@inproceedings{8763632,
  abstract     = {{Designer cellulosomes are engineered multi-enzyme complexes, comprising both cellulases and hemicellulases attached to a common backbone, the scaffoldin. The colocalization of complementary enzymatic activities causes a large intermolecular synergistic effect, improving degradation efficiencies. Therefore, these complexes can play a significant role in efficient hydrolysis of lignocellulosic biomass.
Despite these superior opportunities of designer cellulosomes compared to free (hemi)cellulases, they have not yet come to commercial use. This problem can be contributed to two main reasons. (i) Designer cellulosomes comprise multiple modules. These modules need to be individually assembled on molecular level, followed by tedious procedures for recombinant expression, purification, and in vitro construction of the complex. In addition, previous work has shown that many parameters, e.g. module combinations and position, have an influence on the efficiency of designer cellulosomes. (ii) Lignocellulosic biomass is a complex and variable substrate. The total saccharification of plant cell walls thus requires a vast mixture of enzymes with various activities and specificities. Both aspects contribute to a multi-parametrical, labour intensive and expensive optimization process for each designer cellulosome. This is prohibitive for rapid adaptation of the cellulosome composition towards the available lignocellulose substrate.
The goal of this research is to overcome these hurdles by creating a cellulosome library, followed by a suitable selection process. First, we are able to do rapid and convenient construction of designer cellulosomes on molecular level, using the VersaTile technique. This is an in-house developed technique dedicated to the assembly of modular proteins by ‘clicking’ molecular building blocks at will. Our currently available repository comprises 142 building blocks encoding different CAZymes, targeting cellulose and all major hemicelluloses, in addition to dockerins, cohesins and CBMs.
Secondly, the difficulties of in vitro assembly of cellulosomes can be solved by yeast cell surface display. When switching to an in vivo system, complexes are produced by the yeast cell itself, exempting protein engineers from much work and costs. An automatic assembly allows us to fully exploit the combinatorial power of VersaTile. By performing random shuffling of all the available building blocks, a library of theoretically 4x1018 different cellulosomes can be constructed. A selection process will result in an arsenal of the most optimal complexes for the saccharification of a certain type of lignocellulosic substrate, contributing to the industrial competitiveness of designer cellulosomes.}},
  author       = {{Lamote, Babette and Cremelie, Emma and Vanderstraeten, Julie and de Carvalho Maurício da Fonseca, Maria João and De Mey, Marjan and Briers, Yves}},
  booktitle    = {{International Conference on Renewable Resources & Biorefineries, 18th, Abstracts}},
  language     = {{eng}},
  location     = {{Bruges, Belgium}},
  title        = {{Towards industrial competitiveness of designer cellulosomes using the VersaTile technique}},
  year         = {{2022}},
}