Affinity-based isolation of a bacterial lipase through steric chaperone interactions
| Autor: | Kris Pauwels, Patrick Van Gelder |
|---|---|
| Přispěvatelé: | Structural Biology Brussels, Department of Bio-engineering Sciences |
| Rok vydání: | 2008 |
| Předmět: |
Lipopolysaccharides
Circular dichroism biology Burkholderia lipopolysaccharide affinity purification Lipase biology.organism_classification Chromatography Affinity Bacterial Proteins Biochemistry Affinity chromatography protein purification Fermentation Foldase Protein purification biology.protein Burkholderia glumae Denaturation (biochemistry) Molecular Chaperones Biotechnology Thermostability |
| Zdroj: | Protein Expression and Purification. 59:342-348 |
| ISSN: | 1046-5928 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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