ANCUT2, a Thermo-alkaline Cutinase from Aspergillus nidulans and Its Potential Applications Eva Bermúdez-García 1 & Carolina Peña-Montes 1 & José Augusto Castro-Rodríguez 1 & Augusto González-Canto 2 & Arturo Navarro-Ocaña 1 & Amelia Farrés 1 Received: 11 October 2016 /Accepted: 14 December 2016 # Springer Science+Business Media New York 2017 Abstract Biochemical characterization of purified ANCUT2 cutinase from Aspergillus nidulans is described. The identified amino acid sequence differs from that predicted in Aspergillus genomic databases in amino acids not relevant for catalysis. The enzyme is thermo-alkaline, showing its maximum activity at pH 9 and 60 °C, and it retains more than 60% of its initial activity after incubation for 1 h at 60 °C for pH values between 6 and 10. ANCUT2 is more active towards long-chain esters and it hydrolyzes cutin; however, it also hydro- lyzes short-chain esters. Cutinase is inhibited by metal ions, PMSF, SDS, and EDTA (10 mM). It retains 50% of its activity in most of the solvents tested, although it is more stable in hydrophobic solvents. According to its found biochemical properties, preliminary assays demonstrate its ability to synthesize methyl esters from sesame oil and the most likely application of this enzyme remains in detergent formulations. Keywords Cutinase . Thermo-alkaline . Aspergillus nidulans . Biodiesel . Detergents Appl Biochem Biotechnol DOI 10.1007/s12010-016-2378-z Electronic supplementary material The online version of this article (doi:10.1007/s12010-016-2378-z) contains supplementary material, which is available to authorized users. * Amelia Farrés farres@unam.mx 1 Departamento de Alimentos y Biotecnología, Facultad de Química, Universidad Nacional Autónoma de México (UNAM), Avenida Universidad, 3000 Ciudad Universitaria, 04510 Mexico City, Mexico 2 Departamento de Medicina Experimental, Facultad de Medicina, Hospital General de México, Universidad Nacional Autónoma de México (UNAM), Dr. Balmis, 148, 06726 Mexico City, D.F, Mexico