GOVPH
MENU

Preliminary Investigation of the Carotenoid Composition
of Erythrobacter sp. Strain KJ5 by High-Performance
Liquid Chromatography and Mass Spectrometry

Ayu Dita Juliadiningtyas1, Delianis Pringgenies1, Heriyanto2,3,
Katarina Purnomo Salim2, Ocky Karna Radjasa1, Yuzo Shioi2,
Leenawaty Limantara2,4, and Tatas Hardo Panintingjati Brotosudarmo2*

1Faculty of Fisheries and Marine Science, Diponegoro University,
 Jl. Prof. Soedarto Tembalang, Semarang-50275, Indonesia
2Ma Chung Research Center for Photosynthetic Pigments, Ma Chung University,
Villa Puncak Tidar N-1, Malang-65151, Indonesia
3Deparment of Plant Physiology and Biochemistry, Jagiellonian University,
ul. Gronostajowa 7, 30-387 Krakow, Poland
4Center for Urban Studies, Universitas Pembangunan Jaya,
Jl. Cendrawasih Raya B7/P, South Tangerang-15413, Banten, Indonesia

*Corresponding author: This email address is being protected from spambots. You need JavaScript enabled to view it.

 

 

ABSTRACT

Separation and identification of carotenoids (Cars) from aerobic marine bacterium Erythrobacter sp. strain KJ5 are reported. The cells of Erythrobacter sp. were grown in a Shioi medium at 28.5° C for three days. Among the four solvents tested, the mixture of methanol and acetone (3:7, v/v) was determined as the optimum solvent for Car extraction from the cells by measuring its absorption spectrum. The Cars were separated via reversed-phase high-performance liquid chromatography using a C8 column and identified by a UV-Vis photodiode array detector and an electrospray ionization mass spectrometry. Bacteriochlorophyll a was not detected from the extracts of cells grown under both light and dark conditions. At least 16 peaks of Cars were separated, wherein eleven peaks showed the same absorption spectrum with λmax at 452-453 nm and at 478-480 nm. The other five peaks had an additional absorption peak at 340 nm, which belongs to cis-isomeric form. Two peaks of Cars were identified to be zeaxanthin and β-carotene.

Key words: β-carotene, carotenoids, Erythrobacter sp., high-performance liquid chromatography, mass spectrometry, zeaxanthin

 

INTRODUCTION

Carotenoids (Cars) are isoprenoid pigments in the form of yellow, orange, and red color found in both photosynthetic and non-photosynthetic organisms. Cars play a critical role in the photosynthetic process for higher plants, algae, and anoxygenic photosynthetic bacteria. Moreover, Cars act as photoprotector by quenching the chlorophyll triplet and by scavenging singlet oxygen to protect cellular constituents i.e., DNA and membranes (Frank & Cogdell 1996). In addition, many evidences reveal the beneficial health effects of Cars as antiobesity (Miyashita 2009), antioxidant (Yeum et al. 2009), and anticancer (Kotake-Nara et al. 2001) agents. . . . . read more

 

REFERENCES

AMANN R, LUDWIG W, SCHLEIFER K. 1995. Phylogenetic identification and in situ detection of individual microbial cells without cultivation. Microbiological Reviews 59:143-169.
BRITTON G. 1995. UV/Visible spectroscopy. In: Carotenoids. Vol. 1B: Spectroscopy. Britton G, Liaaen-Jensen S, Pfander H eds. Basel: Birkhäuser Verlag. p. 13-62.
BROTOSUDARMO THP, HERIYANTO, SHIOI Y, INDRIATMOKO, ADHIWIBAWA MAS, INDRAWATI R, LIMANTARA L. 2017. Composition of main dominant pigments from potentially commercial two edible seaweeds. Philippine Journal of Science (In Press).
ENZELL RC, BACK S. 1995.  Mass spectrometry. In: Carotenoids Vol. 1B: Spectroscopy. Britton G, LIAAEN-JENSEN S & Pfander H eds. Basel: Birkhäuser Verlag. p. 261-320.
FIEDOR L, HERIYANTO, FIEDOR J, PILCH M. 2016. Effects of molecular symmetry on the electronic transitions in carotenoids. The Journal of Physical Chemistry Letters 7:1821-1829.
FRANK HA, COGDELL RJ. 1996. Carotenoids in photosynthesis. Photochemistry and Photobiology 63:257-64.
GROSS J. 1991. Pigments in vegetables: chlorophylls and carotenoids. New York: Van Nostrand Reinhold. 351p.  
HERIYANTO, JULIADININGTYAS AD, SHIOI Y, LIMANTARA L, BROTOSUDARMO THP. 2017. Analysis of pigment composition of brown seaweeds collected from Panjang Island, Central Java, Indonesia. Philippine Journal of Science 146(3):323-330.
KOTAKE-NARA E, KUSHIRO M, ZHANG H, SUGAWARA T, MIYASHITA K, NAGAO A. 2001. Carotenoids affect proliferation of human prostate cancer cells. Journal of Nutrition 131:3303-3306.
KOYAMA Y, FUJII R. 1999. Cis-trans carotenoids in photosynthesis: Configurations, excited-state properties and physiological functions. In:  The photochemistry of carotenoids. Frank HA, Young AJ, Britton G, & Cogdell RJ eds. The Netherlands: Kluwer Academic Publishers. p. 161-188.
LIMANTARA L, HERIYANTO. 2011. Photostability of bacteriochlorophyll a and its derivatives as potential sensitizers for photodynamic cancer therapy: The study on acetone-water and methanol-water solvents. Indonesian Journal of Chemistry 11:154-162.
MARASCO EK. 2008. Identification and characterization of bacterial carotenoids cleavage oxygenases. [MS dissertation]. Minnesota: University of Minnesota. 222p.  
MISAWA N. 2011. Carotenoid β-ring hydroxylase and ketolase from marine bacteria-promiscuous enzymes for synthesizing functional xanthophylls. Marine Drugs 9:757-771.
MIYASHITA K. 2009. The carotenoid–fucoxanthin from brown seaweed affects obesity. Lipid Technology 21:186-190.
MORTENSEN A. 2006. Carotenoids and other pigments as natural colorants. Pure Applied Chemistry 78:1477-1491.
RIVERA SM, VILARó F, CANELA R. 2011. Determination of carotenoids by liquid chromatography/mass spectrometry: Effect of several dopants. Analytical and Bioanalytical Chemistry 400:1339-1346.
RIVERA SM, CHRISTOU P, CANELA-GARAYOA R. 2014. Identification of carotenoids using mass spectrometry. Mass Spectrometry Reviews 33:353-372.
RODRIGUEZ-AMAYA DB. 2001. A guide to carotenoid analysis in foods. Washington, DC: ILSI Press.
ROUSEFF R, RALEY L, HOFSOMMER H. 1996. Application of diode array detector with a C-30 reversed phase column for the separation and identification of saponified orange juice carotenoids. Journal of Agricultural and Food Chemistry 44:2176-2181.
SCHIEDT K & LIAAEN-JENSEN S. 1995. Isolation and analysis. In: Carotenoids. Volume 1A: Isolation and analysis. Britton G, Liaaen-Jensen S, Pfander H eds.  Basel: Birkhäuser Verlag. p. 81-108.  
SCOTTER M. 2009. The chemistry and analysis of annatto food colouring: a review. Food Additives and Contaminants 26:1123-1145.
SHIBA T, SIMIDU U. 1982. Erythrobacter longus gen. nov., sp. nov., an aerobic bacterium which contains bacteriochlorophyll a. International Journal of Systematic Bacteriology 32:211-217.
SHINDO K, MISAWA N. 2014. New and rare carotenoids isolated from marine bacteria and their antioxidant activities. Marine Drugs 12:1690-98.
SHIOI Y. 1986. Growth characteristics and substrate specificity of aerobic photosynthetic bacterium, Erythrobacter sp. (OCh 114). Plant & Cell Physiology 27:567-572.
SHIOI Y, DOI M. 1988. Control of bacteriochlorophyll accumulation by light in an aerobic photosynthetic bacterium, Erythrobacter sp. OCh114. Archives of Biochemistry and Biophysics 266:470-477.
SOWMYA R, SACHINDRA NM. 2015. Carotenoid production by Formosa sp. KMW, a marine bacteria of Flavobacteriaceae family: Influence of culture conditions and nutrient composition. Biocatalysis and Agricultural Biotechnology 4:559-567.
STAFSNES MH, JOSEFSEN KD, KILDAHL-ANDERSEN G, VALLA S, ELLINGSEN TE, BRUHEIM P. 2010. Isolation and characterization of marine pigmented bacteria from Norwegian coastal waters and screening for carotenoids with UVA-blue light absorbing properties. Journal of Microbiology 48:16-23.
TANG G, RUSSELL RM. 2009. Carotenoids as provitamin A. In: Carotenoids, Vol. 5: Nutrition and Health. Britton G, Liaaen-Jensen S, & Pfander H eds. Basel: Birkhäuser Verlag. p. 149-172.
VAN BREEMEN RB. 1995. Electrospray liquid chromatography-mass spectrometry of carotenoids. Analytical Chemistry 67:2004-2009.
WUSQY NK, LIMANTARA L, KARWUR FF. 2014. Exploration, isolation and quantification of β-carotene from bacterial symbiont of Acropora sp. Microbiology Indonesia 8:58-64.
YEUM KJ, ALDINI G, RUSSELL RM, KRINSKY NI. 2009. Antioxidant/pro-oxidant action of carotenoids. In: Carotenoids, Vol. 5: Nutrition and Health. Britton G, Liaaen-Jensen S, & Pfander H eds. Basel: Birkhäuser Verlag. p. 235-268.
YOON JH, KIM H, KIM IG, KANG KH, PARK YH. 2003. Erythrobacter flavus sp. nov., a slight halophile from the East Sea in Korea. International Journal of Systematic and Evolutionary Microbiology 53:1169-74.
ZAPATA M, RODRIGUEZ F, GARRIDO JL. 2000. Separation of chlorophylls and carotenoids from marine phytoplankton: a new HPLC method using a reversed phase C8 column and pyridine-containing mobile phases. Marine Ecology Progress Series 195:29-45.
ZHENG Q, KOBLIZEK M, BEATTY JT, JIAO N. 2013. Evolutionary divergence of marine aerobic anoxygenic phototrophic bacteria as seen from diverse organisations of their photosynthesis gene clusters. Advances in Botanical Research 66:359-383.
ZOBELL CE, FELTHAM CB. 1934. Preliminary studies on the distribution and characteristics of marine bacteria. Bulletin of the Scripps Institution of Oceanography of the University of California, Technical series 3:279-296.