Production Of Secondary Metabolite By Plant Tissue Culture And Its Downstream Processing

Kasthuri Bai, N. (2006) Production Of Secondary Metabolite By Plant Tissue Culture And Its Downstream Processing. [Student Project Report]

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Abstract

This Dissertation / Report is the outcome of investigation carried out by the creator(s) / author(s) at the department/division of Central Food Technological Research Institute (CFTRI), Mysore mentioned below in this page.

Item Type:	Student Project Report
Additional Information:	An investigation was carried out for the isolation and purification of stevioside from Stevia rebaudiana; and to establish a rapid micropropagation protocol for the production of stevia plants in large number within the time frame. The results are summarized below. Water has been used as an extraction solvent to extract steviosides from the dried leaf powders of Stevia rebaudiana. An easy and quick methodology for identification of the presence of steviosides (qualitatively) using spectrophotometer has been developed to keep in track, that the isolation and purification protocol was efficient and quicker to move from one step to the other. Clarification of the extract using Calcium Oxide followed by Aluminium Oxide removed the pigments efficiently with minimal loss of steviosides when compared to that of Sodium meta bisulfite or Calcium Oxide or Aluminium Oxide alone. The clarified extract obtained after incubation with calcium oxide has been neutralized (before passing through aluminium oxide as mentioned in the previous step) through Carbon-di-oxide bubbling which removes calcium from the extract as Calcium Carbonate. Neutralization through Phosphoric acid was not as efficient as carbon-di-oxide bubbling in the removal of the calcium from the extract. Of various macroporous resins tested for the stevioside adsorption capacity, it was found that IER-NPA I resin adsorbed maximum amount of stevioside from the clarified extract. However, due to their non-availability of required quantity,XAD-2 resin was used for the study. Of various concentrations of alcohol (ethanol and methanol) tested for their desorption capacity for the stevioside from the XAD-2 resin, it was found that 80% ethanol desorbed the steviosides efficiently.Eluted 80% ethanol solution was flash evaporated to concentrate. The concentrate was redissolved in anhydrous methanol and kept overnight in deep freezer (-20oC) for crystallization of stevioside. Such crystallized stevioside was confirmed through Mass Spectrometry. An efficient micropropagation protocol has been developed to produce tissuecultured plants in large number within 50-60 days.
Uncontrolled Keywords:	Secondary Metabolite Plant Tissue Culture Downstream Processing Stevia rebaudiana stevioside micropropagation
Subjects:	600 Technology > 05 Chemical engineering > 01 Biotechnology and Bioengineering 500 Natural Sciences and Mathematics > 10 Plants 500 Natural Sciences and Mathematics > 10 Plants > 05 Tissue Culture
Divisions:	Plant Cell Biotechnology
Depositing User:	Food Sci. & Technol. Information Services
Date Deposited:	22 Dec 2006
Last Modified:	28 Dec 2011 09:26
URI:	http://ir.cftri.res.in/id/eprint/368

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