Overexpression of S-Adenosylmethionine Synthetase in Recombinant Chlamydomonas for Enhanced Lipid Production
1
조회
0
다운로드
1
인용
0
참고문헌
키워드
초록
Microalgae are attracting much attention as promising, eco-friendly producers of bioenergy due to their fast growth, absorption of carbon dioxide from the atmosphere, and production capacity in wastewater and salt water. However, microalgae can only accumulate large quantities of lipid in abiotic stress, which reduces productivity by decreasing cell growth. In this study, the strategy was investigated to increase cell viability and lipid production by overexpressing S-adenosylmethionine (SAM) synthetase (SAMS) in the microalga Chlamydomonas reinhardtii. SAM is a substance that plays an important role in various intracellular biochemical reactions, such as cell proliferation and stress response, and the overexpression of SAMS could allow cells to withstand the abiotic stress and increase productivity. Compared to wild-type C. reinhardtii, recombinant cells overexpressing SAMS grew 1.56-fold faster and produced 1.51-fold more lipids in a nitrogen-depleted medium. Furthermore, under saline-stress conditions, the survival rate and lipid accumulation were 1.56 and 2.04 times higher in the SAMS-overexpressing strain, respectively. These results suggest that the overexpression of SAMS in recombinant C. reinhardtii has high potential in the industrial-scale production of biofuels and various other high-value-added materials.
Abstract
Microalgae are attracting much attention as promising, eco-friendly producers of bioenergy due to their fast growth, absorption of carbon dioxide from the atmosphere, and production capacity in wastewater and salt water. However, microalgae can only accumulate large quantities of lipid in abiotic stress, which reduces productivity by decreasing cell growth. In this study, the strategy was investigated to increase cell viability and lipid production by overexpressing S-adenosylmethionine (SAM) synthetase (SAMS) in the microalga Chlamydomonas reinhardtii. SAM is a substance that plays an important role in various intracellular biochemical reactions, such as cell proliferation and stress response, and the overexpression of SAMS could allow cells to withstand the abiotic stress and increase productivity. Compared to wild-type C. reinhardtii, recombinant cells overexpressing SAMS grew 1.56-fold faster and produced 1.51-fold more lipids in a nitrogen-depleted medium. Furthermore, under saline-stress conditions, the survival rate and lipid accumulation were 1.56 and 2.04 times higher in the SAMS-overexpressing strain, respectively. These results suggest that the overexpression of SAMS in recombinant C. reinhardtii has high potential in the industrial-scale production of biofuels and various other high-value-added materials.
참고문헌 (37)
- Oh H, "South Korea? 2050 Carbon neutrality policy", East Asian Policy, 13, pp.33~46 (2021)
- Rodolfi L, "Microalgae for oil: strain selection, induction of lipid synthesis and outdoor mass cultivation in a low‐cost photobioreactor", Biotechnol.Bbioeng, 102, pp.100~112 (2009)
- 김종태, "미세조류 배양을 위한 광생물반응기 개발의 기술동향", 한국정밀공학회지, 28(2), pp.125~132 (2011), 한국정밀공학회
- Work VH, "Increased lipid accumulation in the Chlamydomonas reinhardtii sta7-10 starchless isoamylase mutant and increased carbohydrate synthesis in complemented strains", Eukaryot. Cell, 9, pp.1251~1261 (2010)
- Paliwal C, "Abiotic stresses as tools for metabolites in microalgae", Bioresour. Technol, 244, pp.1216~1226 (2017)
- Colina F, "Genome-wide identification and characterization of CKIN", SnRK gene family in Chlamydomonas reinhardtii, 2019, pp.9, 350~
- Sasso S, "The natural history of model organisms: From molecular manipulation of domesticated Chlamydomonas reinhardtii to survival in nature", Elife, 7, pp.e39233~ (2018)
- Mayfield SP, "Expression of human antibodies in eukaryotic micro-algae", Vaccine, 23, pp.1828~1832 (2005)
- Park JJ, "The response of Chlamydomonas reinhardtii to nitrogen deprivation: a systems biology analysis", Plant J, 81, pp.611~624 (2015)
- Baek K, "DNA-free two-gene knockout in Chlamydomonas reinhardtii via CRISPR-Cas9 ribonucleoproteins", Sci. Rep, 6, pp.30620~ (2016)
- Lin Y-H, "Overexpression of ferredoxin, PETF, enhances tolerance to heat stress in Chlamydomonas reinhardtii", Int. J. Mol. Sci, 14, pp.20913~20929 (2013)
- Kim J, "Site-specific gene knock-out and on-site heterologous gene overexpression in Chlamydomonas reinhardtii via a CRISPR-Cas9-mediated knock-in method", Front. Plant Sci, 11, pp.306~ (2020)
- Park Seo-jeong, "Improved tolerance of recombinant Chlamydomonas rainhardtii with putative 2-amino-3-carboxymuconate-6-semialdehyde decarboxylase from Pyropia yezoensis to nitrogen starvation", The Journal of Microbiology, 60(1), pp.63~69 (2022), 한국미생물학회[DOI]
- Lu SC, "S-adenosylmethionine", Int. J. Biochem. Cell Biol, 32, pp.391~395 (2000)
- Yoon S, "Structural and functional characterization of S-adenosylmethionine (SAM) synthetase from Pichia ciferrii", Bioproc. Biosyst. Eng, 35, pp.173~181 (2012)
- Hayashi T, "S-adenosylmethionine synthetase is required for cell growth, maintenance of G0 phase, and termination of quiescence in fission yeast", Iscience, 5, pp.38~51 (2018)
- Ma C, "Overexpression of S-adenosyl-L-methionine synthetase 2 from sugar beet M14increased Arabidopsis tolerance to salt and oxidative stress", Int. J. Mol. Sci, 18, pp.847~ (2017)
- 한종원, "Characterization of S-adenosylmethionine Synthetase Gene from Red Algae, Pyropia tenera", Plant Breeding and Biotechnology, 4(4), pp.475~483 (2016), 한국육종학회[DOI]
- Roy M, "Overexpression of S-adenosylmethionine decarboxylase gene in rice increases polyamine level and enhances sodium chloride-stress tolerance", Plant Sci, 163, pp.987~992 (2002)
- Hazarika P, "Biotic and abiotic stress tolerance in transgenic tomatoes by constitutive expression of Sadenosylmethionine decarboxylase gene", Physiol. Mol. Biol. Plants, 17, pp.115~128 (2011)
- Merchant SS, "The Chlamydomonas genome reveals the evolution of key animal and plant functions", Science, 318, pp.245~250 (2007)
- Kim E, "IDENTIFICATION OF THE HIGH‐TEMPERATURE RESPONSE GENES FROM PORPHYRA SERIATA (RHODOPHYTA) EXPRESSION SEQUENCE TAGS AND ENHANCEMENT OF HEAT TOLERANCE OF CHLAMYDOMONAS (CHLOROPHYTA) BY EXPRESSION OF THE PORPHYRA HTR2 GENE 1", J. Phycol, 47, pp.821~828 (2011)
- Livak KJ, "Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method", Mthods, 25, pp.402~408 (2001)
- Kim D-J, "Accumulation of S-adenosyl-L-methionine enhances production of actinorhodin but inhibits sporulation in Streptomyces lividans TK23", J. Bacteriol, 185, pp.592~600 (2003)
- Birsan C, "A novel HPLC-PDA-MS method for S-adenosylmethionine and S-adenosylhomocysteine routine analysis", Anal. Lett, 43, pp.793~803 (2010)
- Cirulis JT, "Optimization of staining conditions for microalgae with three lipophilic dyes to reduce precipitation and fluorescence variability", Cytometry A, 81, pp.618~626 (2012)
- Sartory D, "Extraction of chlorophyll a from freshwater phytoplankton for spectrophotometric analysis", Hydrobiologia, 114, pp.177~187 (1984)
- Greenbaum D, "Comparing protein abundance and mRNA expression levels on a genomic scale", Genome Biol, 4, pp.117~ (2003)
- Lin HY, "Polyamines in microalgae: Something borrowed, something new", Mar. Drugs, 17, pp.1~ (2018)
- Eloranta TO, "S-adenosylmethionine metabolism and its relation to polyamine synthesis in rat liver. Effect of nutritional state, adrenal function, some drugs and partial hepatectomy", Biochem. J, 168, pp.179~185 (1977)
- Gevrekci AÖ, "The roles of polyamines in microorganisms", World J. Microbiol. Biotechnol, 33, pp.204~ (2017)
- Rezayian M, "Oxidative damage and antioxidative system in algae", Toxicol. Rrep, 6, pp.1309~1313 (2019)
- Sachdev S, "Abiotic stress and reactive oxygen species: generation, signaling, and defense mechanisms", Antioxidants (Basel), 10, pp.277~ (2021)
- Liu JH, "Polyamines function in stress tolerance: from synthesis to regulation", Front. Plant Sci, 6, pp.827~ (2015)
- Li-Beisson Y, "Metabolism of acyl‐lipids in Chlamydomonas reinhardtii", Plant J, 82, pp.504~522 (2015)
- Wase N, "Integrated quantitative analysis of nitrogen stress response in Chlamydomonas reinhardtii using metabolite and protein profiling", J. Proteome Res, 13, pp.1373~1396 (2014)
- Lee B-S, "Overexpression of fructose-1, 6-bisphosphate aldolase 1 enhances accumulation of fatty acids in Chlamydomonas reinhardtii", Algal Res, 47, pp.101825~ (2020)
