High-capacity and Cost-effective Manufacture of Chloroperoxidase

The chloroperoxidase enzyme from the filamentous fungus Caldariomyces fumago has applications in industrial chemical synthesis and the detection and inactivation of chemical warfare agents. Chloroperoxidase is capable of regio- and enantioselective oxygenations and halogenations of organic substrates. When performed chemically, these reactions typically require aggressive reagents and reaction conditions, and lead to the formation of undesired by-products. Widespread adoption of enzyme-catalyzed synthetic strategies is hindered by the high cost of purified proteins, and by the challenges of retaining the native activity of proteins expressed using heterologous host systems. Chloroperoxidase is a heavily glycosylated protein, and only when it is expressed in filamentous fungal hosts such as Aspergillus niger are the post-translational modifications necessary for its activity performed with fidelity. Development of optimized Aspergillus strains and constructs has facilitated heterologous expression of a range of secreted proteins. In this Phase I, Agave BioSystems proposes to develop a system for expression and purification of chloroperoxidase (CPO) from C. fumago using the filamentous fungus Aspergillus niger as host. Fermentation conditions will be optimized for high volume and cost-effective production, and the biochemical properties of the recombinant enzyme will be characterized.