Enabling Ethanologenesis in Moorella thermoacetica through Construction of a Replicating Shuttle Vector

Replicating plasmid shuttle vectors are key tools for efficient genetic and metabolic engineering applications, allowing the development of sustainable bioprocesses using non-model organisms with unique capabilities. To date, very limited manipulation has been achieved in thermophilic acetogen, Moorella thermoacetica, partly due to lack suitable vectors. However, M. thermoacetica considerable potential as an industrial chassis organism, which can only be unlocked if reliable effective place. This study reports construction a replicating vector through identification implementation compatible Gram-positive replicon allow maintenance within host. Although characterisation behaviour proved difficult, designed was subsequently applied ethanologenesis, i.e., ethanol production this organism. The non-native ethanologenesis via plasmid-borne overexpression native aldh gene heterologous expression Clostridium autoethanogenum adhE1 gene. result demonstrates importance developed efforts industrially important thermoacetica.