SBIR-STTR Award

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.

Keywords:
Chloroperoxidase, Aspergillus, Decontamination, Fermentation, Heterologous Protein Expression, Chemical Warfare Agent, Recombinant Enzyme

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. In the Phase I, Agave BioSystems and collaborators from Cornell University achieved high level production of highly active recombinant Caldariomyces fumago chloroperoxidase (rCPO) in Aspergillus. The current not fully optimized levels of expression are five-fold higher than previously reported. They are also 100 fold lower than potential levels which have been achieved with other Aspergillus systems providing enormous upside potential that will be realized upon completion of the Phase II tasks. The result will be an expression system that meets or exceeds the sponsor?s specifications.

Keywords:
Chloroperoxidase, Aspergillus, Decontamination, Fermentation, Heterologous Protein Expression, Chemi