Processing of industrial chemicals by microbial catalysts is a growing and important technology, but at present is more at the research and development stage than at the industrial-commercial one. It is forecasted by Freedonia Group that the catalyst market will grow more into chiral catalyst technologies and in technologies that facilitate new, novel transformations; opportunity where fungal catalyst may thrive. Before biocatalyst could be of industrial relevance, it is necessary to overcome low substrate conversions and low enzymatic expressions, gain control over cell type, and handle mycelia. Beauveria bassiana is a filamentous fungus widely used as a bio pesticide and biocatalyst; it can enhance the yield and conversion of oxidative biotransformations when it is grown in presence of chemical and/or environmental stresses. The purpose of this project is to determine the ability of B.bassiana ATCC 7159 to metabolize the sulfur containing azaarene, phenothiazine, in presence of inducers such as hydrocarbon and insecticides. The underlying hypothesis is that exposure of cells to insecticides and hydrocarbons will increase cells’ oxidative capacity, thus an increase on substrate conversion. Expression of oxidases can increased up to 3 fold when B. bassiana is grown in presence of hexadecane or exposed to 50 ppm insecticides, neonicotinoids and carbamates. Treated and non-treated cells were grown for 7 days in SDB at 28°C and dosed with phenothiazine. Oxidative activity was evaluated via the 7-Ethoxycoumarin O-deethylation assay, a universal oxidase probe. Substrate was measured by GC-MS SIC mode and metabolites characterized by H-NMR. Results include a time course plot, oxidative activity assay, and resting cells assay. Residual phenothiazine was 20% lower in presence of 50 ppm than non-treated cells. The fungus oxidized phenothiazine into phenothiazine sulfoxides, 3-hydroxyphenothiazine sulfoxide, and phenothiazine-3-one. The success of these efforts will provide a new insight into B. bassiana’s oxidative performance.

Methods to increase substrate conversion in the fungus, Beauveria bassiana

249th National Meeting (2015)

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