TY - JOUR
T1 - Functional enzyme–polymer complexes
AU - Waltmann, Curt
AU - Mills, Carolyn E.
AU - Wang, Jeremy
AU - Qiao, Baofu
AU - Torkelson, John M.
AU - Tullman-Ercek, Danielle
AU - de la Cruz, Monica Olvera
N1 - Funding Information:
ACKNOWLEDGMENTS. We thank the the Department of Energy, Office of Basic Energy Sciences for financial support under Contract DE-FG02-08ER46539. M.O.d.l.C. thanks the Sherman Fairchild Foundation for computational support. C.W., B.Q., J.W., and M.O.d.l.C. thank the Center for Computation and Theory of Soft Materials for support. B.Q. was funded by the Center for Hierarchical Materials Design. J.M.T., M.O.d.l.C., C.E.M. and D.T.-E. were funded by a Cornew Innovation Award through the Chemistry of Life Processes Institute at North-western University. This work made use of the Integrated Molecular Structure Education and Research Center NMR facility at Northwestern University, which has received support from the Soft and Hybrid Nanotechnology Experimental Resource(NSFECCS-2025633)andNorthwesternUniversity.Thisworkalsomade use of the Keck Biophysics Facility (aqueous GPC) at Northwestern University, whichissupportedbytheRobertH.LurieComprehensiveCancerCenter(National Cancer Institute P30 Cancer Center Support Grant CA060553).
Funding Information:
ACKNOWLEDGMENTS. We thank the the Department of Energy, Office of Basic Energy Sciences for financial support under Contract DE-FG02-08ER46539. M.O.d.l.C. thanks the Sherman Fairchild Foundation for computational support. C.W., B.Q., J.W., and M.O.d.l.C. thank the Center for Computation and Theory of Soft Materials for support. B.Q. was funded by the Center for Hierarchical Materials Design. J.M.T., M.O.d.l.C., C.E.M. and D.T.-E. were funded by a Cornew Innovation Award through the Chemistry of Life Processes Institute at Northwestern University. This work made use of the Integrated Molecular Structure Education and Research Center NMR facility at Northwestern University, which has received support from the Soft and Hybrid Nanotechnology Experimental Resource (NSF ECCS-2025633) and Northwestern University. This work also made use of the Keck Biophysics Facility (aqueous GPC) at Northwestern University, which is supported by the Robert H. Lurie Comprehensive Cancer Center (National Cancer Institute P30 Cancer Center Support Grant CA060553).
Publisher Copyright:
© 2022 the Author(s).
PY - 2022/3/29
Y1 - 2022/3/29
N2 - Engineered and native enzymes are poised to solve challenges in medicine, bioremediation, and biotechnology. One important goal is the possibility of upcycling polymers using enzymes. However, enzymes are often inactive in industrial, nonbiological conditions. It is particularly difficult to protect water-soluble enzymes at elevated temperatures by methods that preserve their functionality. Through atomistic and coarse-grained molecular dynamics simulations that capture protein conformational change, we show that an enzyme, PETase (polyethylene terephthalate [PET]), can be stabilized at elevated temperatures by complexation with random copolymers into nanoscale aggregates that do not precipitate into macroscopic phases. We demonstrated the efficiency of the method by simulating complexes of random copolymers and the enzyme PETase, which depolymerizes PET, a highly used polymer. These polymers are more industrially viable than peptides and can target specific domains on an enzyme. We design the mean composition of the random copolymers to control the polymer–enzyme surface contacts and the polymer conformation. When positioned on or near the active site, these polymer contacts can further stabilize the conformation of the active site at elevated temperatures. We explore the experimental implications of this active site stabilization method and show that PETase-random copolymer complexes have enhanced activity on both small molecule substrates and solid PET films. These results provide guidelines for engineering enzyme–polymer complexes with enhanced enzyme functionality in nonbiological environments.
AB - Engineered and native enzymes are poised to solve challenges in medicine, bioremediation, and biotechnology. One important goal is the possibility of upcycling polymers using enzymes. However, enzymes are often inactive in industrial, nonbiological conditions. It is particularly difficult to protect water-soluble enzymes at elevated temperatures by methods that preserve their functionality. Through atomistic and coarse-grained molecular dynamics simulations that capture protein conformational change, we show that an enzyme, PETase (polyethylene terephthalate [PET]), can be stabilized at elevated temperatures by complexation with random copolymers into nanoscale aggregates that do not precipitate into macroscopic phases. We demonstrated the efficiency of the method by simulating complexes of random copolymers and the enzyme PETase, which depolymerizes PET, a highly used polymer. These polymers are more industrially viable than peptides and can target specific domains on an enzyme. We design the mean composition of the random copolymers to control the polymer–enzyme surface contacts and the polymer conformation. When positioned on or near the active site, these polymer contacts can further stabilize the conformation of the active site at elevated temperatures. We explore the experimental implications of this active site stabilization method and show that PETase-random copolymer complexes have enhanced activity on both small molecule substrates and solid PET films. These results provide guidelines for engineering enzyme–polymer complexes with enhanced enzyme functionality in nonbiological environments.
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U2 - 10.1073/pnas.2119509119
DO - 10.1073/pnas.2119509119
M3 - Article
C2 - 35312375
AN - SCOPUS:85126840142
SN - 0027-8424
VL - 119
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 13
M1 - e2119509119
ER -