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Proper protein function is essential to health, and protein mismanagement often leads to disease, so efficient protein characterization is critical. There are many modern tools available to characterize an unknown protein. The BASIL system (Biochemistry Authentic Scientific Inquiry Lab) provides a structured methodology to this end. This study uses BASIL to characterize the protein 2O14. BL21(DE3) competent cells were transformed with a vector including the lac operon and then incubated in the absence of glucose to induce 2O14 production. The protein was purified from cellular lysate using affinity column chromatography, and molecular weight was determined to be 42 kDa via SDS-PAGE. Purified protein concentration was found to be around 8 mg/mL spectrophotometrically, indicating efficient purification. In silico approaches through BLAST and Pfam suggest lipase activity for 2O14. An enzyme activity assay was conducted to measure lipase activity spectrophotometrically, and found it was highly active for the predicted lipase function.
Belville, AJ and Vogelei, Andrew, "Purification and Characterization of the Protein 2O14" (2021). Biochemistry and Molecular Biology Presentations. 22.
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