Morphological, Molecular, and Biochemical Characterization of Astaxanthin-Producing Green Microalga Haematococcus sp. KORDI03 (Haematococcaceae, Chlorophyta) Isolated from Korea
0
조회
0
다운로드
3
인용
0
참고문헌
키워드
초록
A unicellular red microalga was isolated from environmental freshwater in Korea, and its morphological, molecular, and biochemical properties were characterized. Morphological analysis revealed that the isolate was a unicellular biflagellated green microalga that formed a non-motile, thick-walled palmelloid or red aplanospore. To determine the taxonomical position of the isolate, its 18S rRNA and rbcL genes were sequenced and phylogenetic analysis was performed. We found that the isolate was clustered together with other related Haematococcus strains showing differences in the rbcL gene. Therefore, the isolated microalga was classified into the genus Haematococcus, and finally designated Haematococcus sp. KORDI03. The microalga could be cultivated in various culture media under a broad range of pH and temperature conditions. Compositions of the microalgal cellular components were analyzed, and its protein, carbohydrate, and lipid compositions were estimated to be 21.1 ± 0.2%, 48.8 ± 1.8%, and 22.2 ± 0.9%, respectively. In addition, D-glucose and D-mannose were the dominant monosaccharides in the isolate, and its amino acids were composed mainly of aspartic acid, glutamic acid, alanine, and leucine. Moreover, several polyunsaturated fatty acids accounted for about 80% of the total fatty acids in Haematococcus sp. KORDI03, and the astaxanthin content in the red aplanospores was estimated to be 1.8% of the dry cell weight. To the best of our knowledge, this is the first report of an Haematococcus sp. isolated from Korea, which may be used for bioresource production in the microalgal industry.
Abstract
A unicellular red microalga was isolated from environmental freshwater in Korea, and its morphological, molecular, and biochemical properties were characterized. Morphological analysis revealed that the isolate was a unicellular biflagellated green microalga that formed a non-motile, thick-walled palmelloid or red aplanospore. To determine the taxonomical position of the isolate, its 18S rRNA and rbcL genes were sequenced and phylogenetic analysis was performed. We found that the isolate was clustered together with other related Haematococcus strains showing differences in the rbcL gene. Therefore, the isolated microalga was classified into the genus Haematococcus, and finally designated Haematococcus sp. KORDI03. The microalga could be cultivated in various culture media under a broad range of pH and temperature conditions. Compositions of the microalgal cellular components were analyzed, and its protein, carbohydrate, and lipid compositions were estimated to be 21.1 ± 0.2%, 48.8 ± 1.8%, and 22.2 ± 0.9%, respectively. In addition, D-glucose and D-mannose were the dominant monosaccharides in the isolate, and its amino acids were composed mainly of aspartic acid, glutamic acid, alanine, and leucine. Moreover, several polyunsaturated fatty acids accounted for about 80% of the total fatty acids in Haematococcus sp. KORDI03, and the astaxanthin content in the red aplanospores was estimated to be 1.8% of the dry cell weight. To the best of our knowledge, this is the first report of an Haematococcus sp. isolated from Korea, which may be used for bioresource production in the microalgal industry.
참고문헌 (38)
- Allen MM, "Simple conditions for growth of unicellular blue-green algae on plates", J. Phycol, 4, pp.1~4 (1968)
- AOAC, "Official Methods of Analysis of the Association of Official Analytical Chemists" (2006), AOAC International
- Bakker FT, "The Cladophora complex(Chlorophyta) : new views based on 18S rRNA gene sequences. Mol. Phylogenet. Evol. 3 : 365-382", Mol. Phylogenet. Evol, 3, pp.365~382 (1994)
- Bar E, "Pigment and structural changes in Chlorella zofingiensis upon light and nitrogen stress", J. Plant Physiol, 146, pp.527~534 (1995)
- Bligh EG, "A rapid method of total lipid extraction and purification", Can. J. Biochem. Physiol, 37, pp.911~917 (1959)
- Bon J, "Isolation of astaxanthinoverproducing mutants of Phaffia rhodozyma", Biotechnol. Lett, 19, pp.109~112 (1997)
- Boussiba S, "Changes in pigments profile in the green alga Haematococcus pluvialis exposed to environmental stresses", Biotechnol. Lett, 21, pp.601~604 (1999)
- Brown MR, "Biochemical composition of microalgae from the green algal classes Chlorophyceae and Prasinophyceae. 1. Amino acids, sugars and pigments", J. Exp. Mar. Biol. Ecol, 161, pp.91~113 (1992)
- Buchheim MA, "Phylogeny of the colonial green flagellates: a study of 18S and 2 6S rRNA sequence data", Biosystems, 25, pp.85~100 (1991)
- Buchheim MA, "The blood alga: phylogeny of Haematococcus (Chlorophyceae)inferred from ribosomal RNA gene sequence data", Eur. J. Phycol, 48, pp.318~329 (2013)
- Choubert G, "Carotenoid pigments of the green alga Haematococcus pluvialis: assay on rainbow trout, Oncorhynchus mykiss, pigmentation in comparison with synthetic astaxanthin and canthaxanthin", Aquaculture, 112, pp.217~226 (1993)
- Damiani MC, "Lipid analysis in Haematococcus pluvialis to assess its potential use as a biodiesel feedstock", Bioresour. Technol, 101, pp.3801~3807 (2010)
- Demchenko E, "Generic and species concepts in Microglena (previously the Chlamydomonas monadina group) revised using an integrative approach", Eur. J. Phycol, 47, pp.264~290 (2012)
- Denery JR, "Pressurized fluid extraction of carotenoids from Haematococcus pluvialis and Dunaliella salina and kavalactones from Piper methysticum", Anal. Chim. Acta, 501, pp.175~181 (2004)
- Fraser PD, "In vitro characterization of astaxanthin biosynthetic enzymes", J. Biol. Chem, 272, pp.6128~6135 (1997)
- Gobantes I, "Serum carotenoid concentration changes during sexual maturation in farmed rainbow trout (Oncorhynchus mykiss)", J. Agric. Food Chem, 46, pp.383~387 (1998)
- Grung M, "Algal carotenoids 51. Secondary carotenoids 2. Haematococcus pluvialis aplanospores as a source of (3S, 3’S)-astaxanthin esters", J. Appl. Phycol, 4, pp.165~171 (1992)
- Guindon S, "PHYML Online — a web server for fast maximum likelihood-based phylogenetic inference", Nucleic Acids Res, 33, pp.557~559 (2005)
- Hagen C, "Ultrastructural and chemical changes in the cell wall of Haematococcus pluvialis (Volvocales, Chlorophyta) during aplanospore formation", Eur. J. Phycol, 37, pp.217~226 (2002)
- Hanyuda T, "Phylogenetic relationships within Cladophorales (Ulvophyceae, Chlorophyta) inferred from 18S rRNA gene sequences, with special reference to Aegagropila linnaei", J. Phycol, 38, pp.564~571 (2002)
- Harker M, "Autotrophic growth and carotenoid production of Haematococcus pluvialis in a 30liter air-lift photobioreactor", J. Ferment. Bioeng, 82, pp.113~118 (1996)
- Harris EH, "The Chlamydomonas Sourcebook: A Comprehensive Guide to Biology and Laboratory Use" (1989), Academic Press
- Huelsenbeck JP, "MRBAYES: Bayesian inference of phylogenetic trees", Bioinformatics, 17, pp.754~755 (2001)
- Johnson EA, "Astaxanthin from microbial sources", Crit. Rev. Biotechnol, 11, pp.297~326 (1991)
- Tatyana A. Klochkova, "Cold-tolerant strain of Haematococcus pluvialis (Haematococcaceae, Chlorophyta) from Blomstrandhalvøya (Svalbard)", ALGAE, 28(2), pp.185~192 (2013), 한국조류학회I[DOI]
- Knothe G, "“Designer” biodiesel: optimizing fatty ester composition to improve fuel properties", Energy Fuels, 22, pp.1358~1364 (2008)
- A technical review of Haematococcus algae, "Lorenz RT", Cyanotech Corporation, Kailua-Kona, 1~12, pp.NatuRoseTM Technical Bulletin #060 (1999)
- Lorenz RT, "Commercial potential for Haematococcus microalgae as a natural source of astaxanthin", Trends Biotechnol, 18, pp.160~167 (2000)
- Nakada T, "Molecular systematics of Volvocales (Chlorophyceae, Chlorophyta) based on exhaustive 18S rRNA phylogenetic analyses", Mol. Phylogenet. Evol, 48, pp.281~291 (2008)
- Nichols HW, "Handbook of Phycological Methods", pp.7~24 (1973), Cambridge University Press
- Nichols PD, "Determination of monosaturated fatty acid double-bond position and geometry for microbial monocultures and complex consortia by capillary GC-MS of their dimethyl disulphide adducts", J. Microbiol. Methods, 5, pp.49~55 (1986)
- Parajó JC, "Production of carotenoids by Phaffia rhodozyma growing on media made from hemicellulosic hydrolysates of Eucalyptus globulus wood", Biotechnol. Bioeng, 59, pp.501~506 (1998)
- Pröschold T, "Molecular phylogeny and taxonomic revision of Chlamydomonas (Chlorophyta). I. Emendation of Chlamydomonas Ehrenberg and Chloromonas Gobi, and description of Oogamochlamys gen", nov. and Lobochlamys gen. nov. Protist, 152, pp.265~300 (2001)
- Spurr AR, "A low-viscosity epoxy resin embedding medium for electron microscopy", J. Ultrastruct. Res, 26, pp.31~43 (1969)
- Stansell G, "Microalgal fatty acid composition: implications for biodiesel quality", J. Appl. Phycol, 24, pp.791~801 (2012)
- Wayama M, "Three-dimensional ultrastructural study of oil and astaxanthin accumulation during encystment in the green alga Haematococcus pluvialis", PLoS ONE, 8, pp.e53618~ (2013)
- Yokoyama A, "Composition and presumed biosynthetic pathway of carotenoids in the astaxanthinproducing bacterium Agrobacterium aurantiacum", FEMS Microbiol. Lett, 128, pp.139~144 (1995)
- Yuan JP, "Purification of trans-astaxanthin from a high-yielding astaxanthin ester-producing strain of the microalga Haematococcus pluvialis", Food Chem, 68, pp.443~448 (2000)