Optimal expression of GUS gene from methyl jasmonate-inducible promoter in high density culture of transformed tobacco cell line BY-2

Bioreactor Engineering For Recombinant Protein Production Using Plant Cell Suspension Culture

“…Cells may continue to produce the recombinant protein upon initial entering into the stationary phase of the growth cycle, but this is usually accompanied with increased proteolytic activities, and hence the recombinant protein level tends to descend during the stationary phase when the 35S promoter is used. If an inducible promoter is used, generally the transgene is induced after the culture reaches a high biomass concentration in the late/post exponential growth phase [59]. In this case, recombinant protein production is decoupled from the active cell growth.…”

“…A number of inducible promoters have been used for expressing recombinant proteins in plant suspension cultures. The rice -amylase (RAmy3D) promoter which is induced by sugar starvation was used in rice cell cultures to express recombinant 1 -antitrypsin [15,16] and recombinant hGM-CSF [5]; the Arabidopsis thaliana heat-shock (HSP18.2) promoter [60], the tomato light inducible rbcS promoter [61], the methyl jasmonate inducible potato cathepsin D inhibitor (CDI) promoter [59], the glucocorticoid-inducible GVG promoter [62], the sweet potato oxidative stress-inducible peroxidase (POD) promoter [63], and the abscisic acid, tetracycline, and copper inducible promoters [64], have all been examined in tobacco cell cultures for recombinant protein production. In order to optimize the efficiency of an inducible gene expression system, it is necessary to examine the inducer concentration and timing of inducer addition.…”

“…Published data in this area for plant cell culture is scarce. Suehara et al [59] investigated optimal induction strategies for the expression of a GUS reporter driven by the CDI promoter. Since the addition of the inducer, methyl jasmonate, led to metabolic by-products that reduced cell growth, Suehara et al [59] had to replace the spent media with fresh ones to remove the potential inhibitory substances, and to devise an inducer feeding strategy by keeping the inducer concentration within a narrow range.…”

“…Suehara et al [59] investigated optimal induction strategies for the expression of a GUS reporter driven by the CDI promoter. Since the addition of the inducer, methyl jasmonate, led to metabolic by-products that reduced cell growth, Suehara et al [59] had to replace the spent media with fresh ones to remove the potential inhibitory substances, and to devise an inducer feeding strategy by keeping the inducer concentration within a narrow range. Trexler et al [16] postulated that expression systems based on the rice -amylase promoter could be further improved by optimizing the timing of medium exchange using suitable physiological indicators, and by exposing the culture to consecutive growth and sugar-starvation phases.…”

“…While it is most common to culture plant cells in the single-stage batch mode, two-stage batch [15], fed-batch [23,59], chemostat [75], and perfusion modes [34,76] have also been explored for protein production from cultured plant cells [4]. As discussed in the previous section, choice of bioreactor operation mode is largely governed by the pattern of product formation and the way the product is translocated following its synthesis.…”