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Cost Effective Approach for Production of Chlorella pyrenoidosa: A RSM Based Study

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Microalgae are considered as a promising feedstock for biodiesel production due to their faster growth rate, higher biomass productivity and lipid content which ranges from 20 to 50% of its dry weight. In this work, optimization of bold basal media (BBM) for the production of Chlorella pyrenoidosa are studied with central composite design (CCD) using response surface methodology. The classical optimization is used for identifying the best pH for the growth of C. pyrenoidosa and found to be 8. The screening of significant factors for the growth was studied using Plackett–Burman design (PBD) with 7 factor experiment. The significant factors identified were MgSO4⋅7H2O, CaCl2⋅2H2O, KNO3, Na2EDTA from PBD of experiment. These significant factors were included in a CCD, resulting in 31 experiments, that were further performed. The chlorophyll a, chlorophyll b, chlorophyll a, b and turbidity were measured regularly during experiments. The optimum concentration of MgSO4⋅7H2O, CaCl2⋅2H2O, KNO3, and Na2EDTA were found to be 0.1, 0.0375, 0.375, and 0.0625 g/L, respectively. The turbidity, chlorophyll a, chlorophyll b and chlorophyll a, b obtained with optimum concentrations of BBM were 0.809, 5.7504, 1.6137 and 7.3642 mg/L respectively. The results showed a yield of more than 5 times higher with optimized BBM as compared to the actual media. Moreover, the effect of light and urea on C. pyrenoidosa growth were studied and it was found that turbidity, chlorophyll a, chlorophyll b and chlorophyll a, b were maximum when an artificial light of 3500 lx was used. The nitrogen starvation during the growth of C. pyrenoidoa showed a maximum lipid content of 36.03% dry weight on 16th day which is quite sufficient for the production of biodiesel. Hence, C. pyrenoidosa could be cultivated with the optimum BBM for biodiesel production.

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Fig. 8


A665 :

Absorbance at 665 nm

A650 :

Absorbance at 650 nm

OD680 :

Optical density at 680 nm

Y1 :

Turbidity at 680 nm

Y2 :

Chlorophyll a content in mg/L

Y3 :

Chlorophyll b content in mg/L

Y4 :

Chlorophyll a, b content in mg/L


Degrees of freedom


Bold basal media


Analysis of variance


Response surface methodology


Central composite design


Plackett–Burman design


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The authors would like to thank Mechanical Engineering Department and School of Biotechnology, National Institute of Technology Calicut, and India for providing necessary lab facility for doing this work. The authors are thankful to Kerala State Council for Science, Technology and Environment (KSCSTE), Kerala, India for financially supporting (Order No. 1237/2015/KSCSTE) this investigation. A funding agency was involved in this work and facilities given by institute were acknowledged separately. This manuscript has not submitted elsewhere for the consideration of other journals and the manuscript has not been published partly/ fully. The results and conclusions given in the manuscript are original. No data, text, or theories by others are presented without mentioning the reference and as if they were the author’s own (“plagiarism”).

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Correspondence to A. Shaija.

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Mubarak, M., Shaija, A. & Suchithra, T.V. Cost Effective Approach for Production of Chlorella pyrenoidosa: A RSM Based Study. Waste Biomass Valor 10, 3307–3319 (2019).

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