Supercritical carbon dioxide extraction of astaxanthin from Corynebacterium glutamicum

Abstract

Astaxanthin, a red carotenoid with potent antioxidant properties, holds significant value in the feed, cosmetics, and nutraceutical industries. While traditionally sourced from microalgae, Corynebacterium glutamicum, a well-established industrial microorganism, has been engineered to serve as an efficient host for astaxanthin production. As astaxanthin integrates into the cellular membrane, effective extraction methods are essential to access this valuable compound. In this study, a sustainable batch extraction process using supercritical carbon dioxide (scCO₂) as a green solvent was developed. The effects of cosolvent concentration (0–9% (w/w)), temperature (50–75 °C), and pressure (450–650 bar) were investigated with regard to the extraction yield. An optimized extraction was achieved with 9% (w/w) ethanol as a cosolvent, at 68 °C and 550 bar, allowing the extraction of 67.5 ± 3.7% of the cellular astaxanthin within 0.5 h. Prolonging the extraction time further increased the recovery to 93.3%, which is comparable to processes that have been established for the extraction of astaxanthin from microalgae and yeast. This approach provides a scalable and environmentally friendly solution for industrial astaxanthin recovery.

Keywords

Astaxanthin / Supercritical carbon dioxide / Extraction / Corynebacterium glutamicum / Engineering / Chemical Engineering

References

[1] AhmadkelayehS, CheemaSK, HawboldtK. Supercritical CO2 extraction of lipids and Astaxanthin from Atlantic shrimp by-products with static co-solvents: process optimization and mathematical modeling studies. J CO2 Utilization, 2022, 58: 101938.

[2] ÁlvarezCE, VardanegaR, Salinas-FuentesF, et al. . Effect of CO2 flow rate on the extraction of Astaxanthin and fatty acids from using supercritical fluid technology. Molecules, 2020, 25: 6044.

[3] AnY, KimT, ByeonH, et al. . Improved production of Astaxanthin from using a hybrid open–closed cultivation system. Appl Sci, 2024, 14: 1104.

[4] Astaxanthin Market (2024) Astaxanthin market size, share, growth & trends report 2030. In: Grand View Research. https://www.grandviewresearch.com/industry-analysis/global-astaxanthin-market. Accessed 28 Aug 2024

[5] BarreiroC, BarredoJL. Carotenoids production: A healthy and profitable industry. Methods in molecular biology, 2018New York, NYHumana45-55

[6] ChougleJA, BankarSB, ChavanPV, et al. . Supercritical carbon dioxide extraction of Astaxanthin from NBRC 101723: mathematical modelling study. Sep Sci Technol, 2016, 51: 2164-2173.

[7] de la FuenteJC, OyarzúnB, QuezadaN, del ValleJM. Solubility of carotenoid pigments (lycopene and astaxanthin) in supercritical carbon dioxide. Fluid Phase Equilib, 2006, 247: 90-95.

[8] Del ValleJM, UquicheEL. Particle size effects on supercritical CO2 extraction of oil-containing seeds. J Am Oil Chem Soc, 2002, 79: 1261-1266.

[9] DesaiRK, StreeflandM, WijffelsRH, EppinkMHM. Novel Astaxanthin extraction from using cell permeabilising ionic liquids. Green Chem, 2016, 18: 1261-1267.

[10] Di SanzoG, MehariyaS, MartinoM, et al. . Supercritical carbon dioxide extraction of Astaxanthin, Lutein, and fatty acids from microalgae. Mar Drugs, 2018, 16: 334.

[11] DiaoJ, SongX, ZhangL, et al. . Tailoring cyanobacteria as a new platform for highly efficient synthesis of Astaxanthin. Metab Eng, 2020, 61: 275-287.

[12] GalassoC, CorinaldesiC, SansoneC. Carotenoids from marine organisms: biological functions and industrial applications. Antioxidants, 2017, 6: 96.

[13] GöttlVL, MeyerF, SchmittI, et al. . Enhancing Astaxanthin biosynthesis and pathway expansion towards glycosylated C40 carotenoids by. Sci Rep, 2024, 14: 8081.

[14] HarithZT, LimaM, de CharalampopoulosA, ChatzifragkouD. Optimised production and extraction of Astaxanthin from the yeast. Microorganisms, 2020, 8: 430.

[15] HasunumaT, TakakiA, MatsudaM, et al. . Single-stage Astaxanthin production enhances the nonmevalonate pathway and photosynthetic central metabolism in Sp. PCC 7002. ACS Synth Biol, 2019, 8: 2701-2709.

[16] Hayashi M, Ishibashi T, Kuwahara D, Hirasawa K (2021) Commercial production of Astaxanthin with Paracoccus carotinifaciens. Advances in experimental medicine and biology. Springer, pp 11–20

[17] HenkeNA, WendischVF. Improved Astaxanthin production with by application of a membrane fusion protein. Mar Drugs, 2019, 17: 621.

[18] HenkeNA, HeiderSAE, Peters-WendischP, WendischVF. Production of the marine carotenoid Astaxanthin by metabolically engineered. Mar Drugs, 2016, 14: 1-21.

[19] HossainMS, Nik Ab RahmanNN, BalakrishnanV, et al. . Optimizing supercritical carbon dioxide in the inactivation of bacteria in clinical solid waste by using response surface methodology. Waste Manag, 2015, 38: 462-473.

[20] IrshadM, HongME, MyintAA, et al. . Safe and complete extraction of Astaxanthin from by efficient mechanical disruption of cyst cell wall. Int J Food Eng, 2019, 15: 20190128.

[21] KangCD, SimSJ. Direct extraction of Astaxanthin from culture using vegetable oils. Biotechnol Lett, 2008, 30: 441-444.

[22] KangX, MaoL, ShiJ, et al. . Supercritical carbon dioxide systems for sustainable and efficient dissolution of solutes: a review. Environ Chem Lett, 2024, 22: 815-839.

[23] KishimotoY, YoshidaH, KondoK. Potential anti-atherosclerotic properties of Astaxanthin. Mar Drugs, 2016, 14: 35.

[24] Knez Ž, Lütge C (2023) Industrial scale applications: Physical-based processes. Product, process and plant design using subcritical and supercritical fluids for industrial application 49–150. https://doi.org/10.1007/978-3-031-34636-1_3

[25] Knez Ž, Škerget M, KnezHrnčič M (2010) Principles of supercritical fluid extraction and applications in the food, beverage and nutraceutical industries. Separation, extraction and concentration processes in the food, beverage and nutraceutical industries. Woodhead Publishing, pp 3–38

[26] KnezMarkočičE, LeitgebM, et al. . Industrial applications of supercritical fluids: A review. Energy, 2014, 77: 235-243.

[27] KrichnavarukS, ShotiprukA, GotoM, PavasantP. Supercritical carbon dioxide extraction of Astaxanthin from with vegetable oils as co-solvent. Bioresour Technol, 2008, 99: 5556-5560.

[28] KumarS, KumarR, KumariA, PanwarA. Astaxanthin: A super antioxidant from microalgae and its therapeutic potential. J Basic Microbiol, 2022, 62: 1064-1082.

[29] LiJ, ZhuD, NiuJ, et al. . An economic assessment of Astaxanthin production by large scale cultivation of. Biotechnol Adv, 2011, 29: 568-574.

[30] Lim G, Bin, Lee SY, Lee EK et al (2002) Separation of Astaxanthin from red yeast Phaffia rhodozyma by supercritical carbon dioxide extraction. Biochemical engineering journal. Elsevier, pp 181–187

[31] LimKC, YusoffFM, ShariffM, KamarudinMS. Astaxanthin as feed supplement in aquatic animals. Rev Aquac, 2018, 10: 738-773.

[32] MachadoFRS, TrevisolTC, BoschettoDL, et al. . Technological process for cell disruption, extraction and encapsulation of Astaxanthin from. J Biotechnol, 2016, 218: 108-114.

[33] MachmudahS, ShotiprukA, GotoM, et al. . Extraction of Astaxanthin from using supercritical CO2 and ethanol as entrainer. Ind Eng Chem Res, 2006, 45: 3652-3657.

[34] MolinoA, MehariyaS, IovineA, et al. . Extraction of Astaxanthin and lutein from microalga in the red phase using CO2 supercritical fluid extraction technology with ethanol as co-solvent. Mar Drugs, 2018, 16: 432.

[35] MolinoA, RimauroJ, CasellaP, et al. . Extraction of Astaxanthin from microalga in red phase by using generally recognized as safe solvents and accelerated extraction. J Biotechnol, 2018, 283: 51-61.

[36] MonteroO, Macìas-SánchezMD, LamaCM, et al. . Supercritical CO2 extraction of β-carotene from a marine strain of the Cyanobacterium species. J Agric Food Chem, 2005, 53: 9701-9707.

[37] MooreWN, TaylorLT. Extraction and quantitation of Digoxin and acetyldigoxin from the leaf via near-supercritical methanol-modified carbon dioxide. J Nat Prod, 1996, 59: 690-693.

[38] NobreB, MarceloF, PassosR, et al. . Supercritical carbon dioxide extraction of Astaxanthin and other carotenoids from the microalga. Eur Food Res Technol, 2006, 223: 787-790.

[39] OtaS, MoritaA, OhnukiS, et al. . Carotenoid dynamics and lipid droplet containing Astaxanthin in response to light in the green Alga. Sci Rep, 2018, 8: 1-10.

[40] PanJL, WangHM, ChenCY, ChangJS. Extraction of Astaxanthin from by supercritical carbon dioxide fluid with ethanol modifier. Eng Life Sci, 2012, 12: 638-647.

[41] ParkSY, BinkleyRM, KimWJ, et al. . Metabolic engineering of for high-level Astaxanthin production with high productivity. Metab Eng, 2018, 49: 105-115.

[42] Picot-AllainC, MahomoodallyMF, AkG, ZenginG. Conventional versus green extraction techniques — a comparative perspective. Curr Opin Food Sci, 2021, 40: 144-156.

[43] PitaccoW, SamorìC, PezzolesiL, et al. . Extraction of Astaxanthin from with hydrophobic deep eutectic solvents based on oleic acid. Food Chem, 2022, 379: 132156.

[44] PraveenkumarR, LeeJ, VijayanD, et al. . Morphological change and cell disruption of cyst during high-pressure homogenization for Astaxanthin recovery. Appl Sci, 2020, 10: 513.

[45] PrimožičM, ČolnikM, KnezŽ, LeitgebM. Advantages and disadvantages of using scCO2 for enzyme release from halophilic fungi. J Supercrit Fluids, 2019, 143: 286-293.

[46] ReyesFA, MendiolaJA, IbañezE, Del ValleJM. Astaxanthin extraction from using CO2-expanded ethanol. J Supercrit Fluids, 2014, 92: 75-83.

[47] Rodríguez-SifuentesL, MarszalekJE, Hernández-CarbajalG, Chuck-HernándezC. Importance of downstream processing of natural Astaxanthin for pharmaceutical application. Front Chem Eng, 2020, 2: 601483.

[48] SaradaR, VidhyavathiR, UshaD, RavishankarGA. An efficient method for extraction of Astaxanthin from green Alga. J Agric Food Chem, 2006, 54: 7585-7588.

[49] SathasivamR, KiJS. A review of the biological activities of microalgal carotenoids and their potential use in healthcare and cosmetic industries. Mar Drugs, 2018, 16: 26.

[50] SeegerJ, WendischVF, HenkeNA. Extraction and purification of highly active Astaxanthin from fermentation broth. Mar Drugs, 2023, 21: 530.

[51] Stachowiak B, Szulc P (2021) Astaxanthin for the food industry. Molecules 26. https://doi.org/10.3390/molecules26092666

[52] ValderramaJO, PerrutM, MajewskiW. Extraction of Astaxantine and phycocyanine from microalgae with supercritical carbon dioxide. J Chem Eng Data, 2003, 48: 827-830.

[53] WangW, RaoL, WuX, et al. . Supercritical carbon dioxide applications in food processing. Food Eng Rev, 2021, 13: 570-591.

[54] WendischVF. Metabolic engineering advances and prospects for amino acid production. Metab Eng, 2020, 58: 17-34.

[55] WuT, HanB. Supercritical carbon dioxide (CO2) as green solvent. Green chemistry and chemical engineering, 2019New York, NYSpringer173-197.

[56] WuW, LuM, YuL. A new environmentally friendly method for Astaxanthin extraction from Xanthophyllomyces dendrorhous. Eur Food Res Technol, 2011, 232: 463-467.

[57] XieL, CahoonE, ZhangY, CiftciON. Extraction of Astaxanthin from engineered seed using ethanol-modified supercritical carbon dioxide. J Supercrit Fluids, 2019, 143: 171-178.

[58] YabuzakiJ. Carotenoids database: structures, chemical fingerprints and distribution among organisms. Database, 2017, 2017: 1-11.

[59] Youn HS, Roh MK, Weber A, Wilkinson GT, Chun BS (2007) Solubility of astaxanthin in supercritical carbon dioxide. Korean J Chem Eng 24:831–834. https://doi.org/10.1007/s11814-007-0049-z

[60] Zhuang Y, Zhu MJ (2021) Recent developments in Astaxanthin production from Phaffia rhodozyma and its applications. Global perspectives on Astaxanthin: from industrial production to food, health, and pharmaceutical applications. Academic, pp 225–251

Funding

Karlsruher Institut für Technologie (KIT) (4220)

RIGHTS & PERMISSIONS

The Author(s)