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.
Similar content being viewed by others
References
Bansal M, Sudhakara Reddy M, Kumar A (2017) Optimization of cell growth and bacoside-A production in suspension cultures of Bacopa monnieri (L.) Wettst. using response surface methodology. In Vitro Cell Dev Biol Plant 53:527–537
Cataldo DA, Maroon M, Schrader LE, Youngs VL (1975) Rapid colorimetric determination of nitrate in plant tissue by nitration of salicylic acid. Commun Soil Sci Plant Anal 6:71–80
Deepak M, Sangli GK, Arun PC, Amit A (2005) Quantitative determination of the major saponin mixture bacoside A in Bacopa monnieri by HPLC. Phytochem Anal 16:24–29
Deepthi S, Satheeshkumar K (2017) Effects of major nutrients, growth regulators and inoculum size on enhanced growth and camptothecin production in adventitious root cultures of Ophiorrhiza mungos L. Biochem Eng J 117:198–209
Eilers RJ, Sullivan JG, Skirvin RM (1988) Analyzing the effects of exogeneous polyamines and growth regulators on plating efficiency of sweet potato protoplasts using a central composite test design. Plant Cell Rep 7:216–219
Gai QY, Jiao J, Luo M, Wang W, Ma W, Zu YG, Fu YJ (2015) Establishment of high-productive Isatis tinctoria L. hairy root cultures: a promising approach for efficient production of bioactive alkaloids. Biochem Eng J 95:37–47
Guillon S, Tremouillaux-Guiller J, Pati PK, Rideau M, Gantet P (2006) Harnessing the potential of hairy roots: dawn of a new era. Trends Biotechnol 24:403–409
Hegazi G, Taha H, Monem Mohamed Sharaf A, Ramzy ES (2017) Enhancing in vitro production of bacoside A from Bacopa monnieriusing precursor and elicitors feeding. J Basic Appl Sci Res 7:27–35
Jadiya P, Khan A, Sammi SR, Kaur S, Mir SS, Nazir A (2011) Anti-Parkinsonian effects of Bacopa monnieri: insights from transgenic and pharmacological Caenorhabditis elegans models of Parkinson's disease. Biochem Biophys Res Commun 413:605–610
Kaur G, Srivastava AK, Chand S (2012) Mathematical modelling approach for concentration and productivity enhancement of 1,3-propanediol using Clostridium diolis. Biochem Eng J 68:34–41
Leonard J, Seth B, Sahu BB, Singh VR, Patra N (2018) Statistical optimization for enhanced bacoside A production in plant cell cultures of Bacopa monnieri. Plant Cell Tissue Organ Cult 133:203–214
Mahato SB, Garai S, Chakravarty AK (2000) Bacopasaponins E and F: two jujubogenin bisdesmosides from Bacopa monniera. Phytochemistry 53:711–714
Majumdar S, Garai S, Jha S (2011) Genetic transformation of Bacopa monnieri by wild type strains of Agrobacterium rhizogenes stimulates production of bacopa saponins in transformed calli and plants. Plant Cell Rep 30:941–954
Monod J (1958) Recherches sur la croissance des cultures bactériennes. Hermann
Murphy J, Riley JP (1962) A modified single solution method for the determination of phosphate in natural waters. Anal Chim Acta 27:31–36
Panda I, Balabantaray S, Sahoo SK, Patra N (2018) Mathematical model of growth and polyhydroxybutyrate production using microbial fermentation of Bacillus subtilis. Chem Eng Commun 205:249–256
Patra N, Srivastava AK (2015) Use of model-based nutrient feeding for improved production of artemisinin by hairy roots of Artemisia Annua in a modified stirred tank bioreactor. Appl Biochem Biotechnol 177:373–388
Patra N, Srivastava AK (2016) Artemisinin production by plant hairy root cultures in gas- and liquid-phase bioreactors. Plant Cell Rep 35:143–153
Patra N, Srivastava AK (2018) Mass production of artemisinin using hairy root cultivation of Artemisia annua in bioreactor. In: Pavlov A, Bley T (eds) Bioprocessing of plant in vitro systems. Springer International Publishing, Cham, pp 343–359
Prakash G, Srivastava AK (2006) Modeling of azadirachtin production by Azadirachta indica and its use for feed forward optimization studies. Biochem Eng J 29:62–68
Prakash G, Srivastava AK (2008) Statistical elicitor optimization studies for the enhancement of azadirachtin production in bioreactor Azadirachta indica cell cultivation. Biochem Eng J 40:218–226
Rahman LU, Verma PC, Singh D, Gupta MM, Banerjee S (2002) Bacoside production by suspension cultures of Bacopa monnieri (L.) Pennell. Biotechnol Lett 24:1427–1429
Ramasamy S, Chin SP, Sukumaran SD, Buckle MJC, Kiew LV, Chung LY (2015) In silico and in vitro analysis of bacoside A aglycones and its derivatives as the constituents responsible for the cognitive effects of Bacopa monnieri. PLoS One 10:e0126565
Ranjan R, Kumar S, Singh AK (2018) An efficient in vitro propagation protocol of local germplasm of Bacopa monnieri (L.) found in Bihar: a plant with wide variety of medicinal properties. J Pharmacogn Phytochem 7:1803–1807
Rastogi S, Pal R, Kulshreshtha DK (1994) Bacoside A3—a triterpenoid saponin from Bacopa monniera. Phytochemistry 36:133–137
Raval KN, Hellwig S, Prakash G, Ramos-Plasencia A, Srivastava A, Buchs J (2003) Necessity of a two-stage process for the production of azadirachtin-related limonoids in suspension cultures of Azadirachta indica. J Biosci Bioeng 96:16–22
Reed J, Osbourn A (2018) Engineering terpenoid production through transient expression in Nicotiana benthamiana. Plant Cell Rep 37:1431–1441
Russo A, Borrelli F (2005) Bacopa monniera, a reputed nootropic plant: an overview. Phytomedicine 12:305–317
Ryu HW, Yuk HJ, An JH, Kim DY, Song HH, Oh SR (2017) Comparison of secondary metabolite changes in Camellia sinensis leaves depending on the growth stage. Food Control 73:916–921
Sharma M, Gupta R, Khajuria RK, Mallubhotla S, Ahuja A (2015) Bacoside biosynthesis during in vitro shoot multiplication in Bacopa monnieri L. Wettst. grown in Growtek and air lift bioreactor. Indian J Biotechnol 14:547–551
Shuler ML, Kargı F (1992) Bioprocess engineering: basic concepts. Prentice Hall, USA
Sivaramakrishna C, Rao CV, Trimurtulu G, Vanisree M, Subbaraju GV (2005) Triterpenoid glycosides from Bacopa monnieri. Phytochemistry 66:2719–2728
Thakore D, Srivastava AK, Sinha AK (2015) Model based fed batch cultivation and elicitation for the overproduction of ajmalicine from hairy roots of Catharanthus roseus. Biochem Eng J 97:73–80
Thakore D, Srivastava AK, Sinha AK (2017) Mass production of ajmalicine by bioreactor cultivation of hairy roots of Catharanthus roseus. Biochem Eng J 119:84–91
Tiwari V, Singh BD, Tiwari KN (1998) Shoot regeneration and somatic embryogenesis from different explants of Brahmi [Bacopa monniera (L.) Wettst.]. Plant Cell Rep 17:538–543
Uabundit N, Wattanathorn J, Mucimapura S, Ingkaninan K (2010) Cognitive enhancement and neuroprotective effects of Bacopa monnieri in Alzheimer's disease model. J Ethnopharmacol 127:26–31
Vijayakumar M, Vijayakumar R, Stephen R (2010) In vitro propagation of Bacopa monnieri L.—a multipurpose medicinal plant. Indian J Sci Technol 3:781–786
Zhong C, Yuan YJ (2009) Responses of Taxus cuspidata to hydrodynamics in bubble column bioreactors with different sparging nozzle sizes. Biochem Eng J 45:100–106
Zote RK, Pati YK, Londhe SS, Thakur VV, Choudhari NB (2018) In vitro regeneration of Bacopa monnieri (L.) from leaf and stem explants. Int J Chem Stud 6:1577–1580
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].
Author information
Authors and Affiliations
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.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s13205-020-02258-6