Abstract
High concentrations of lipids can obstruct sewer systems and form persistent oil layers. Microbial bioremediation presents a viable solution to mitigate lipid pollution. This study investigates whether the BrkA autotransporter can co-display functional heterodimeric Burkholderia cepacia lipase (LipA) and foldase (LipB) on Escherichia coli UT5600. Surface localization of LipA and LipB was confirmed using trypsin accessibility assays and flow cytometry, indicating lower levels of LipA relative to LipB when individually expressed. Functional analysis was conducted using a para-nitrophenyl palmitate assay and revealed modest lipase activity within co-expressing strains, although activity remained below that of a commercial lipase. To enable unique detection of LipA and LipB within the co-expression system, a Myc-tagged LipB plasmid was generated for use alongside His-tagged LipA. Together, these findings demonstrate the feasibility of BrkA-mediated co-display of functional heterodimeric LipA and LipB. While further optimization is necessary to improve lipase efficiency, these findings provide a foundation for the development of microbial systems for enzymatic lipid bioremediation.