TY - JOUR
T1 - Designed CYP450 Scaffold That Possesses a Local Electric Field Biodegrades Polyethylene Terephthalate
AU - Siddiqui, Shakir Ali
AU - Shaik, Sason
AU - Dubey, Kshatresh Dutta
N1 - Publisher Copyright:
© 2024 American Chemical Society.
PY - 2024/10/18
Y1 - 2024/10/18
N2 - In enzymes, protein residues themselves carry charges and produce a preorganized local electric field (LEF). Such LEFs can be modified by strategically mutating the charged and polar residues to create a designed LEF (D-LEF) to project on the reaction axis to modulate the reactivity of the enzyme. We investigated the enzymatic degradation of polyethylene terephthalate (PET) using in silico engineering of CYP450 enzymes, which possess D-LEFs. We show that PET degradation can, in principle, be catalyzed using a CYP450 scaffold through oxidative cleavage of the ester bond. The PET degradation occurs in two crucial steps; the first step is the usual hydroxylation reaction initiated by compound I (Cpd I), while the second step is the dealkylation of the gem-hydroxy moiety, which is driven by the preorganized LEF of the CYP450 enzyme. Using molecular-level analyses for three different enzymes, we found that each of the three can efficiently catalyze the HAT reaction. However, only CYP450GcoA performs an efficient dealkylation reaction since it possesses the only scaffold among the three enzymes that has preorganized LEF properly oriented for dealkylation. We show that a strategic mutation based on the designed LEF along the reaction axis and the binding site architecture can evolve the enzyme for the PET degradation reaction. Our study further provides a key lesson that intuiting the LEF of the enzyme in the direction of the reaction axis could be crucial in selecting the most suitable scaffold for the desired reaction.
AB - In enzymes, protein residues themselves carry charges and produce a preorganized local electric field (LEF). Such LEFs can be modified by strategically mutating the charged and polar residues to create a designed LEF (D-LEF) to project on the reaction axis to modulate the reactivity of the enzyme. We investigated the enzymatic degradation of polyethylene terephthalate (PET) using in silico engineering of CYP450 enzymes, which possess D-LEFs. We show that PET degradation can, in principle, be catalyzed using a CYP450 scaffold through oxidative cleavage of the ester bond. The PET degradation occurs in two crucial steps; the first step is the usual hydroxylation reaction initiated by compound I (Cpd I), while the second step is the dealkylation of the gem-hydroxy moiety, which is driven by the preorganized LEF of the CYP450 enzyme. Using molecular-level analyses for three different enzymes, we found that each of the three can efficiently catalyze the HAT reaction. However, only CYP450GcoA performs an efficient dealkylation reaction since it possesses the only scaffold among the three enzymes that has preorganized LEF properly oriented for dealkylation. We show that a strategic mutation based on the designed LEF along the reaction axis and the binding site architecture can evolve the enzyme for the PET degradation reaction. Our study further provides a key lesson that intuiting the LEF of the enzyme in the direction of the reaction axis could be crucial in selecting the most suitable scaffold for the desired reaction.
KW - enzyme engineering
KW - local electric field (LEF)
KW - MD simulations
KW - PET degradation
KW - QM/MM calculations
UR - http://www.scopus.com/inward/record.url?scp=85205904695&partnerID=8YFLogxK
U2 - 10.1021/acscatal.4c05363
DO - 10.1021/acscatal.4c05363
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AN - SCOPUS:85205904695
SN - 2155-5435
VL - 14
SP - 15108
EP - 15122
JO - ACS Catalysis
JF - ACS Catalysis
IS - 20
ER -