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Optimization and elucidation of interactions between ammonium, nitrate and phosphate inCentella asiatica cell culture using response surface methodology
Authors:Rozita Omar  M A Abdullah  M A Hasan  M Marziah  M K Siti Mazlina
Institution:(1) Department of Chemical and Environmental Engineering, Faculty of Engineering, Universiti Putra Malaysia, Perak, Malaysia;(2) Department of Bioprocess Technology, Faculty of Biotechnology and Molecular Science, Universiti Putra Malaysia, Perak, Malaysia;(3) Department of Chemical Engineering, Universiti Teknologi Petronas, Perak, Malaysia;(4) Department of Biochemistry, Faculty of Biotechnology and Molecular Science, Universiti Putra Malaysia, Selangor, Malaysia
Abstract:The effects of macronutrients (NO3 , NH4 + and PO4 3−) on cell growth and triterpenoids production inCentella asiatica cell suspension cultures were analyzed using the Box-Behnken response surface model experimental design. In screening and optimization experiments, PO4 3− as a single factor significantly influenced cell growth where increasing the phosphate level from 0.1 to 2.4 or 2.6 mM, elevated cell growth from 3.9 to 14–16 g/L. The optimum values predicted from the response surface model are 5.05 mM NH4 +, 15.0 mM NO3 and 2.6 mM PO4 3−, yielding 16.0 g/L cell dry weight with 99% fitness to the experimental data. While the NH4 +-NO3 interaction influenced cell growth positively in the optimization experiment, NH4 + and NO3 as single factors; and interactions of NO3 -PO4 3−, NH4 +-PO4 3− and NH4 +-NO3 were all negative in the screening experiment. Cell growth and the final pH level were positively affected by PO4 3−, but negatively affected by NH4 + and NH4 +-PO4 3− interactions. The different effects of factors and their interactions on cell growth and final pH are influenced by a broad or narrow range of macronutrient concentrations. The productions of triterpenoids however were lower than 4 mg/g cell dry weight.
Keywords:response surface methodology            Centella asiatica            optimization  plant cell culture  modelling  fermentation
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