Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10764651 | Biochemical and Biophysical Research Communications | 2010 | 6 Pages |
Abstract
Here, we report a novel method for large amount production of globular adiponectin from E. coli inclusion bodies utilizing an alkaline-shock solubilization method without chaotropic agents followed by precipitation of the readily renaturing protein. Precipitation of the mildly solubilized protein capitalizes on advantages of inclusion body formation. This approach of inclusion body protein recovery provides access to gram scale amounts of globular adiponectin with standard laboratory equipment avoiding vast dilution or dialysis steps to neutralize the pH and renature the protein, thus saving chemicals and time. The precipitated protein is readily renaturing in buffer, is of adequate purity without a chromatography step and shows biological activity in cultured MCF7 cells and significantly lowered blood glucose levels in mice with streptozotocin induced type 1 diabetes.
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Authors
John T. Heiker, Nora Klöting, Matthias Blüher, Annette G. Beck-Sickinger,