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Studies on Squalene Biosynthesis and the Standardization of Its Extraction Methodology from Saccharomyces cerevisiae

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Abstract

In this study, a homogenization-based extraction method was developed and was compared to five conventional methods of squalene extraction. Squalene recovered from this novel procedure gave 3.5-fold, 10-fold, 16-fold, and 8.1-fold higher yield than standard procedures, viz., saponification with 60% KOH, acidic saponification, saponification with 18% KOH, and glass beads method, respectively. Furthermore, this procedure has been evaluated on laboratory Saccharomyces cerevisiae strains such as BY4742 and CEN.PK2-1C (native), deletion strains (ERG6 and ERG11), and tHMG1 overexpressed S. cerevisiae strains. When sonication method of cell lysis was replaced with homogenization, it was found that the yields were significantly higher and reached a value of 9 mg/g DCW in case of BY4742. In addition, squalene yield in ergosterol mutant strains has been analyzed and was found to be 1.8-fold and 3.4-fold higher in ERG6 and ERG11 deletion strains, respectively, than in BY4742. Squalene was also found to be higher at the optimized temperature of 30 °C and pH 6.0. Furthermore, tolerance of S. cerevisiae to external squalene at various concentrations has been carried and found that the organism was tolerant up to 25 g/L of squalene.

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Acknowledgements

KP acknowledges the Department of Biotechnology, India, for the award of research fellowship. SM wishes to acknowledge the financial support provided by the Science Engineering and Research Board (SERB), India. We acknowledge Prof. Ram Rajasekharan (Lipid Science Dept., CSIR-CFTRI) for providing us the strains and the plasmid, pYES2/NTC. The authors are grateful to Mr. K. Anbalagan, CIFS, CFTRI, for his help in SEM analysis and Mr. P. Mukund Lakman, CIFS, CFTRI, for his help during HPLC.

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  1. Microbiology and Fermentation Technology Department, CSIR-Central Food Technological Research Institute, Mysuru, India

    Kalaivani Paramasivan & Sarma Mutturi

  2. Academy of Scientific and Innovative Research, Mysuru, India

    Kalaivani Paramasivan & Sarma Mutturi

  3. University of Madras, Chennai, India

    Kavya Rajagopal

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  1. Kalaivani Paramasivan
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SM and KP designed the experiments, KP and KR performed the experiments, and SM and KP analyzed the data.

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Correspondence to Sarma Mutturi.

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Paramasivan, K., Rajagopal, K. & Mutturi, S. Studies on Squalene Biosynthesis and the Standardization of Its Extraction Methodology from Saccharomyces cerevisiae. Appl Biochem Biotechnol 187, 691–707 (2019). https://doi.org/10.1007/s12010-018-2845-9

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