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The strains were propagated in RBC (A+) in complete medium (RPMI 1640 medium supplemented with 0.5% Albumax, 9 mM glucose, 0.2 mM hypoxanthine, 2.1 mM L-glutamine, and 22 g/ml gentamycin) at 3.3% hematocrit and 37C in a gaseous mixture consisting of 3% O 2 , 3% CO 2 , and 94% N 2

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However, several factors, such as non-protein compounds, interfere with the hydrolysis process, so the initial extraction of crude protein is crucial

J Biol Chem 248(1):2532 Busiello RA, Savarese S, Lombardi A (2015) Mitochondrial uncoupling proteins and energy metabolism