Bacillus subtilis spore surface display of photodecarboxylase for the transformation of lipids to hydrocarbons

Abstract

The display of enzymes on the surface of spores allows the rapid and very simple biotechnological production of immobilized enzymes. Here we describe the development of a Bacillus subtilis spore display platform and its application to produce hydrocarbons from lipids obtained from the oleaginous yeasts Yarrowia lipolytica and Cutaneotrichosporon oleaginosus as well as olive oil. Lipid hydrolysis was examined in a bienzymatic one-pot cascade using a commercially immobilized lipase as well as spores with and without additional heterologous lipase expression. Decarboxylation of the released fatty acids was achieved displaying a photodecarboxylase from the alga Chlorella variabilis (CvFAP) on the spore surface. Differences in the composition of the formed hydrocarbons were observed to depend on the lipid source. Using 3D printed lighting equipment titers of up to 64.0 ± 5.6 mg L⁻¹ hydrocarbons were produced.