caa a22 4500 605504946 CHVBK 20210128100619.0 cr unu---uuuuu 210128e20151201xx s 000 0 eng 10.1007/s00253-015-6890-9 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6890-9 Expression and properties of three novel fungal lipases/sterol esterases predicted in silico: comparison with other enzymes of the Candida rugosa -like family [Elektronische Daten] [María Vaquero, Alicia Prieto, Jorge Barriuso, María Martínez] Lipases from the Candida rugosa-like family are enzymes with great biotechnological interest. In a previous work, several enzymes from this family were identified by in silico mining of fungal genomes. Here, we describe the cloning, expression, and characterization of putative lipases from the genomes of Nectria haematococca, Trichoderma reesei, and Aspergillus niger and compared their catalytic properties with those of OPE, a well-characterized sterol esterase/lipase from Ophiostoma piceae. All of them hydrolyzed p-nitrophenol esters and triglycerides with different efficiency, but their activity against sterol esters was dissimilar, and the enzyme from A. niger was unable of hydrolyzing these substrates while OPE showed the best k cat values, which in general leads to an improved catalytic efficiency. Similarly, OPE was the best catalyst in the synthesis of β-sitostanyl oleate, followed by the commercial CRL from C. rugosa, while the A. niger enzyme was unable to produce this compound. When the enzymes were evaluated for caprolactone oligomerization, the A. niger enzyme gave similar results than CRL, being OPE slightly more efficient. The expression of the putative selected proteins allowed their functional validation, suggesting that the hydrophobicity of the lid region may be an important factor, although the enzymatic efficiency is also influenced by other parameters, as the aggregation state and the size and morphology of the tunnel, where substrate recognition and catalysis takes place. Springer-Verlag Berlin Heidelberg, 2015 Lipases nationallicence Sterol esterases nationallicence Genome mining nationallicence Polycaprolactone nationallicence β-Sitostanol esters nationallicence Vaquero María Department of Environmental Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, E-28040, Madrid, Spain aut Prieto Alicia Department of Environmental Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, E-28040, Madrid, Spain aut Barriuso Jorge Department of Environmental Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, E-28040, Madrid, Spain aut Martínez María Department of Environmental Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, E-28040, Madrid, Spain aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/23(2015-12-01), 10057-10067 0175-7598 99:23<10057 2015 99 253 https://doi.org/10.1007/s00253-015-6890-9 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6890-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Vaquero María Department of Environmental Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, E-28040, Madrid, Spain aut NATIONALLICENCE 700 1- Prieto Alicia Department of Environmental Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, E-28040, Madrid, Spain aut NATIONALLICENCE 700 1- Barriuso Jorge Department of Environmental Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, E-28040, Madrid, Spain aut NATIONALLICENCE 700 1- Martínez María Department of Environmental Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, E-28040, Madrid, Spain aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/23(2015-12-01), 10057-10067 0175-7598 99:23<10057 2015 99 253