Title: Trade-offs between enzyme fitness and solubility illuminated by deep mutational scanning

Abstract

Significance Enzymes find utility as therapeutics and for the production of specialty chemicals. Changing the amino acid sequence of an enzyme can increase solubility, but many such mutations disrupt catalytic activity. To evaluate this trade-off, we developed an experimental system to evaluate the relative solubility for nearly all possible single point mutants for two model enzymes. We find that the tendency for a given solubility-enhancing mutation to disrupt catalytic activity depends, among other factors, on how far the position is from the catalytic active site and whether that mutation has been sampled during evolution. We develop predictive models to identify mutations that enhance solubility without disrupting activity with an accuracy of 90%. These results have biotechnological applications.

Authors:

Klesmith, Justin R.  ^[1];

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Bacik, John-Paul ^[2]; Wrenbeck, Emily E. ^[3]; Michalczyk, Ryszard ^[2]; Whitehead, Timothy A. ^[4]

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+ Show Author Affiliations

1. Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI 48824,
2. Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM 87545,
3. Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing, MI 48824,
4. Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing, MI 48824,, Department of Biosystems and Agricultural Engineering, Michigan State University, East Lansing, MI 48824

Publication Date:

2017-02-14

Sponsoring Org.:

USDOE

OSTI Identifier:

1343782

Grant/Contract Number:  

AC02-76SF00515

Resource Type:

Published Article

Journal Name:

Proceedings of the National Academy of Sciences of the United States of America

Additional Journal Information:

Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Volume: 114 Journal Issue: 9; Journal ID: ISSN 0027-8424

Publisher:

Proceedings of the National Academy of Sciences

Country of Publication:

United States

Language:

English