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; IGLESIAS RANDO MR; GOROJOVSKY N; CRAIG PO
Lugar:
Buenos Aires
Reunión:
Simposio; Simposio Fronteras en Biociencias 3; 2018
Resumen:
Lignocellulose is the most abundant renewable resource on the planet and it is an excellent substrate for the production of biofuels. Its enzymatic degradationgenerates sugars that upon fermentation produce bioethanol. For an economicallyviable 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 throughhigh affinity heterodimeric modules complementary fused to the scaffold subunitsand the target proteins. In this work we present a comparative analysis of twoalternative approaches for non-covalent coupling of enzymes to our oligomericscaffold using heterodimeric coiled coil peptides and cohesin/dockerin modules.Advantages and disadvantages of each method regarding the expression level,solubility and aggregation tendency of the target and scaffold modules arepresented. The analysis of the expression behavior of the isolated protein modules used as building blocks is also presented for comparison. We also show results for the functional analysis of cellulose binding domains and for some enzymes coupled to the scaffold as well as on their own. It is expected that this technology would be valuable to improve lignocellulose degradation.