Investigating The Role of AAL Cassette Proteins in Polysaccharide Biosynthesis in Burkholderia species

Abstract

It is hypothesized that the Amino Acid Ligase (AAL) - Acyl Carrier Protein (ACP) complex, encoded within the AAL cassette in Burkholderia, is essential for the first stage of surface exopolysaccharide (EPS) production, which is a crucial component in the formation of bacterial biofilms and offers insights into their roles in both antibiotic resistance and biofilm formation. AAL: ACP1 and ACP2 proteins were produced separately in E. coli, purified, and modified using an ideal buffer system to test this theory. Subsequently, to verify modification and evaluate protein-protein interactions, the modified proteins were examined using native gel electrophoresis, mass spectrometry, and DTNB tests. In addition, the amino acid transfer activity was assessed using the AMP-Glo assay. ACP1 was successfully modified by the study, whereas only minor alterations were observed in ACP2. However, using native gel electrophoresis, no persistent interactions between AAL and the ACPs were found, and the AMP-Glo assay showed difficulties in identifying amino acid transfer, indicating the need for additional adjustments. To further comprehend AAL-ACP interactions, future research should concentrate on improving the modification processes and using more sensitive approaches. These developments could help create fresh tactics to combat Burkholderia species' biofilm growth and antibiotic resistance.