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Purification of Two Isoforms of Pyruvate Kinase from Nordic Krill Using an UNO Q1 Column, Rev A

applied to an UNO Q-1 column (Bio-Rad catalog # 720-0001, see Figure 1). PK I and PK II were well-separated by this procedure at pH 8.0. The previous protocol, using a popular high resolution beaded support, required the use of pH 9.0 buffer to prepare the PK I species, at which pH the enzyme is less stable. The addition of PEP as a substrate stabilizing agent was also required in this pH 9.0 system but not for the UNO-based system. The purified enzyme obtained by this purification process was used to determine that the PK I of Nordic krill does, indeed, exhibit altered physicochemical and kinetic properties with the seasons and may contribute to the temperature adaptations of this species.

Reference
Vetter, R-A.H., and Buchholz, F., Comp. Biochem. Physiol., 116A, 110 (1997).

Data courtesy of Dr. Rolph-Achim Vetter, Biologische Anstalt Helgoland, Meeresstation.


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