Poster
Presentation 1B-26
Thermostability
of a Hybrid Cytochrome P450 Enzyme Generated from CYP101 and CYP119
Abhijeet P. Borole* and Choo Y. Hamilton
Life Sciences Division
Oak Ridge National Laboratory
Bldg. 4505, MS 6226
Phone: (865)576-7421
Fax:
(865)574-6442
E-mail: borolea@ornl.gov
Cytochrome
P450 is one of the most versatile enzymes for oxygen activation of hydrophobic
molecules such as polyaromatic hydrocarbons and alkanes. Industrial application of these enzymes has
been limited by their low stability and requirement for a cofactor. In an attempt to increase thermostability of
these enzymes, we are studying shuffling and chimeragenesis with related
thermostable enzymes. CYP101 is our model hydrocarbon oxidizing enzyme and
CYP119 is the model thermostable enzyme. Chimeras generated from these two proteins
show retention of the temperature stability but alteration in substrate
specificity. Preliminary results from a
combined rational design followed by random shuffling strategy to produce a
functional thermostable protein will be discussed.