Grga_2025_Biomol.NMR.Assign_20_3

LCC(ICCG), a bioengineered variant of a cutinase called LCC (Leaf-branch Compost Cutinase), is a high-performance, industrial-grade enzyme capable of efficiently degrading polyethylene terephthalate (PET). This engineered enzyme exhibits significantly enhanced thermal stability and PET hydrolysis activity compared to its predecessor and competing PETases. Here, we report the comprehensive resonance assignment of the polypeptide backbone and the side chain methyl groups of the active LCC(ICCG). Taking advantage of its exceptional thermostability all the experiments were conducted at 60 degreesC on a single, uniformly (15)N-(13)C-labeled sample of this 27 kDa serine-hydrolase enzyme. LCC(ICCG) represents a leap forward in enzymatic PET recycling, combining speed, efficiency, and scalability. The residue-specific information through both backbone and methyl side chain assignment represents a critical step toward detailed structural and dynamic NMR analyses.