CKB Human, Pichia

CKBB is a cytoplasmic enzyme that typically functions as a homodimer in the brain and other tissues. It can also form heterodimers with muscle isoforms in the heart. The enzyme’s primary function is to catalyze the conversion of creatine and ATP to phosphocreatine and ADP, a reaction that is vital for maintaining cellular energy balance .

The recombinant form of CKBB is produced using the yeast Pichia pastoris. This method involves inserting the gene encoding CKBB into the Pichia pastoris genome, allowing the yeast to express the enzyme. The recombinant CKBB produced in this way is glycosylated and has an amino acid sequence identical to the native enzyme .

The recombinant CKBB is purified under non-denaturing conditions, ensuring that it retains its native structure and function. It is typically purified using proprietary chromatographic techniques. The enzyme is stored below -18°C to maintain its stability and prevent freeze-thaw cycles, which can degrade its activity .

Recombinant CKBB is used in various research applications, including studies on energy metabolism, enzyme kinetics, and cellular physiology. It is also used in diagnostic assays to measure creatine kinase activity in biological samples .

The biological activity of recombinant CKBB is measured by its enzymatic activity, specifically its ability to catalyze the conversion of creatine phosphate. The specific activity of the enzyme is typically around 854 IU/mg at 37°C, corresponding to a specific activity of 1,171 ng/ml .

The recombinant CKBB is formulated in a solution containing 10 mM Bis-Tris-HCl (pH 6.0), 50% glycerol, 0.5 mM EDTA, and 0.5 mM DTT. This formulation helps to maintain the enzyme’s stability and activity during storage and use .