IMPROVING LABORATORY TOOLS TO STUDY STARCH EXCESS 4 (SEX4) ALGAE HOMOLOGOUS ENZYMES

CARRILLO, J.; MARTIN, M; GOMEZ-CASATI, D.F; BUSI, M.V.

Rosario

Congreso; L Reunión Anual Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2014

SAIB - Sociedad Argentina de Investigación en Bioquímica y Biología Molecular

Phosphoglucan phosphatases are novel enzymes that remove phosphates from complex carbohydrates. In plants, these proteins have a regulatory role in the night time remobilization of leaf starch being SEX4, which primary structure contains a DSP (dual-specificity phosphatase) as well as a CBM (carbohydrate-binding module) domain, its major representative member. To understand the evolution of catalysis and regulation of these enzymes we decided to investigate SEX4 homologous enzymes in eukaryotic green algae. These organisms are an excellent model toinvestigate the properties of photosynthetic physiology because they present the critical photosynthetic functions as higher plants but a decreased gene redundancy. To this end we first set up laboratory conditions to culture and cryopreservate Ostreococcus tauri, Nannochloropsis gaditana, Micromonas pusilla and Chlamydomonas reinhardtii strains. We tested and optimized different total RNA, DNA and protein extraction procedures in order to find the bestisolation protocol for each macromolecule in terms of quality and yield. These laboratory tools will permit us to analyze evolutionary aspects related to phosphoglucan phosphatases from O. tauri, the smallest free-living eukaryoteidentified to date.