , 2013 , 85 : 506 - 510 .

CALHOUN S , BELL T A S , DAHLIN L R , et al . A multi-omic characterization of temperature stress in a halotolerant Scenedesmus strain for algal biotechnology [J ] . Communications Biology , 2021 , 4 : 333 .

XING G L , YUAN H L , YANG J S , et al . Integrated analyses of transcriptome, proteome and fatty acid profilings of the oleaginous microalga Auxenochlorella protothecoides UTEX 2341 reveal differential reprogramming of fatty acid metabolism in response to low and high temperatures [J ] . Algal Research , 2018 , 33 : 16 - 27 .

SU H , FENG J , LV J P , et al . Molecular mechanism of lipid accumulation and metabolism of oleaginous Chlorococcum sphacosum GD from soil under salt stress [J ] . International Journal of Molecular Sciences , 2021 , 22 ( 3 ): 1304 .

QU D H , MIAO X L . Carbon flow conversion induces alkali resistance and lipid accumulation under alkaline conditions based on transcriptome analysis in Chlorella sp. BLD [J ] . Chemosphere , 2021 , 265 : 129046 .

SINGH R D , SETHY S , GHOSH S , et al . UV and γ-radiation induced molecular changes for rapid lipid accumulation in Chlorella sorokiniana [J ] . Biomass and Bioenergy , 2022 , 163 : 106493 .

NAJAFABADI H A , MALEKZADEH M , JALILIAN F , et al . Effect of various carbon sources on biomass and lipid production of Chlorella vulgaris during nutrient sufficient and nitrogen starvation conditions [J ] . Bioresource Technology , 2015 , 180 : 311 - 317 .

UDAYAN A , ARUMUGAM M . Selective enrichment of eicosapentaenoic acid (20:5n-3) in N. oceanica CASA CC201 by natural auxin supplementation [J ] . Bioresource Technology , 2017 , 242 : 329 - 333 .

XUE L L , JIANG J G . Cultivation of Dunaliella tertiolecta intervened by triethylamine enhances the lipid content [J ] . Algal Research , 2017 , 25 : 136 - 141 .

ROESSLER P G . Changes in the activities of various lipid and carbohydrate biosynthetic enzymes in the diatom Cyclotella cryptica in response to silicon deficiency [J ] . Archives of Biochemistry and Biophysics , 1988 , 267 ( 2 ): 521 - 528 .

DUNAHAY T G , JARVIS E E , DAIS S S , et al . Manipulation of microalgal lipid production using genetic engineering [J ] . Applied Biochemistry and Biotechnology , 1996 , 57 : 223 - 231 .

GOMMA A E , LEE S K , SUN S M , et al . Improvement in oil production by increasing malonyl-CoA and glycerol-3-phosphate pools in Scenedesmus quadricauda [J ] . Indian Journal of Microbiology , 2015 , 55 ( 4 ): 447 - 455 .

CHEN J W , LIU W J , HU D X , et al . Identification of a malonyl CoA-acyl carrier protein transacylase and its regulatory role in fatty acid biosynthesis in oleaginous microalga Nannochloropsis oceanica [J ] . Biotechnology and Applied Biochemistry , 2017 , 64 ( 5 ): 620 - 626 .

RENGEL R , SMITH R T , HASLAM R P , et al . Overexpression of acetyl-CoA synthetase (ACS) enhances the biosynthesis of neutral lipids and starch in the green microalga Chlamydomonas reinhardtii [J ] . Algal Research , 2018 , 31 : 183 - 193 .

YAN J F , CHENG R B , LIN X Z , et al . Overexpression of acetyl-CoA synthetase increased the biomass and fatty acid proportion in microalga Schizochytrium [J ] . Applied Microbiology and Biotechnology , 2013 , 97 ( 5 ): 1933 - 1939 .

OSADA K , MAEDA Y , YOSHINO T , et al . Enhanced NADPH production in the pentose phosphate pathway accelerates lipid accumulation in the oleaginous diatom Fistulifera solaris [J ] . Algal Research , 2017 , 23 : 126 - 134 .

NIU Y F , WANG X , HU D X , et al . Molecular characterization of a glycerol-3-phosphate acyltransferase reveals key features essential for triacylglycerol production in Phaeodactylum tricornutum [J ] . Biotechnology for Biofuels , 2016 , 9 : 60 .

MUÑOZ C F , WEUSTHUIS R A , D'ADAMO S , et al . Effect of single and combined expression of lysophosphatidic acid acyltransferase, glycerol-3-phosphate acyltransferase, and diacylglycerol acyltransferase on lipid accumulation and composition in Neochloris oleoabundans [J ] . Frontiers in Plant Science , 2019 , 10 : 1573 .

MARAVI D K , KUMAR S , SHARMA P K , et al . Ectopic expression of AtDGAT1 , encoding diacylglycerol O -acyltransferase exclusively committed to TAG biosynthesis, enhances oil accumulation in seeds and leaves of Jatropha [J ] . Biotechnology for Biofuels , 2016 , 9 : 226 .

WANG Z K , HUANG W J , CHANG J M , et al . Overexpression of SiDGAT1 , a gene encoding acyl-CoA: diacylglycerol acyltransferase from Sesamum indicum L. increases oil content in transgenic Arabidopsis and soybean [J ] . Plant Cell, Tissue and Organ Culture (PCTOC) , 2014 , 119 ( 2 ): 399 - 410 .

NIU Y F , ZHANG M H , LI D W , et al . Improvement of neutral lipid and polyunsaturated fatty acid biosynthesis by overexpressing a type 2 diacylglycerol acyltransferase in marine diatom Phaeodactylum tricornutum [J ] . Marine Drugs , 2013 , 11 ( 11 ): 4558 - 4569 .

LI D W , CEN S Y , LIU Y H , et al . A type 2 diacylglycerol acyltransferase accelerates the triacylglycerol biosynthesis in heterokont oleaginous microalga Nannochloropsis oceanica [J ] . Journal of Biotechnology , 2016 , 229 : 65 - 71 .

DENG X D , GU B , LI Y J , et al . The roles of acyl-CoA: diacylglycerol acyltransferase 2 genes in the biosynthesis of triacylglycerols by the green algae Chlamydomonas reinhardtii [J ] . Molecular Plant , 2012 , 5 ( 4 ): 945 - 947 .

COUSO I , PÉREZ-PÉREZ M E , FORD M M , et al . Phosphorus availability regulates TORC1 signaling via LST8 in Chlamydomonas [J ] . The Plant Cell , 2020 , 32 ( 1 ): 69 - 80 .

NGAN C Y , WONG C H , CHOI C , et al . Lineage-specific chromatin signatures reveal a regulator of lipid metabolism in microalgae [J ] . Nature Plants , 2015 , 1 : 15107 .

BAJHAIYA A K , DEAN A P , ZEEF L A H , et al . PSR1 is a global transcriptional regulator of phosphorus deficiency responses and carbon storage metabolism in Chlamydomonas reinhardtii [J ] . Plant Physiology , 2016 , 170 ( 3 ): 1216 - 1234 .

JIA B , XIE X F , WU M , et al . Understanding the functions of endogenous DOF transcript factor in Chlamydomonas reinhardtii [J ] . Biotechnology for Biofuels , 2019 , 12 : 67 .

ZHANG J H , HAO Q , BAI L L , et al . Overexpression of the soybean transcription factor GmDof4 significantly enhances the lipid content of Chlorella ellipsoidea [J ] . Biotechnology for Biofuels , 2014 , 7 : 128 .

LI D W , BALAMURUGAN S , YANG Y F , et al . Transcriptional regulation of microalgae for concurrent lipid overproduction and secretion [J ] . Science Advances , 2019 , 5 ( 1 ): eaau3795 .

YAMAOKA Y , SHIN S , CHOI B Y , et al . The bZIP1 transcription factor regulates lipid remodeling and contributes to ER stress management in Chlamydomonas reinhardtii [J ] . The Plant Cell , 2019 , 31 ( 5 ): 1127 - 1140 .

AJJAWI I , VERRUTO J , AQUI M , et al . Lipid production in Nannochloropsis gaditana is doubled by decreasing expression of a single transcriptional regulator [J ] . Nature Biotechnology , 2017 , 35 ( 7 ): 647 - 652 .

RADAKOVITS R , EDUAFO P M , POSEWITZ M C . Genetic engineering of fatty acid chain length in Phaeodactylum tricornutum [J ] . Metabolic Engineering , 2011 , 13 : 89 - 95 .

GONG Y M , GUO X J , WAN X , et al . Characterization of a novel thioesterase (PtTE) from Phaeodactylum tricornutum [J ] . Journal of Basic Microbiology , 2011 , 51 ( 6 ): 666 - 672 .

PENG K T , ZHENG C N , XUE J , et al . Delta 5 fatty acid desaturase upregulates the synthesis of polyunsaturated fatty acids in the marine diatom Phaeodactylum tricornutum [J ] . Journal of Agricultural and Food Chemistry , 2014 , 62 ( 35 ): 8773 - 8776 .

HAMILTON M L , HASLAM R P , NAPIER J A , et al . Metabolic engineering of Phaeodactylum tricornutum for the enhanced accumulation of omega-3 long chain polyunsaturated fatty acids [J ] . Metabolic Engineering , 2014 , 22 : 3 - 9 .

SHI H S , LUO X , WU R N , et al . Production of eicosapentaenoic acid by application of a delta-6 desaturase with the highest ALA catalytic activity in algae [J ] . Microbial Cell Factories , 2018 , 17 : 7 .

POLINER E , PULMAN J A , ZIENKIEWICZ K , et al . 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 [J ] . Plant Biotechnology Journal , 2018 , 16 : 298 - 309 .

NORASHIKIN M N , LOH S H , AZIZ A , et al . Metabolic engineering of fatty acid biosynthesis in Chlorella vulgaris using an endogenous omega-3 fatty acid desaturase gene with its promoter [J ] . Algal Research , 2018 , 31 : 262 - 275 .

WANG X , DONG H P , WEI W , et al . Dual expression of plastidial GPAT1 and LPAT1 regulates triacylglycerol production and the fatty acid profile in Phaeodactylum tricornutum [J ] . Biotechnology for Biofuels , 2018 , 11 : 318 .

LIU J , LIU M J , PAN Y F , et al . Metabolic engineering of the oleaginous alga Nannochloropsis for enriching eicosapentaenoic acid in triacylglycerol by combined pulling and pushing strategies [J ] . Metabolic Engineering , 2022 , 69 : 163 - 174 .

XIN Y , LU Y D , LEE Y Y , et al . Producing designer oils in industrial microalgae by rational modulation of Co-evolving type-2 diacylglycerol acyltransferases [J ] . Molecular Plant , 2017 , 10 ( 12 ): 1523 - 1539 .

LU N , WEI D , JIANG X L , et al . Regulation of lipid metabolism in the snow alga Chlamydomonas nivalis in response to NaCl stress: an integrated analysis by cytomic and lipidomic approaches [J ] . Process Biochemistry , 2012 , 47 ( 7 ): 1163 - 1170 .

SUN H , REN Y Y , MAO X M , et al . Harnessing C/N balance of Chromochloris zofingiensis to overcome the potential conflict in microalgal production [J ] . Communications Biology , 2020 , 3 : 186 .

SUN H , WU T , CHEN S H Y , et al . Powerful tools for productivity improvements in microalgal production [J ] . Renewable and Sustainable Energy Reviews , 2021 , 152 : 111609 .

KONG F T , LIANG Y X , LÉGERET B , et al . Chlamydomonas carries out fatty acid β-oxidation in ancestral peroxisomes using a bona fide acyl-CoA oxidase [J ] . The Plant Journal , 2017 , 90 ( 2 ): 358 - 371 .

CRAIG W , GARGANO D , SCOTTI N , et al . Direct gene transfer in potato: a comparison of particle bombardment of leaf explants and PEG-mediated transformation of protoplasts [J ] . Plant Cell Reports , 2005 , 24 ( 10 ): 603 - 611 .

NG S L , HARIKRISHNA J A , ABU BAKAR F , et al . Heterologous expression of the Streptococcus pneumoniae yoeB and pezT toxin genes is lethal in Chlorella vulgaris [J ] . Algal Research , 2016 , 19 : 21 - 29 .

YANG B , LIU J , LIU B , et al . Development of a stable genetic system for Chlorella vulgaris —a promising green alga for CO 2 biomitigation [J ] . Algal Research , 2015 , 12 : 134 - 141 .

TALEBI A F , TOHIDFAR M , TABATABAEI M , et al . Genetic manipulation, a feasible tool to enhance unique characteristic of Chlorella vulgaris as a feedstock for biodiesel production [J ] . Molecular Biology Reports , 2013 , 40 ( 7 ): 4421 - 4428 .

LIU L L , WANG Y Q , ZHANG Y C , et al . Development of a new method for genetic transformation of the green alga Chlorella ellipsoidea [J ] . Molecular Biotechnology , 2013 , 54 ( 2 ): 211 - 219 .

BAI L L , YIN W B , CHEN Y H , et al . A new strategy to produce a defensin: stable production of mutated NP-1 in nitrate reductase-deficient Chlorella ellipsoidea [J ] . PLoS One , 2013 , 8 ( 1 ): e54966 .

HUANG C C , CHEN M W , HSIEH J L , et al . Expression of mercuric reductase from Bacillus megaterium MB1 in eukaryotic microalga Chlorella sp. DT: an approach for mercury phytoremediation [J ] . Applied Microbiology and Biotechnology , 2006 , 72 ( 1 ): 197 - 205 .

LIU J , SUN Z , GERKEN H , et al . Genetic engineering of the green alga Chlorella zofingiensis : a modified norflurazon-resistant phytoene desaturase gene as a dominant selectable marker [J ] . Applied Microbiology and Biotechnology , 2014 , 98 ( 11 ): 5069 - 5079 .

LI F J , GAO D W , HU H H . High-efficiency nuclear transformation of the oleaginous marine Nannochloropsis species using PCR product [J ] . Bioscience, Biotechnology, and Biochemistry , 2014 , 78 ( 5 ): 812 - 817 .

KILIAN O , BENEMANN C S E , NIYOGI K K , et al . High-efficiency homologous recombination in the oil-producing alga Nannochloropsis sp .[J ] . Proceedings of the National Academy of Sciences of the United States of America , 2011 , 108 ( 52 ): 21265 - 21269 .

KANG N K , JEON S , KWON S , et al . Effects of overexpression of a bHLH transcription factor on biomass and lipid production in Nannochloropsis salina [J ] . Biotechnology for Biofuels , 2015 , 8 : 200 .

WEI L , XIN Y , WANG Q T , et al . RNAi-based targeted gene knockdown in the model oleaginous microalgae Nannochloropsis oceanica [J ] . The Plant Journal , 2017 , 89 ( 6 ): 1236 - 1250 .

SHIH C H , CHEN H Y , LEE H C , et al . Purple chromoprotein gene serves as a new selection marker for transgenesis of the microalga Nannochloropsis oculata [J ] . PLoS One , 2015 , 10 ( 3 ): e0120780 .

SIMON D P , ANILA N , GAYATHRI K , et al . Heterologous expression of β-carotene hydroxylase in Dunaliella salina by Agrobacterium -mediated genetic transformation [J ] . Algal Research , 2016 , 18 : 257 - 265 .

FENG S Y , FENG W P , ZHAO L , et al . Preparation of transgenic Dunaliella salina for immunization against white spot syndrome virus in crayfish [J ] . Archives of Virology , 2014 , 159 ( 3 ): 519 - 525 .

姜国忠 , 吕玉民 , 牛向丽 , 等 . 盐藻肌动蛋白基因启动子驱动的 bar 基因表达作为核转化筛选标记 [J ] . 遗传学报 , 2005 , 32 ( 4 ): 424 - 433 .

JIANG G Z , LÜ Y M , NIU X L , et al . The actin gene promoter-driven bar as a dominant selectable marker for nuclear transformation of Dunaliella salina [J ] . Acta Genetica Sinica , 2005 , 32 ( 4 ): 424 - 433 .

ZHANG J Y , JEON H , SIM S J , et al . Homologous sense and antisense expression of a gene in Dunaliella tertiolecta [J ] . Planta , 2015 , 242 ( 4 ): 1051 - 1058 .

GEORGIANNA D R , HANNON M J , MARCUSCHI M , et al . Production of recombinant enzymes in the marine alga Dunaliella tertiolecta [J ] . Algal Research , 2013 , 2 ( 1 ): 2 - 9 .

KADONO T , MIYAGAWA-YAMAGUCHI A , KIRA N , et al . Characterization of marine diatom-infecting virus promoters in the model diatom Phaeodactylum tricornutum [J ] . Scientific Reports , 2015 , 5 : 18708 .

ZHANG C Y , HU H H . High-efficiency nuclear transformation of the diatom Phaeodactylum tricornutum by electroporation [J ] . Marine Genomics , 2014 , 16 : 63 - 66 .

POULSEN N , CHESLEY P M , KRÖGER N . Molecular genetic manipulation of the diatom Thalassiosira pseudonana (Bacillariophyceae) [J ] . Journal of Phycology , 2006 , 42 ( 5 ): 1059 - 1065 .

SHARON-GOJMAN R , MAIMON E , LEU S , et al . Advanced methods for genetic engineering of Haematococcus pluvialis (Chlorophyceae, Volvocales) [J ] . Algal Research , 2015 , 10 : 8 - 15 .

GUTIÉRREZ C L , GIMPEL J , ESCOBAR C , et al . Chloroplast genetic tool for the green microalgae Haematococcus pluvialis (Chlorophyceae, Volvocales) 1 [J ] . Journal of Phycology , 2012 , 48 ( 4 ): 976 - 983 .

STEINBRENNER J , SANDMANN G . Transformation of the green alga Haematococcus pluvialis with a phytoene desaturase for accelerated astaxanthin biosynthesis [J ] . Applied and Environmental Microbiology , 2006 , 72 ( 12 ): 7477 - 7484 .

KATHIRESAN S , CHANDRASHEKAR A , RAVISHANKAR G A , et al . Agrobacterium -mediated transformation in the green alga Haematococcus pluvialis (Chlorophyceae, Volvocales) [J ] . Journal of Phycology , 2009 , 45 ( 3 ): 642 - 649 .

MUTO M , FUKUDA Y , NEMOTO M , et al . Establishment of a genetic transformation system for the marine pennate diatom Fistulifera sp. strain JPCC DA0580—a high triglyceride producer [J ] . Marine Biotechnology , 2013 , 15 ( 1 ): 48 - 55 .

JAEGER D , HÜBNER W , HUSER T , et al . Nuclear transformation and functional gene expression in the oleaginous microalga Monoraphidium neglectum [J ] . Journal of Biotechnology , 2017 , 249 : 10 - 15 .

CHUNGJATUPORNCHAI W , KITRAKSA P , FA-AROONSAWAT S . Stable nuclear transformation of the oleaginous microalga Neochloris oleoabundans by electroporation [J ] . Journal of Applied Phycology , 2016 , 28 ( 1 ): 191 - 199 .

BERRIOS H , ZAPATA M , RIVAS M . A method for genetic transformation of Botryococcus braunii using a cellulase pretreatment [J ] . Journal of Applied Phycology , 2016 , 28 ( 1 ): 201 - 208 .

ÚBEDA-MÍNGUEZ P , CHILEH T , DAUTOR Y , et al . Tools for microalgal biotechnology: development of an optimized transformation method for an industrially promising microalga— Tetraselmis chuii [J ] . Journal of Applied Phycology , 2015 , 27 ( 1 ): 223 - 232 .

ORTIZ-MATAMOROS M F , VILLANUEVA M A , ISLAS-FLORES T . Transient transformation of cultured photosynthetic dinoflagellates ( Symbiodinium spp.) with plant-targeted vectors [J ] . Ciencias Marinas , 2015 , 41 ( 1 ): 21 - 32 .

ZORIN B , GRUNDMAN O , KHOZIN-GOLDBERG I , et al . 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 [J ] . PLoS One , 2014 , 9 ( 8 ): e105223 .

PRASAD B , VADAKEDATH N , JEONG H J , et al . Agrobacterium tumefaciens -mediated genetic transformation of haptophytes ( Isochrysis species) [J ] . Applied Microbiology and Biotechnology , 2014 , 98 ( 20 ): 8629 - 8639 .

RATHOD J P , PRAKASH G , PANDIT R , et al . Agrobacterium-mediated transformation of promising oil-bearing marine algae Parachlorella kessleri [J ] . Photosynthesis Research , 2013 , 118 ( 1/2 ): 141 - 146 .

HONG W K , HEO S Y , OH B R , et al . A transgene expression system for the marine microalgae Aurantiochytrium sp. KRS101 using a mutant allele of the gene encoding ribosomal protein L44 as a selectable transformation marker for cycloheximide resistance [J ] . Bioprocess and Biosystems Engineering , 2013 , 36 ( 9 ): 1191 - 1197 .

GUO S L , ZHAO X Q , TANG Y , et al . Establishment of an efficient genetic transformation system in Scenedesmus obliquus [J ] . Journal of Biotechnology , 2013 , 163 ( 1 ): 61 - 68 .

CHENG R B , MA R J , LI K , et al . Agrobacterium tumefaciens mediated transformation of marine microalgae Schizochytrium [J ] . Microbiological Research , 2012 , 167 ( 3 ): 179 - 186 .

CUI Y L , WANG J F , JIANG P , et al . Transformation of Platymonas ( Tetraselmis ) subcordiformis (Prasinophyceae, Chlorophyta) by agitation with glass beads [J ] . World Journal of Microbiology and Biotechnology , 2010 , 26 ( 9 ): 1653 - 1657 .

LAPIDOT M , RAVEH D , SIVAN A , et al . Stable chloroplast transformation of the unicellular red alga Porphyridium species [J ] . Plant Physiology , 2002 , 129 : 7 - 12 .

KAWATA Y , YANO S , KOJIMA H , et al . Transformation of Spirulina platensis strain C1 ( Arthrospira sp. PCC9438) with Tn5 transposase -transposon DNA-cation liposome complex [J ] . Marine Biotechnology , 2004 , 6 ( 4 ): 355 - 363 .

JAENICKE L , KUHNE W , SPESSERT R , et al . Cell-wall lytic enzymes (autolysins) of Chlamydomonas reinhardtii are (hydroxy)proline-specific proteases [J ] . European Journal of Biochemistry , 1987 , 170 ( 1/2 ): 485 - 491 .

ZHU Z , SUN J , FA Y , et al . Enhancing microalgal lipid accumulation for biofuel production [J ] . Frontiers in Microbiology , 2022 , 13 : 1024441 .

WU X G , GIRIDHAR BABU A , KIM B L , et al . Ultrasound-mediated intracellular delivery of fluorescent dyes and DNA into microalgal cells [J ] . Algal Research , 2016 , 15 : 210 - 216 .

KINDLE K L , SCHNELL R A , FERNÁNDEZ E , et al . Stable nuclear transformation of Chlamydomonas using the Chlamydomonas gene for nitrate reductase [J ] . The Journal of Cell Biology , 1989 , 109 ( 6 ): 2589 - 2601 .

SØRENSEN I , FEI Z J , ANDREAS A , et al . Stable transformation and reverse genetic analysis of Penium margaritaceum : a platform for studies of charophyte green algae, the immediate ancestors of land plants [J ] . The Plant Journal , 2014 , 77 ( 3 ): 339 - 351 .

JIA Y L , XUE L X , LIU H T , et al . Characterization of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene from the halotolerant alga Dunaliella salina and inhibition of its expression by RNAi [J ] . Current Microbiology , 2009 , 58 ( 5 ): 426 - 431 .

RANDHAWA S , SENGAR S . The evolution and history of gene editing technologies [M/OL ] // Progress in molecular biology and translational science . Amsterdam : Elsevier , 2021 , 178 : 1 - 62 [2023-07-01] . https://www.sciencedirect.com/science/article/abs/pii/S1877117321000028?via%3Dihub https://www.sciencedirect.com/science/article/abs/pii/S1877117321000028?via%3Dihub .

SIZOVA I , GREINER A , AWASTHI M , et al . Nuclear gene targeting in Chlamydomonas using engineered zinc-finger nucleases [J ] . The Plant Journal , 2013 , 73 ( 5 ): 873 - 882 .

DABOUSSI F , LEDUC S , MARÉCHAL A , et al . Genome engineering empowers the diatom Phaeodactylum tricornutum for biotechnology [J ] . Nature Communications , 2014 , 5 : 3831 .

WEYMAN P D , BEERI K , LEFEBVRE S C , et al . Inactivation of Phaeodactylum tricornutum urease gene using transcription activator-like effector nuclease-based targeted mutagenesis [J ] . Plant Biotechnology Journal , 2015 , 13 ( 4 ): 460 - 470 .

NGUYEN T H T , PARK S , JEONG J , et al . Enhancing lipid productivity by modulating lipid catabolism using the CRISPR-Cas9 system in Chlamydomonas [J ] . Journal of Applied Phycology , 2020 , 32 ( 5 ): 2829 - 2840 .

FENG S Y , HU L N , ZHANG Q H , et al . CRISPR/Cas technology promotes the various application of Dunaliella salina system [J ] . Applied Microbiology and Biotechnology , 2020 , 104 ( 20 ): 8621 - 8630 .

LIN W R , NG I S . Development of CRISPR/Cas9 system in Chlorella vulgaris FSP-E to enhance lipid accumulation [J ] . Enzyme and Microbial Technology , 2020 , 133 : 109458 .

VERRUTO J , FRANCIS K , WANG Y J , et al . Unrestrained markerless trait stacking in Nannochloropsis gaditana through combined genome editing and marker recycling technologies [J ] . Proceedings of the National Academy of Sciences of the United States of America , 2018 , 115 ( 30 ): E7015 - E7022 .

WANG Q T , GONG Y H , HE Y H , et al . Genome engineering of Nannochloropsis with hundred-kilobase fragment deletions by Cas9 cleavages [J ] . The Plant Journal , 2021 , 106 ( 4 ): 1148 - 1162 .

BAEK K , KIM D H , JEONG J , et al . DNA-free two-gene knockout in Chlamydomonas reinhardtii via CRISPR-Cas9 ribonucleoproteins [J ] . Scientific Reports , 2016 , 6 : 30620 .

SCHRODA M , BLÖCKER D , BECK C F . The HSP70A promoter as a tool for the improved expression of transgenes in Chlamydomonas [J ] . The Plant Journal , 2000 , 21 ( 2 ): 121 - 131 .

FRANKLIN S , NGO B , EFUET E , et al . Development of a GFP reporter gene for Chlamydomonas reinhardtii chloroplast [J ] . The Plant Journal , 2002 , 30 ( 6 ): 733 - 744 .

JIA Y L , LI S K , ALLEN G , et al . A novel glyceraldehyde-3-phosphate dehydrogenase (GAPDH) promoter for expressing transgenes in the halotolerant alga Dunaliella salina [J ] . Current Microbiology , 2012 , 64 ( 5 ): 506 - 513 .

HOU Q T , QIU S , LIU Q , et al . Selenoprotein-transgenic Chlamydomonas reinhardtii [J ] . Nutrients , 2013 , 5 ( 3 ): 624 - 636 .

CROZET P , NAVARRO F J , WILLMUND F , et al . Birth of a photosynthetic chassis: a MoClo toolkit enabling synthetic biology in the microalga Chlamydomonas reinhardtii [J ] . ACS Synthetic Biology , 2018 , 7 ( 9 ): 2074 - 2086 .

SUN H , REN Y Y , FAN Y W , et al . Systematic metabolic tools reveal underlying mechanism of product biosynthesis in Chromochloris zofingiensis [J ] . Bioresource Technology , 2021 , 337 : 125406 .

SUN H , WANG Y X , HE Y J , et al . Microalgae-derived pigments for the food industry [J ] . Marine Drugs , 2023 , 21 ( 2 ): 82 .

SCHUHMANN H , LIM D K , SCHENK P M . Perspectives on metabolic engineering for increased lipid contents in microalgae [J ] . Biofuels , 2012 , 3 ( 1 ): 71 - 86 .

YAMORI W , SHIKANAI T . Physiological functions of cyclic electron transport around photosystemⅠin sustaining photosynthesis and plant growth [J ] . Annual Review of Plant Biology , 2016 , 67 : 81 - 106 .

XUE J , BALAMURUGAN S , LI D W , et al . Glucose-6-phosphate dehydrogenase as a target for highly efficient fatty acid biosynthesis in microalgae by enhancing NADPH supply [J ] . Metabolic Engineering , 2017 , 41 : 212 - 221 .

SCHIEVANO A , SCIARRIA T P , VANBROEKHOVEN K , et al . Electro-fermentation-merging electrochemistry with fermentation in industrial applications [J ] . Trends in Biotechnology , 2016 , 34 ( 11 ): 866 - 878 .

CHOI O , KIM T , WOO H M , et al . Electricity-driven metabolic shift through direct electron uptake by electroactive heterotroph Clostridium pasteurianum [J ] . Scientific Reports , 2014 , 4 : 6961 .

MICHELET L , ZAFFAGNINI M , MORISSE S , et al . Redox regulation of the Calvin-Benson cycle: something old, something new [J ] . Frontiers in Plant Science , 2013 , 4 : 470 .

KUREK I , CHANG T K , BERTAIN S M , et al . Enhanced thermostability of Arabidopsis Rubisco activase improves photosynthesis and growth rates under moderate heat stress [J ] . The Plant Cell , 2007 , 19 ( 10 ): 3230 - 3241 .

YAMORI W , MASUMOTO C , FUKAYAMA H , et al . Rubisco activase is a key regulator of non-steady-state photosynthesis at any leaf temperature and, to a lesser extent, of steady-state photosynthesis at high temperature [J ] . The Plant Journal , 2012 , 71 ( 6 ): 871 - 880 .

VUORISTO K S , MARS A E , SANDERS J P M , et al . Metabolic engineering of TCA cycle for production of chemicals [J ] . Trends in Biotechnology , 2016 , 34 ( 3 ): 191 - 197 .

SAINI R , KAPOOR R , KUMAR R , et al . CO 2 utilizing microbes—a comprehensive review [J ] . Biotechnology Advances , 2011 , 29 ( 6 ): 949 - 960 .

XIONG W , MORGAN J A , UNGERER J , et al . The plasticity of cyanobacterial metabolism supports direct CO 2 conversion to ethylene [J ] . Nature Plants , 2015 , 1 : 15053 .

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