Project Title: Knowledge-driven improvement of industrial applications of monooxygenase

Supervisor: Dr. Marco van den Berg, DSM Delft, The Netherlands

Objectives: By applied mechanistic studies, the understanding of the kinetics of oxidative enzymes during industrial applications like lignocellulosic biomass hydrolysis or raw material conversion will be improved. This will be achieved by biochemical studies of various available enzymes concerning thermostability, operational stability, kinetic parameters and industrial application. The increased insight will be used to generate knowledge-based enzyme variants, which will be evaluated under relevant conditions.

Methodology: The work will involve various molecular biology and biochemical techniques. Depending on the status of the work, the enzymes to be characterized will be produced either by a bacterial (Escherichia coli) or an eukaryotic (filamentous fungus) host. Enzyme characterization will include: basic in vitro characterization (spectrophotometric assays in MTP), determining enzyme-substrate structure-function relationships, enzyme functioning and mechanisms in application (hydrolysis assays in bio-reactors), stability analyses (thermostability). Some of the activities will be performed in close collaboration with the ESRs in the groups of Prof. Paul Walton (York, UK) and/or Prof. Vincent Eijsink (Ås, Norway).

Expected Results: i). High quality enzyme mutant libraries and selection of improved enzyme variants; (ii). Full characterisation of the tailor-made enzymes by biophysical, kinetic and chemical analyses; (iii). New and/or improved examples of industrial relevant applications of monooxygenases.