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Ultraviolet and 5′Fluorodeoxyuridine Induced Random Mutagenesis in Chlorella vulgaris and Its Impact on Fatty Acid Profile: A New Insight on Lipid-Metabolizing Genes and Structural Characterization of Related Proteins

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Abstract

The present study was aimed at randomly mutating the microalga, Chlorella vulgaris, in order to alter its cellular behaviour towards increased lipid production for efficient biodiesel production from algal biomass. Individual mutants from ultraviolet light (UV-1 (30 s exposure), UV-2 (60 s exposure) and UV-3 (90 s exposure)) and 5′fluorodeoxyuridine (5′FDU-1 (0.25 mM) and 5′FDU-2 (0.50 mM)) exposed cells were identified to explore an alternative method for lipid enhancement. A marginally significant decrease in biomass in the UV mutants; marked increase in the lipid content in UV-2 and 5′FDU-1 mutants; significant increase in saturated fatty acids level, especially in UV-2 mutant; insignificant increase in lipid production when these mutants were subjected to an additional stress of nitrogen starvation and predominantly enhanced level of unsaturated fatty acids in all the strains except UV-2 were noted. Chloroplast ultrastructural alterations and defective biosynthesis of chloroplast specific lipid constituents were observed in the mutants. Modelling of three-dimensional structures of acetyl coA carboxylase (ACCase), omega-6, plastid delta-12 and microsomal delta-12 fatty acid desaturases for the first time and ligand-interaction studies greatly substantiated our findings. A replacement of leucine by a serine residue in the acetyl coA carboxylase gene of UV-2 mutant suggests the reason behind lipid enhancement in UV-2 mutant. Higher activity of ACCase in UV-2 and 5′FDU-1 strongly proves the functional consequences of gene mutation to lipid production. In conclusion, algal mutants exhibited significant impact on biodiesel production through structural alterations in the lipid-metabolizing genes, thereby enhancing lipid production and saturated fatty acid levels.

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Acknowledgments

The authors gratefully acknowledge the Ministry of Earth sciences and Council of Scientific and Industrial Research, Govt. of India, for funding this project. We are also grateful to Dr. R.B.N. Prasad and Dr. Sanjit Kanjilal from Indian Institute of Chemical Technology, Hyderabad for analysing the lipid profile. Dr. Pushpa Viswanathan of Cancer Institute, Chennai and Dr. S. Ramesh of Madras Veterinary College, Chennai are gratefully acknowledged for their help in TEM studies. We would also like to thank Professor T. Subramoniam, D. Sc., F.N.A., for his critical suggestions in this work.

Conflict of Interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Marine Biotechnology Division, ESSO-National Institute of Ocean Technology, Ministry of Earth Sciences, Government of India, Pallikaranai, Chennai, 600 100, India

    Josephine Anthony, Vijaya Raghavan Rangamaran, Dharani Gopal, Kumar T. Shivasankarasubbiah, Mary Leema J. Thilagam, Magesh Peter Dhassiah, Divya Shridhar M. Padinjattayil, VinithKumar N. Valsalan, Vijayakumaran Manambrakat & Kirubagaran Ramalingam

  2. Structural Biology Lab, Department of Bioinformatics, School of Chemical and Biotechnology, SASTRA University, Tanjore, 613 401, India

    Sivakumar Dakshinamurthy & Sivaraman Thirunavukkarasu

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  1. Josephine Anthony
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  2. Vijaya Raghavan Rangamaran
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Correspondence to Josephine Anthony or Kirubagaran Ramalingam.

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Anthony, J., Rangamaran, V.R., Gopal, D. et al. Ultraviolet and 5′Fluorodeoxyuridine Induced Random Mutagenesis in Chlorella vulgaris and Its Impact on Fatty Acid Profile: A New Insight on Lipid-Metabolizing Genes and Structural Characterization of Related Proteins. Mar Biotechnol 17, 66–80 (2015). https://doi.org/10.1007/s10126-014-9597-5

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