Arnold lab: coenzyme specificity

The Arnold lab wanted to establish a generalized procedure to change the cofactor specificity of certain enzymes from NADPH to NADH. ^[1]

Active site of the enzyme

The KARI enzyme from S. exigua binds to NADPH. To change the cofactor specificity of the , the researchers took a structure-based directed evolution approach. Try to guess which residues they might have targeted for mutagenesis based on the structure (which was determined after successfully changing the specificity of the enzyme, so they used homology models to figure out which residues are close to the bound cofactor).

ReferencesReferences

↑ Brinkmann-Chen S, Flock T, Cahn JK, Snow CD, Brustad EM, McIntosh JA, Meinhold P, Zhang L, Arnold FH. General approach to reversing ketol-acid reductoisomerase cofactor dependence from NADPH to NADH. Proc Natl Acad Sci U S A. 2013 Jul 2;110(27):10946-51. doi:, 10.1073/pnas.1306073110. Epub 2013 Jun 17. PMID:23776225 doi:10.1073/pnas.1306073110

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Karsten Theis

[1] Brinkmann-Chen S, Flock T, Cahn JK, Snow CD, Brustad EM, McIntosh JA, Meinhold P, Zhang L, Arnold FH. General approach to reversing ketol-acid reductoisomerase cofactor dependence from NADPH to NADH. Proc Natl Acad Sci U S A. 2013 Jul 2;110(27):10946-51. doi:, 10.1073/pnas.1306073110. Epub 2013 Jun 17. PMID:23776225 doi:10.1073/pnas.1306073110

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