Massively used plastics have caused worldwide environmental concerns. Polyesters like polylactic acid (PLA) are one of the mostly used plastics due to its excellent physical and chemical properties and low-cost advantages. It is critical to develop the elimination and recycle techniques for polyesters. Experimental studies have shown that a hydrolase RPA1511 isolated from Rhodopseudomonas palustris can efficiently depolymerize polylactic acid (PLA) into oligomers and monomers. It was also active against emulsified aliphatic polymers as well as multipurpose soluble ester monomers (α-naphthyl ester and p-nitrophenyl ester). In the present study, molecular dynamics simulations and molecular mechanics Poisson-Boltzmann surface area method were applied to screen all amino acids from hydrolase RPA1511 and identify the most important amino acids during substrate binding. Seven substrates were considered: PLA (dimer and tetramer), polycaprolactone, butylene succinate, 1-naphthyl acetate, 2-naphthyl formate, p-nitrophenyl acetate. The results highlighted the importance of amino acids like Tyr139, Tyr213, Arg259, Thr46. Subsequent quantum mechanics/molecular mechanics calculations were also performed to determine the detailed degradation mechanism of hydrolase RPA1511 toward PLA and explore the role of the active site residues during catalysis. The results demonstrated that degradation involves two elementary steps: enzyme acylation and PLA hydrolysis. The corresponding Boltzmann average barriers are 20.40 kcal/mol and 14.45 kcal/mol. The electrostatic influence analysis of 15 amino acids on the rate-determining step indicated that amino acids His114, Trp219 and Ala273 facilitate the reaction while the Arg244 suppresses the reaction which may serve as future mutation studies to enhance the enzymatic efficiency.
Keywords: Enzymatic transformation; MM-PBSA; Polyester; Quantum mechanics/molecular mechanics.
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