title: Heterologous expression of an esterase from Thermus thermophilus HB27 in Saccharomyces cerevisiae.
abstract: In this work, a system for high-level secretion by Saccharomyces cerevisiae of the Thermus thermophilus HB27 putative esterase YP_004875.1 was constructed. The recombinant protein was purified and partially characterised. Its lipolytic activity dropped abruptly when the acyl chain length of the substrate increased from 12 to 18 carbon atoms, and variation of the reaction rate as function of substrate concentration followed Michaelis-Menten kinetics. These results suggested that the enzyme was an esterase. The recombinant enzyme was N-glycosylated and both the glycosylated and non-glycosylated forms showed activity. Compared to the native enzyme, thermal stability (half-life of 4.3h at 85 degrees C) was higher, optimum temperature (40 degrees C) was lower and optimum pH (7.5-8.5) was similar. These characteristics support potential biotechnological applications of the recombinant esterase.
authors: Departamento de Bioloxía Celular e Molecular, Facultade de Ciencias, Universidade da Coruña, 15071 A Coruña, Spain.
authors: Olalla
authors: O
authors: López-López
authors: Pablo
authors: P
authors: Fuciños
authors: Lorenzo
authors: L
authors: Pastrana
authors: M Luisa
authors: ML
authors: Rúa
authors: M Esperanza
authors: ME
authors: Cerdán
authors: M Isabel
authors: MI
authors: González-Siso
mesh_terms: D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D004950:Esterases / Q000302:isolation & purification / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008066:Lipolysis; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D012044:Regression Analysis; D012441:Saccharomyces cerevisiae / Q000254:growth & development / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature; D016963:Thermus thermophilus / Q000201:enzymology*
chemical_list: D011994:Recombinant Proteins; D004950:Esterases