Since inclusion bodies (IBs) contain an important amount of properly folded and active proteins, their solubilization using nondenaturing conditions to obtain aggregation-prone proteins has gained interest. Through these conditions, the refolding step is no longer required, which avoids the usual protein yield loss after this process. Here, we reveal a simple methodology to obtain pure and active difficult-to-produce proteins using two LPS-free expression systems: Lactococcus lactis and Lactobacillus plantarum. This protocol has proven to be successful to obtain proteins which are labile and prone-to-attach (difficult to be purified from other cytoplasmic proteins) and prone-to-aggregate (difficult to be obtained in their soluble form).
Keywords: Inclusion body; Insoluble proteins; L. lactis; Lactobacillus plantarum; Nondenaturing conditions; Solubilization.
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