Regresar

Production of grapevine cell biomass (Vitis Vinifera L. Gamay) and resveratrol in custom commercial bioreactors using cyclodextrins methyl-jasmonate elicitors

Abstract:

Plant cell liquid culture may become an alternative to produce valuable secondary metabolites instead of extracting from plant raw material. Besides the optimization of the culture medium and culture conditions of highly productive cell lines, the culture scale-up in a suitable bioreactor is a compulsory step for bioprocess development. We have designed an airlift-type bioreactor (ALB) of low cost featured for aseptically whole medium extraction and replacement. Medium exchange enables the modification of the culture conditions and the recovery of metabolites secreted to the medium as advantageous features. The suitability of ALB for plant cell culture has been tested by comparing its performance with the commercial stirred and aerated tank bioreactor (SATB)-type Biostat B to grow up grapevine cells. The comparison has been realized under a randomized block experimental design, taking the bioreactor type and the sucrose concentration as factors. Biomass yield (g biomass. substrate-1), productivity (g biomass. g substrate-1.day-1), maximum biomass production, final biomass concentration, maximum growth rate, sucrose consumption rate and glucose usage were determined at three different levels of sucrose concentration, low, medium and high (10, 20, 30 g.L-1 respectively) in both bioreactors in two replicates. The cell growth rate during the exponential phase (μ) was not significantly affected by either the bioreactor type or the sucrose concentration, but in the SATB the exponential phase extended for longer times than in ALB, thus leading to obtain more final biomass and higher productivity variables. Lower oxygen availability could be a causal factor for the less efficient use of the carbon source in ALB, but this limitation might be solved by a redesign of an improved ALB prototype. To achieve extracellular resveratrol production, culture media were supplemented with elicitors and incubated with cells in the bioreactors for several days. Upon elicitation with methylated cyclodextrin, the medium in ALB reached a final concentration of 0.58 mg.mL-1 and a specific production of 2.4 mg. g biomass-1 while in STAB was 1.01 mg.mL-1 and 3.1 mg. g biomass-1 respectively. When methylated cyclodextrin were used in combination with methyl jasmonate the resveratrol production largely increased and was similar in both bioreactors, final concentration above 4.50 mg.mL-1 and specific production above 13.0 mg. g biomass-1. Although the growth of grapevine cells in ALB was not as effective as in STAB, the design of ALB bioreactor showed a great potential for the production of resveratrol. © 2013 by Nova Science Publishers, Inc. All rights reserved.