Use of BugBuster® and Lysonase™ Reagents for efficient protein extraction from Gram-positive bacteria
BugBuster® Protein Extraction Reagent has become a valuable tool for extraction of proteins from E. coli without the need for mechanical disruption. It is also known that supplementing BugBuster® (as well as its derivative, PopCulture™ Reagent) with a small amount of lysozyme enhances the efficiency of extraction from E. coli, especially for larger proteins1. Furthermore, the addition of Benzonase® Nuclease to the extraction mixture degrades the liberated nucleic acids, reducing viscosity and potential interference of DNA and/or RNA with purification procedures2. With this issue of inNovations, we introduce Lysonase™ Bioprocessing Reagent, which is an optimized blend of rLysozyme™ Solution and Benzonase® Nuclease that can be used alone or in conjunction with BugBuster® or PopCulture™ Reagent for any of their applications.
Figure 1 shows that the combination of BugBuster® and Lysonase™ greatly enhanced the release of soluble proteins from two different species of Gram-positive bacteria. Extraction is accomplished by resuspending cells in BugBuster® containing 1/125 volume of Lysonase™ followed by a 20-minute incubation at room temperature. Insoluble material is simply removed by centrifugation and the supernatant can be used directly for purification or activity assays.
Figure 1. Combination of BugBuster® and Lysonase™ greatly enhanced the release of soluble proteins from two different species of Gram-positive bacteria. Extraction is accomplished by resuspending cells in BugBuster® containing 1/125 volume of Lysonase™ followed by a 20-minute incubation at room temperature. Insoluble material is simply removed by centrifugation and the supernatant can be used directly for purification or activity assays.
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