Optimization of recombinant maize CDKA production in Escherichia coli: a statistical approach

AAE MÉNDEZ; SM GARZA AGUILAR; LB PENA; SM GALLEGO; JM VÁZQUEZ-RAMOS; SM GALLEGO

Córdoba

Congreso; LII Reunión de SAIB; 2016

SAIB

The complex formed by the cyclin dependent kinase A (CDKA) and cyclin D is responsible for the G1-S transition of cell cycle. Oxidative posttranslational modifications of these proteins block cell cycle progression, and thus contributing in plant growth inhibition during abiotic stress. As part of a project to determine possible sites of CDKA protein oxidation, CDKA from Zea mays L (maize) was cloned and expressed in E. coli. The present study describes the optimization of the production of the protein using as statistical approach: Response Surface Methodology. A 24 full factorial central composite rotary design for four independent variables, each at five levels was employed to fit a second order polynomial model which indicated that 30 experiments were required for this procedure. Level 0 of variables used was 26°C, 0,6 OD 600nm before induction, 0,75 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) concentration and 5h post-induction time. An optimization in CDKA production in the soluble fraction was achieved. Results obtained are not valid for all recombinant proteins.