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A General Tool for Engineering the NAD/NADP Cofactor Preference of Oxidoreductases by Cahn, Jackson Kenai Blender; Werlang, Caroline A.; Baumschlager, Armin; Brinkmann-Chen, Sabine; Mayo, Stephen Leon; Arnold, Frances H. is a nonfiction available to read on EtoBox.

What is A General Tool for Engineering the NAD/NADP Cofactor Preference of Oxidoreductases about?

The ability to control enzymatic nicotinamide cofactor utilization is critical for engineering efficient metabolic pathways. However, the complex interactions that determine cofactor-binding preference render this engineering particularly challenging. Physics-based models have been insufficiently accurate and blind directed evolution methods too inefficient to be widely adopted. Building on a comprehensive survey of previous studies and our own prior engineering successes, we present a structure-guided, semirational strategy for reversing enzymatic nicotinamide cofactor specificity. This heuristic-based approach leverages the diversity and sensitivity of catalytically productive cofactor binding geometries to limit the problem to an experimentally tractable scale. We demonstrate the efficacy of this strategy by inverting the cofactor specificity of four structurally diverse NADP-dependent enzymes: glyoxylate reductase, cinnamyl alcohol dehydrogenase, xylose reductase, and iron-containing alcohol dehydrogenase. The analytical components of this approach have been fully automated and are available in the form of an easy-to-use web tool: Cofactor Specificity Reversal-Structural Analys

Who reads A General Tool for Engineering the NAD/NADP Cofactor Preference of Oxidoreductases?

Author

Cahn, Jackson Kenai Blender; Werlang, Caroline A.; Baumschlager, Armin; Brinkmann-Chen, Sabine; Mayo, Stephen Leon; Arnold, Frances H.

Publisher

American Chemical Society (ACS)

Published

2016

Language

EN

Category

nonfiction

Subjects

Science, Engineering, Stem