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Recovery of Astaxanthin from microalgae Using Simple and Energy-efficient Method

미세조류로부터의 에너지 효율적인 Astaxanthin 회수 기술 개발

  • Kim, Sun Young (Department of Advanced Materials & Chemical Engineering, Hannam University) ;
  • Oh, You-Kwan (Department of Chemical & Biomolecular Engineering, Pusan National University) ;
  • Ha, Sung Ho (Department of Advanced Materials & Chemical Engineering, Hannam University)
  • 김선영 (한남대학교 화공신소재공학과) ;
  • 오유관 (부산대학교 화공생명공학부) ;
  • 하성호 (한남대학교 화공신소재공학과)
  • Received : 2018.04.10
  • Accepted : 2018.05.08
  • Published : 2018.06.01

Abstract

The astaxanthin recovery efficiencies were compared in acetonitrile, acetone, methanol, dichloromethane : methanol (1:3, v/v) and ethylacetate : ethanol (1:1, v/v) as a extraction solvent after the grinding of the H. pluvialis cells. The astaxanthin extraction yield in acetone was 1.13~1.29 times higher than other extraction solvents. It was also found that 96.7% of astaxanthin accumulated in H. pluvialis could be recovered by a single extraction. Since astaxanthin exists mainly as astaxanthin esters in H. pluvialis, a gradient reversed-phase HPLC analysis was carried out for the separation of astaxanthin esters from the extracts of H. pluvialis. Among the astaxanthin inside the H. pluvialis cell, free astaxanthin was 45.9% and astaxanthin esters were the rest.

강력한 항산화물질인 astaxanthin의 함량이 다른 천연 공급원에 비해 높아 astaxanthin 생산균주로 주목받고 있는 Haematococcus pluvialis는 상당한 두께의 견고한 세포벽을 가지고 있어, 세포 파쇄를 위해 많은 에너지가 소모되고 비용이 비싼 방법들이 이용되고 있다. 이에 H. pluvialis로부터 막자와 막자사발을 이용하여 astaxanthin을 손쉽게 효율적으로 추출하는 방법을 제시하였다. 막자와 막자사발을 이용하여 분쇄한 후 추출용매로 acetonitrile, acetone, methanol, dichloromethane : methanol (1:3, v/v), ethylacetate : ethanol (1:1, v/v)로 사용하여 비교하였을 때, acetone을 이용하였을 때 astaxanthin을 1.13~1.29 배 더 높은 효율로 추출할 수 있었다. 또한 acetone으로 H. pluvialis로부터 추출할 경우, 1차 추출로 H. pluvialis에 축적되어 있는 전체 astaxanthin의 96.7%를 회수할 수 있을 정도로 acetone은 astaxanthin 추출효율이 높았다. H. pluvialis가 세포내에 축적하는 astaxanthin은 축적 특성상 ester-형태의 astaxanthin로 다량 축적하므로, 추출물 내의 다양한 형태의 astaxanthin을 분리하기 위하여 농도 구배 시스템을 적용한 HPLC 분석을 수행하였다. H. pluvialis에 축적되어 있는 전체 astaxanthin 중 free astaxanthin이 45.9%이고, 나머지 54.1%는 ester-형태의 astaxanthin이었다.

Keywords

References

  1. Hong, S. P., Kim, M. H. and Hwang, J. K., "Biological Functions and Production Technology of Cartenoids," J. Korean Soc. Food Sci. Nutr., 27, 1297-1306(1998).
  2. Guerin, M., Huntley, M. E. and Olaizola, M., "Haematococcus Astaxanthin; Applications for Human Health and Nutrition," Trends Biotechnol., 21(5), 210-216(2003). https://doi.org/10.1016/S0167-7799(03)00078-7
  3. Mann, V., Harker, M., Pecker, I. and Hirschberg, J., "Metabolic Engineering of Astaxanthin Production in Tobacco Flowers," Nature Biotechnol., 18, 888-892(2000). https://doi.org/10.1038/78515
  4. Han, D, Li, Y. and Hu, Q., "Astaxanthin in Microalgae: Pathways, Functions and Biotechnological Implications," Algae, 28, 131-147(2013). https://doi.org/10.4490/algae.2013.28.2.131
  5. Chumpolkulwong, N., Kakizono, T., Ishii, H., and Nishio, N., "Increased Astaxanthin Production by Phaffia rhodozyma Mutants Isolated as Resistant to Diphenylamine," J. Ferment. Bioeng., 83, 429-434(1997). https://doi.org/10.1016/S0922-338X(97)82996-0
  6. Chumpolkulwong, N., Kakizono, T., Ishii, H., and Nishio, N., "Enzymatic Conversion of Beta-carotene to Astaxanthin by Cellextracts of a Green Alga Haematococcus pluvialis," Biotechnol. Lett., 19, 443-446(1997). https://doi.org/10.1023/A:1018392010218
  7. Lim. G. B. Lee, S. Y., Lee, E. K., Haam, S. J. and Kim, W. S., "Separation of Astaxanthin from Red Yeast Phaffia rhodozyma by Supercritical Carbon Dioxide Extraction," J. Biochem. Eng., 11, 181-187(2002). https://doi.org/10.1016/S1369-703X(02)00023-2
  8. Lorenz, R. T. and Cysewski, G. R., "Commercial Potential for Haematococcus microalgae as a Natural Source of Astaxanthin," Trends Biotechnol., 18, 160-167(2000). https://doi.org/10.1016/S0167-7799(00)01433-5
  9. Schroeder, W. A. and Johnson, E. A., "Antioxidant Role of Carotenoids in Phaffia Rhodozyma," J. Gen. Microbiol., 39, 907-912 (1993).
  10. Kim, S., Cho, E., Yoo, J., In, M. -J. and Chae, H. J., "Extraction and Analysis of Astaxanthin from Haematococcus," J. Korean Soc. Food Sci. Nutr., 37, 1363-1368(2008). https://doi.org/10.3746/jkfn.2008.37.10.1363
  11. Hagen, C., Siegmund, S. and Braune, W., "Ultrastructural and Chemical Changes in the Cell Wall of Haematococcus pluvialis (Volvocales, Chlorophyta) during Aplanospore Formation," Eur. J. Phycol., 37, 217-226(2002). https://doi.org/10.1017/S0967026202003669
  12. Margalith, P. Z., "Production of Ketocarotenoids by Microalgae," Appl. Microbiol. Biotechnol., 51, 431-438(1999). https://doi.org/10.1007/s002530051413
  13. Boussiba, S., "Carotenogenesis in the Green Alga Haematococcus pluvialis: Cellular Physiology and Stress Response," Physiol. Plant, 108, 111-117(2000). https://doi.org/10.1034/j.1399-3054.2000.108002111.x
  14. Fabregas, J., Dominguez, A., Maseda, A. and Otero, A., "Interactions Between Irradiance and Nutrient Availability During Astaxanthin Accumulation and Degradation in Haematococcus pluvialis," Appl. Microbiol. Biotechnol., 61, 545-551(2003). https://doi.org/10.1007/s00253-002-1204-4
  15. Sarada, R., Tripathi, U. and Ravishankar, G. A., "Influence of Stress on Astaxanthin Production in Haematococcus pluvialis Grown under Different Culture Conditions," Process Biochem., 37, 623-627(2002). https://doi.org/10.1016/S0032-9592(01)00246-1
  16. Kim, D. Y., Vijayan, D., Praveenkumar, R., Han, J. -I., Lee, K., Park, J. Y., Chang, W.S., Lee, J. -S. and Oh, Y. -K., "Cell-wall Disruption and Lipid/astaxanthin Extraction from Microalgae: Chlorella and Haematococcus," Bioresour. Technol., 199, 300-310(2016). https://doi.org/10.1016/j.biortech.2015.08.107
  17. Wellburn, A. R., "The Spectral Determination of Chlorophylls a and b, as well as Total Carotenoids, Using Various Solvents with Spectrophotometers of Different Resolution," J. Plant Physiol., 144, 307-313(1994). https://doi.org/10.1016/S0176-1617(11)81192-2
  18. Yuan, J. P. and Chen, F., "Chromatographic Separation and Purification of trans-Astaxanthin from the Extracts of Haematococcus pluvialis," J. Agric. Food Chem., 46, 3371-3375(1998). https://doi.org/10.1021/jf980039b
  19. Jo, J., Shin, S., Jung, H., Min, B., Kim, S. and Kim, J., "Process Development for Production of Antioxidants from Lipid Extracted Microalgae Using Ultrasonic-assisted Extraction," Korean Chem. Eng. Res., 55, 542-547(2017).
  20. Kim, J. and Ha, S. H., "Hydrothermal Pretreatment of Ulva pertusa Kjellman Using Microwave Irradiation for Enhanced Enzymatic Hydrolysis," Korean Chem. Eng. Res., 53, 570-575(2015). https://doi.org/10.9713/kcer.2015.53.5.570
  21. Johnson, E. A. and An, G. H., "Astaxanthin from Microbial Sources," Crit. Rev. Biotechnol., 11, 297-326(1991). https://doi.org/10.3109/07388559109040622
  22. Yuan, J. P. and Chen, F., "Purification of trans-Astaxanthin from a High-yielding Astaxanthin Ester-producing Strain of the Microalga Haematococcus pluvialis," Food Chem., 68, 443-448(2000). https://doi.org/10.1016/S0308-8146(99)00219-8
  23. Orosa, M., Franqueira, D., Cid, A. and Abalde, J., "Analysis and Enhancement of Astaxanthin Accumulation in Haematococcus pluvialis," Bioresour. Technol., 96, 373-378(2005). https://doi.org/10.1016/j.biortech.2004.04.006
  24. Kim, Z. H., Kim, S. H., Lee, H. S. and Lee, C. G., "Enhanced Production of Astaxanthin by Flashing Light Using Haematococcus pluvialis," Enzyme Microb. Technol., 39, 414-419(2006). https://doi.org/10.1016/j.enzmictec.2005.11.041