Cutinases are hydrolytic enzymes secreted by phytopathogens to degrade cutin, the main polymeric component of plant cuticles. The multifaceted functionality of cutinases has allowed for their exploitation for catalytic reactions beyond their natural purpose. To diversify and expand the cutinase enzyme class, we identified five cutinase homologs from the saprotroph Aspergillus niger. One of these cutinases, AnCUT3, was over-expressed in Pichia pastoris and its biophysicochemical properties characterized. The purified recombinant AnCUT3 possessed an optimum temperature of 25 degreesC, an optimum pH of 5, and was stable at temperatures up to 50 degreesC (1 h incubation, melting point of 45.6 degreesC) and in a wide pH range. Kinetic studies of AnCUT3 using pNP ester substrates showed the highest catalytic efficiency, k(cat)/K(m) of 859 mM(-1) s(-)(1) toward p-nitrophenyl decanoate (C10). Although its calculated molecular mass is 27 kDa, AnCUT3 was expressed as two glycosylated proteins of molecular weights 24 and 50 kDa. Glycan profiling detected the presence of atypical paucimannose N-glycans (>=Man(1-5)GlcNAc) from recombinant AnCUT3, suggesting protein-dependent glycan processing of AnCUT3 in P. pastoris. AnCUT3 was also able to degrade and modify the surface of polycaprolactone and polyethylene terephthalate. Taken together, these features poise AnCUT3 as a potential biocatalyst for industrial applications.
