How directed evolution reshapes the energy landscape in an enzyme to boost catalysis

Otten, Renee; Pádua, Ricardo A.  P.; Bunzel, H. Adrian; Nguyen, Vy; Pitsawong, Warintra; Patterson, MacKenzie; Sui, Shuo; Perry, Sarah L.; Cohen, Aina E.; Hilvert, Donald; Kern, Dorothee

doi:10.1126/science.abd3623

Title: How directed evolution reshapes the energy landscape in an enzyme to boost catalysis

Journal Article · Fri Dec 18 00:00:00 EST 2020 · Science

DOI:https://doi.org/10.1126/science.abd3623· OSTI ID:1849593

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Howard Hughes Medical Institute and Department of Biochemistry, Brandeis University, Waltham, MA 02454, USA.
Laboratory of Organic Chemistry, ETH Zürich, 8093 Zürich, Switzerland.
Department of Chemical Engineering, Institute of Applied Life Sciences, University of Massachusetts, Amherst, MA 01003, USA.
Stanford Synchrotron Radiation Lightsource, Menlo Park, CA 94025, USA.

Two steps forward—now look back Whether designed computationally or uncovered in activity screening, enzymes repurposed for biocatalysis rarely start at the peak of proficiency. However, directed evolution can in some cases increase catalytic efficiency of a poor enzyme by many orders of magnitude. Ottenet al.used a suite of biochemical techniques to investigate the origins of rate enhancement in a previously evolved model enzyme. Two conformational states are present in the initial, computationally designed enzyme, but only one is active. Shifting the population toward the active state is one factor in increasing catalytic efficiency during evolution. Single mutations do not greatly increase activity, but the synergistic combination of just two out of 17 substitutions can provide most of the rate enhancement seen in the final, evolved enzyme. Science, this issue p.1442

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Research Organization:: Brandeis Univ., Waltham, MA (United States); Univ. of California, Oakland, CA (United States); Stanford Univ., CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC)

DOE Contract Number:: FG02-05ER15699; AC02-05CH11231; AC02-76SF00515

OSTI ID:: 1849593

Journal Information:: Science, Vol. 370, Issue 6523; ISSN 0036-8075

Publisher:: AAAS

Country of Publication:: United States

Language:: English

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