Scientific approaches and hypotheses

The conditions of industrial saccharification vary markedly from those typically found during enzymatic break down of cellulose in Nature. Therefore, “natural enzymes” (wild-types) may not be optimal for the industrial process. In RESAB we use rational and semi-rational approaches to find enzyme variants with faster catalysis under industrial conditions. The work is based on iterative circles which may be coarsely described as:

  1. Identification of potential bottlenecks
    We characterize the kinetics of a given cellulase experimentally and model the experimental data to elucidate molecular origins of the rate limiting steps in the catalytic cycle.
  2. Design of variants
    We analyze the structure of the wild-type enzyme and devise mutations that are likely to make the enzyme less prone to the rate limiting factor identified under 1.
  3. Produce variants
    We use site directed mutagenesis to express mutant enzymes. These enzyme variants are then shuttled into point 1 for update of hypothesis and continuous optimization.