The production and consumption of molecular hydrogen drives the physiology and bioenergetics of many microorganisms in hydrothermal environments. As such, the potential of these microorganisms as model systems to probe fundamental issues related to biohydrogen production merits consideration. Specifically, it is important to understand how carbon/energy sources relate to the disposition of reducing power and, ultimately, the formation of molecular hydrogen by high temperature microorganisms. This project will focus on bacteria of the thermophilic order Thermotogales, fermentative anaerobes that produce H2 from simple and complex carbohydrates. A combination of metabolomics, genomics and genetics will be used to interrogate the Thermotoga system to expand understanding of biohydrogenesis.

The limited number of therapeutic exotoxins approved by the FDA suffer from structural limitations, drastically reducing their therapeutic value. This problem arises from an inability to produce versions of these proteins exhibiting humanized glycosylation patterns, resulting in a short molecular half-life and insufficient interaction with the target site during treatment. Production of glycosylated exotoxins is difficult because conventional production platforms designed to produce glycosylated proteins are susceptible to the toxicity of the exotoxins themselves. To overcome these limitations, cell lines were developed that are exotoxin resistant and produce glycosylated versions of these proteins. Also, cell lines were programmed to secrete the potentially therapeutic exotoxins into the growth media. These results will stimulate adoption of natively glycosylated therapeutic proteins across cancer treatment and other diseases.

Many biological toxins are known to attack specific cell types, delivering their enzymatic payloads to the cytosol. This process can be manipulated by molecular engineering of chimeric toxins. The Clostridium botulinum C2 binding/translocation domain was retargeted to neural cell populations by deleting its non-specific binding domain and replacing it with a C. botulinum neurotoxin binding domain. This retargeted toxin may enable delivery of therapeutics to peripheral neurons and be of use in addressing experimental questions about neural physiology.