Reversal of NAD(P)H Cofactor Dependence by Protein Engineering

  • Sabine Bastian
  • Frances H. ArnoldEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 834)


There is increasing interest in utilization of engineered microorganisms for the production of renewable chemicals and next-generation biofuels. However, imbalances between the cofactor consumption of the engineered production pathway and the reducing equivalents provided by the cell have been shown to limit yields. This imbalance can be overcome by adjusting the cofactor dependencies of the pathway enzymes to match the available cofactors in the cell. We show how cofactor preference can be reversed by structure-guided directed evolution of the target enzyme.

Key words

Cofactor switch Enzyme engineering Homology modeling Nicotine amide dinucleotide phosphate Nicotine amide dinucleotide Directed evolution 



The authors would like to thank Dr. Christopher Snow for assistance with homology modeling. This work was sponsored by the U.S. Army Research Laboratory and was accomplished under cooperative Agreement number W911NF-09-2-002.


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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Division of Chemistry and Chemical EngineeringCalifornia Institute of TechnologyPasadenaUSA

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