Recovery And Evaluation Of Endoglucanase And Xylanase Enzymes In Agroinfiltrated Nicotiana Benthamiana Leaves | 12221
Journal of Biotechnology & Biomaterials
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An economical method for the conversion of renewable biomass (cellulose and hemicellulose) to monosaccharide
components, which can be fermented to produce biofuels through saccharification, is to use plants as bioreactors for
cellulases production. The present work, was focused in the recovery and analysis of Endo-?-D-xylanase (EC 126.96.36.199), both
individually and in co-expression with Endo-?-1, 4-glucanase (EC 188.8.131.52) in the same Nicotiana benthamiana tissue. The genes
of interest were delivered into the leaves by vacuum agroinfiltration.
We evaluated in detached N. benthamina leaves, the expression of xylanase, using two kinds of plasmids; the first acts under
the control of the constitutive CaMV 35S promoter (pDX:xyl), and the second is a virus-based expression system (CMVva:xyl).
We also performed co-infiltration of xylanase with 3 differente gene silencing supressors (GSS): p1, p19 and p25. As a result, at
4 days post-infiltration, the xylanase expression was higher when coinfiltration with p19 and using the pDX:xyl yielding up to
17.22mg/kg FW and 0.3% of the TSP. The co-expression xylanase-endoglucanase in the same tissue was possible, with a level of
16.3mg/kg FW (0.2%TSP) and 17.18mg/kg FW (0.21% TSP) respectively, when performed in coinfiltration with p19.
The activity of the xylanase under different storage conditions (RT, 4?C, -20?C and -80?C) was estimated, after 0, 7 and 21
days. Our results indicate that the activity of the xylanase is stable for at least 21days, preserving more that 88% of the activity at
RT and -80?C storage, and 80% at 4?C and -20?C.
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