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Versatile capacity of shuffled cytochrome P450s for dye production

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Abstract

DNA family shuffling is a relatively new method of directed evolution used to create novel enzymes in order to improve their existing properties or to develop new features. This method of evolution in vitro has one basic requirement: a high similarity of initial parental sequences. Cytochrome P450 enzymes are relatively well conserved in their amino acid sequences. Members of the same family can have more than 40% of sequence identity at the protein level and are therefore good candidates for DNA family shuffling. These xenobiotic-metabolising enzymes have an ability to metabolise a wide range of chemicals and produce a variety of products including blue pigments such as indigo. By applying the specifically designed DNA family shuffling approach, catalytic properties of cytochrome P450 enzymes were further extended in the chimeric progeny to include a new range of blue colour formations. This mini-review evokes the possibility of exploiting directed evolution of cytochrome P450s and the novel enzymes created by DNA family shuffling for the production of new dyes.

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Acknowledgments

The author would like to thank Drs. Elizabeth Gillam, James De Voss, Pavel Soucek, Robert Kinobe, Deanne Mitchell, Shona Osborne and Iris Depaz for their help with different aspects of this research. International Postgraduate Research Scholarship and University of Queensland Graduate School Award (N. R.) also supported this project.

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Correspondence to Nedeljka N. Rosic.

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Rosic, N.N. Versatile capacity of shuffled cytochrome P450s for dye production. Appl Microbiol Biotechnol 82, 203–210 (2009). https://doi.org/10.1007/s00253-008-1812-8

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Keywords

  • Directed evolution
  • DNA family shuffling
  • CYP
  • Indole metabolism
  • Molecular breeding
  • Indigo