IQUIBICEN   23947
INSTITUTO DE QUIMICA BIOLOGICA DE LA FACULTAD DE CIENCIAS EXACTAS Y NATURALES
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
DEVELOPMENT AND EVALUATION OF NON COVALENT COUPLING METHODS FOR THE PRODUCTION OF ARTIFICIAL CELLULOSOMES
Autor/es:
GOLDBAUM FA; GOROJOVSKY N; CRAIG PO; IGLESIAS RANDO MR
Lugar:
Buenos Aires
Reunión:
Congreso; Reunión Conjunta de Sociedades de Biociencias y XLVI Reunión Anual de la Sociedad Argentina de Biofísica (SAB); 2017
Institución organizadora:
Sociedad Argentina de Biofisica y otras sociedades cientificas SAIB, SAIC, etc
Resumen:
Lignocellulose is the most abundant renewable resource on the planet and it is an excellent substrate for the production of biofuels. Its enzymatic degradation generates sugars that upon fermentation produce bioethanol. For an economically viable production of biofuels it is essential to develop new methods to increase the activity and stability of the enzymes involved in lignocellulose degradation. The cellulosomes of some anaerobic organisms represent the most efficient machinery for the degradation of lignocellulose. These multienzymatic complexes co-localize different cellulolytic enzymes and cellulose binding domains, increasing their degradation activity through enzymatic proximity and substrate targeting effects. Our goal is to develop artificial cellulosomes using an oligomeric protein scaffold that is highly stable and highly expressed in bacteria for the colocalization of cellulases, hemicellulases, beta-glucosidases and cellulose binding domains. For the assembly of these multienzymatic particles we use a non-covalent coupling strategy through high affinity heterodimeric modules complementary fused to the scaffold subunits and the target proteins. In this work we present a comparative analysis of two alternative approaches for non-covalent coupling of enzymes to our oligomeric scaffold using heterodimeric coiled coil peptides and cohesion/dockerin modules. Advantages and disadvantages of each method regarding the expression level, solubility and aggregation tendency of the target and scaffold modules are presented. The analysis of the expression behavior of the isolated protein modules used as building blocks are also presented for comparison. We also show preliminary results on the functional analysis of cellulose binding domains used in the construction of the artificial cellulosomes. It is expected that this technology would be valuable to improve lignocellulose degradation.