Overproduction of a recombinant heat-stable catalase for use in an extremophile enzyme lab exercise

The manganese-dependent catalase of <i>Pyrobaculum calidifontis</i>, a facultative aerobe that grows optimally at 90 ˚C, has been previously used in a teaching lab to highlight salient characteristics of hyperthermophilic extremophiles. However, accruing enough enzyme from the native organism, which would allow each student in a typical classroom to carry out the experiment individually, is difficult for several reasons, ranging from the fastidious growth requirements of <i>P. calidifontis</i> to the need of specialized equipment for preparing cell extracts. Here, <i>E. coli</i> was used in the overproduction of recombinant <i>P. calidifontis</i> catalase (kat_Pc). The recombinant catalase enzyme was activated with manganese and purified by heat treatment. Experiments discussed here indicate that the use of the recombinant enzyme is a viable alternative for instructors who may or may not have access to <i>P. calidifontis</i> or mechanical means of cell lysis.