BugBuster® Plus Lysonase™ Kit: Cell lysis and nucleic acid removal for Gram-negative and Gram-positive bacteria

BugBuster® Protein Extraction Reagent has become an indispensable tool for efficient extraction of active soluble proteins from E. coli without the need for sonication or any other method of mechanical disruption. Lysonase™ Bioprocessing Reagent is a blend of rLysozyme™, a highly purified and stabilized recombinant lysozyme, and Benzonase® Nuclease, a recombinant nonspecific endonuclease that degrades all forms of DNA and RNA. The new BugBuster Plus Lysonase Kit combines the activities of both reagents to significantly increase protein extraction efficiency and facilitate downstream processing of protein extracts, thereby enabling maximum recovery of active soluble protein from both Gram-negative and Gram-positive bacteria. Use 5 ml BugBuster and 10 μl Lysonase per gram of cell paste. Two kit sizes provides sufficient reagents for protein extraction from either 20 or 100 g cell paste.

Protein extraction from Gram-positive bacteria.

Figure 1. Protein extraction from Gram-positive bacteria. Cell pellets (25 mg) of Bacillus subtilis and Enterococcus faecalis were resuspended in 250 μl BugBuster containing 2 μl Lysonase Bioprocessing Reagent or in 250 μl 50 mM Tris-HCl (pH 8.0) containing 2 μl Lysonase. Samples were incubated for 10 min at room temperature and cell debris was pelleted by centrifugation for 10 min at 14,000 g. Samples (15 μl) of the supernatants were analyzed by SDS-PAGE (10–20% gradient gel) and Coomassie blue staining.

Materials