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Statistical optimization of bacoside A biosynthesis in plant cell suspension cultures using response surface methodology

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Abstract

The in vitro cultures of Bacopa monnieri show poor production of the anti-Alzheimer’s drug, bacoside A. Therefore, suitable bioprocess optimization strategy was developed for callus induction from leaf explants (30 days), followed by callus proliferation (15 days). Central Composite Design was implemented to analyze the effect of pH, photoperiod, naphthalene acetic acid (NAA), and benzylaminopurine (BAP) concentration for maximum biosynthesis of bacoside A using leaf explants as well as callus explants as the inoculum. Using the CCD responses, it was predicted that the best biomass concentration of 4.56 ± 0.53 g/l DW and bacoside A production of 14.04 ± 1.31 mg/g DW can be obtained using 5.4 pH, 18 h/6 h L/D photoperiod regime, and 1.2 mg/l BAP in combination with 0.2 mg/l NAA. The kinetic parameter values for maximum specific growth rate (0.16/day), saturation constant (7.35 g/l), inhibition constant (120 g/l), biomass yield (0.011 g/g), maintenance coefficient (0.02 g/g/day), and growth-associated (0.627 mg/g) and non-growth-associated (1.096 mg/g/day) bacoside A formation constants were determined experimentally in batch cultures using optimized conditions.

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Acknowledgements

The authors obtained the elite plant material from CIMAP Lucknow. The financial support was provided by SERB (File number: ECR/2017/001113) (Govt. of India) to one of the authors for pursuing PhD (Bishwanath Seth).MHRD (India) provided the fellowship for pursuing M.Tech. to two of the authors (Krishna Kalyani Sahoo and K.R. Aravind). The authors are also thankful to the Life Science Department of NIT Rourkela for providing the HPLC facility which was funded by DST (FIST), India [File number: SR/FST/LSI-025/2014].

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Authors and Affiliations

  1. Department of Biotechnology and Medical Engineering, National Institute of Technology (NIT), Rourkela, 769008, India

    Bishwanath Seth, Krishna Kalyani Sahoo, K. R. Aravind & Nivedita Patra

  2. Department of Life Science, NIT, Rourkela, 769008, India

    Binod B. Sahu

  3. Central Institute of Medicinal and Aromatic Plants (CIMAP), P.O. CIMAP, Lucknow, 226016, India

    V. R. Singh

Authors
  1. Bishwanath Seth
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  2. Krishna Kalyani Sahoo
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  3. K. R. Aravind
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  4. Binod B. Sahu
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  5. V. R. Singh
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  6. Nivedita Patra
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Contributions

The kinetic studies, and spectrophotometric and HPLC analysis have been standardized and performed by Bishwanath Seth. Krishna Kalyani Sahoo performed the RSM experiments for phytohormones, its experimental validation, and substrate inhibition studies. K. R. Aravind performed RSM experiments for pH and photoperiod optimization. B. B. Sahu and V. R. Singh were the collaborators for this work. Nivedita Patra has been the thesis supervisor for Bishwanath Seth, Krishna Kalyani Sahoo, and Aravind in this research work.

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Correspondence to Nivedita Patra.

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Seth, B., Sahoo, K.K., Aravind, K.R. et al. Statistical optimization of bacoside A biosynthesis in plant cell suspension cultures using response surface methodology. 3 Biotech 10, 264 (2020). https://doi.org/10.1007/s13205-020-02258-6

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