title: Affinity-based isolation of a bacterial lipase through steric chaperone interactions.
abstract: Lipases are important as additives in detergent formulations but their biocatalytic potential is increasingly exploited in the synthesis of high-added value chemicals, in fine-chemical production and in the pharmaceutical industry. Traditionally, conventional purification schemes comprise several chromatographic steps. Here we report a new purification procedure of the lipase (LipA) that is endogenously secreted by the Gram-negative bacterium Burkholderia glumae. This affinity purification combines the specific binding scaffold of a lipase-specific foldase (Lif) and the intrinsic resistance to chemical denaturation of LipA. The newly devised method is less labor-intensive, is fast, leads to a homogeneous preparation and can be easily scaled up. The novel and the conventional purification strategies were evaluated in parallel and characteristics of the B. glumae lipase were analyzed via CD spectroscopy. Lipopolysaccharide (LPS) was still present in the samples purified via the conventional purification scheme and was shown to increase the thermostability of the lipase.
authors.affiliation: Department of Molecular and Cellular Interactions, VIB and Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium. krpauwel@vub.ac.be
authors.forename: Kris
authors.initials: K
authors.lastname: Pauwels
authors.forename: Patrick
authors.initials: P
authors.lastname: Van Gelder
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification*; D019117:Burkholderia / Q000201:enzymology*; D002846:Chromatography, Affinity / Q000379:methods*; D005285:Fermentation; D008049:Lipase / Q000096:biosynthesis / Q000737:chemistry*; D008070:Lipopolysaccharides / Q000737:chemistry; D018832:Molecular Chaperones / Q000096:biosynthesis / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; C079999:LipA protein, Bacteria; D008070:Lipopolysaccharides; D018832:Molecular Chaperones; D008049:Lipase; C504169:lipase foldase