Optimization of Complex Media for the Production of Lutein from Chlorella Pyrenoidosa

Transcription

1 International Journal of Biotechnology and Bioengineering Research. ISSN -8, Volume, Number (), pp. 8-9 Research India Publications ijbbr.htm Optimization of Complex Media for the Production of Lutein from Chlorella Pyrenoidosa B. Shashee and U.S. Annapure Food and Engineering Department, Institute of Chemical Technology, Matunga, Mumbai-9. Abstract Lutein is a xanthophyll which is known to contain oxygen. It is the major pigment present in the macula of the eyes. It is also known to have certain food applications like pigmentation agent in animal tissue and products and also for the coloration of the food, drugs and cosmetics. Microalgae strain Chlorella pyrenoidosa is used for the production of lutein. Different media (Basal, Knop s, BG and Allen and Arnone) are screened for the heterotrophic production of lutein from the C.pyrenoidosa. The strain is chosen for further investigation as it has reported higher biomass as well as production of lutein. All the media supported the growth of microalgae and lutein formation but higher biomass and lutein concentration was achieved in the Allen and Arnone medium containing glucose. After which A and A media was optimized for enhancing the growth and lutein production. Keywords: Lutein, media optimization, microencapsulation, Chlorella.. Introduction Carotenoids are compounds that can be found in higher plants and algae, as well as in non-photosynthetic organisms like animals (although they are not able to synthesize carotenoids), fungi, and bacteria, which are responsible for the red, orange and, yellow colors of plant leaves, fruits, and flowers, as well as for the color of feathers, crustacean shells, fish flesh and skin, etc. Lutein is one such carotenoid which functions as an antioxidant and has gathered increasing attention due to its potential role in preventing age related macular degeneration. It is also being widely used for the pigmentation of animal tissues and products, as well as for the coloration of foods,

2 8 B. Shashee & U.S. Annapure drugs and cosmetics. Lutein is recently also gaining importance as nutraceutical compound against macular degeneration and also play an important role in maintaining a normal visual function. It is also been reported to prevent certain type of cancers and protect skin against UV-induced damage. Marigold flower petals are known to be excellent source of lutein as they contain high level of lutein and no significant level of other carotenoids. But continuous reports of lutein producing microalgae pose question is these microorganism can become an alternative source. Microalgae thus have been recognized as promising source for lutein and increasing the interest of companies for microalgae as a natural source of products. More recently, significant attention has been drawn to the use of microalgae as a source of feedstuff for animals due to the accumulation of high value nutrients in the cells, such as astaxanthin and xanthophylls. Lutein is not only one of the most prominent carotenoids in human serum and foods, but also the representative of, carotenoids. Lutein has been used for the pigmentation of animal tissues and products, as well as for the coloration of foods, drugs and cosmetics. Lutein is largely consumed as food colorant and its sales amount to $,, in the US only. Micro algal production of high value added products particularly carotenoids for human health has gained importance in recent years. The food, feed, pharamaceuticals, cosmetic and chemical industries benefit from microalgal products. several microalgae have been considered as potential sources of lutein/astaxanthin for several reasons: (a) its high lutein content (..% dry weight) compared to marigold petals; (b) petals do not have to be separated and the whole microalgal biomass is processed; (c) a homogeneous biomass is produced at a constant rate regardless of time and weather, so it lends itself better to a precisely designed extraction process; (d) valuable by-products that can be used to produce protein hydrolysates, other pigments, and even valuable lipids depending on the strain. Chlorella pyrenoidosa was chosen as microalgae as it is known to grow under autotrophic, heterotrophic and mixotrophic condition, and thus aim of the study was to investigate the media which showed highest biomass and lutein production and in future scope is to investigate the effect of complex media and optimization for enhancing the lutein production.. Materials and Methods. Microorganism Chlorella pyrenoidosa NCIM 8 from NCL Pune.. Media Basal media, Kuhl s media, Allen and Arnone media and BG media.

3 Optimization of Complex Media for the Production of Lutein from Chlorella 8. Culture condition The strain of Chlorella pyrenoidosa was grown in ml of different liquid media having ph of. each for hrs at C, rpm under /8 hrs light( watts tubelight) and dark period.. Lutein was extracted using standard method and analysed using HPLC. Media optimization by one factor method Table : Optimization conditions for one factor method. Incubation time to hrs Seed age to hrs Inoculums size -% v/v of 9 hrs old culture Effect of temperature,, and C Effect of ph ph from -9 Effect of different carbon source Glucose, fructose, sucrose, glycerol, galactose Effect of nitrogen source Urea, ammonium carbonate, KNO, NaNO, Yeast extract. Results and Discussions a) The media used for screening for production of lutein were Basal, Kuhl, A and A and BG. The strain Chlorella pyrenoidosa was grown autotrophically under defined conditions. The Arnone and BG media showed highest production of lutein i.e 8+ µg/l. The results for screening of media are shown in figure.. 8 Basal Kuhl Allen and Arnone BG.... Figure.: Media screening for lutein production.

4 8 B. Shashee & U.S. Annapure b) Optimization of medium by one factor method The production of lutein was optimum at 8 th h (+.µg/l) while DCW was around.98+.gm/l as shown in figure.. In case of effect of seed age maximum lutein production was observed in 9h old culture which was µg/L with DCW of.8+.gm/l (figure.). When inoculums size was around %v/v of 9 h old culture maximum lutein production was observed about +.8µg/L (figure.). Optimization of temperature(figure.) and ph (figure.) was also done where, in case of temperature maximum lutein production was observed at C with 9+.8µg/L of lutein and DCW of.98+.gm/l, while in ph maximum production was obtained at ph of with 8µg/L of lutein. When various carbon and nitrogen sources were used highest lutein production was found to be in glucose and urea, with 89+ µg/l lutein in glucose (figure.) and 9+ µg/l lutein in urea (figure.8) Figure.: Effect of incubation time (maximum at th day). 8 Figure.: Effect of seed age (maximum with days seed age).

5 Optimization of Complex Media for the Production of Lutein from Chlorella 8 Figure.: Effect of inoculum size(maximum % dilution). 8 Figure.: Effect of temperature(maximum at C) Figure.: Effect of ph (maximum at ph)

6 88 B. Shashee & U.S. Annapure 8 Figure.: Effect of carbon sources(maximum in glucose) 8 Figure.8: Effect of nitrogen sources(maximum in urea). Note:- Each graph denotess Y and Y as lutein in µg/l and DCW in gm/l, while X axis is the factor which is being optimized like in fig. media, fig. incubation time, fig. seed age, fig. inoculum size, fig. temperture, figure. ph, fig. carbon source and fig.8 nitrogen source.. Conclusion Various medias like Kuhl, Basal, BG and A and A medium were used for screening for lutein production and amongst these Arnone medium was found to be best for lutein production, followed by BG. Media was optimized for the maxumum production of lutein.

7 Optimization of Complex Media for the Production of Lutein from Chlorella 89. Future Scope Synthetic media is expensive thus fruit waste and other waste like molasses can be used for the production of lutein which would eliminate the use of glucose as energy and carbon source and thus utilize the waste as the carbon as well as nitrogen source. References [] Bial, A. M. & Physiology, C. Accumulation of astaxanthin and lutein in Chlorella zofingiensis ( Chlorophyta ) (). [] Casal, C., Cuaresma, M., Vega, J. M. & Vilchez, C. Enhanced productivity of a lutein-enriched novel acidophile microalga grown on urea. Marine drugs 9, 9 (). [] Cordero, B. F. et al. Enhancement of lutein production in Chlorella sorokiniana (Chorophyta) by improvement of culture conditions and random mutagenesis. Marine drugs 9, (). [] Fernández-Sevilla, J. M., Acién Fernández, F. G. & Molina Grima, E. Biotechnological production of lutein and its applications. Applied Microbiology and Biotechnology 8, (). [] Kim, M. K. et al. Enhanced production of Scenedesmus spp. (green microalgae) using a new medium containing fermented swine wastewater. Bioresource technology 98, 8 (). [] Rao, R., Sarada, A., Ravishankar, R. & Aswathanarayana, G. Enhancement of Carotenoids in Green Alga-Botryococcus braunii in Various Autotrophic Media under Stress Conditions. () [] Sánchez, J. F. et al. Biomass and lutein productivity of Scenedesmus almeriensis: influence of irradiance, dilution rate and temperature. Applied microbiology and biotechnology 9, 9 9 (8). [8] Sánchez, J. F. et al. Influence of culture conditions on the productivity and lutein content of the new strain Scenedesmus almeriensis. Process Biochemistry, 98 (8). [9] Shen, Y., Pei, Z., Yuan, W. & Mao, E. Effect of nitrogen and extraction method on algae lipid yield., (9). [] Shi, X., Chen, F., Yuan, J. & Chen, H. Heterotrophic production of lutein by selected Chlorella strains. (99). [] Shi, X., Zhang, X. & Chen, F. Heterotrophic production of biomass and lutein by Chlorella protothecoides on various nitrogen sources. Enzyme and microbial technology, 8 (). [] Sujith, A. P. A., Hymavathi, T. V & Devi, P. Y. Supercritical Fluid Extraction of Lutein Esters from Marigold Flowers and their Hydrolysis by Improved Saponification and Enzyme Biocatalysis. ().

8 9 B. Shashee & U.S. Annapure [] Wu, Z., Qu, C. & Shi, X. Biochemical System Analysis of Lutein Production by Heterotrophic Chlorella pyrenoidosa in a Fermentor. 98, (9).