title: Carboxyl ester hydrolase from Penicillium expansum: cloning, characterization and overproduction by Penicillium griseoroseum.
abstract: OBJECTIVE
  In this study, a gene that encodes a carboxylesterase (carb) in Penicillium expansum GF was cloned, sequenced and overexpressed by Penicillium griseoroseum PG63, and the enzyme was characterized.
  
  
  RESULTS
  The recombinant strain, P. griseoroseum T55, obtained upon transformation using the plasmid pAN-52-1-carb, showed integration of the carb gene into at least two heterologous sites of the genome by Southern blotting. Furthermore, the recombinant strain T55 exhibited almost a fourfold increase in carboxylesterase activity compared with PG63 strain when both were cultured without inducers. Based on the secondary structure and multiple sequence alignments with carboxylesterases, cholinesterase and lipase, a three-dimensional model was obtained. The α/β barrel topology, that is typical of esterases and lipases, was indicated for the CARB protein with Ser(213)-Glu(341)-His(456) as the putative catalytic triad. CARB preferentially hydrolysed acyl chains with eight carbon atoms, and its activity was optimal at a pH of 7·0 and a temperature of 25°C. CARB exhibited stability in alkaline pH, high activity under mesophilic conditions and stability in organic solvents.
  
  
  CONCLUSIONS
  The CARB protein is potentially useful in bioremediation, food and chemical/pharmaceutical industries.
  
  
  CONCLUSIONS
  This study is the first to report the development of a recombinant strain superproducing a Penicillium sp. carboxylesterase.
authors.affiliation: Department of Microbiology/BIOAGRO, Federal University of Viçosa (Universidade Federal de Viçosa), Viçosa, MG, Brazil.
authors.forename: T L R
authors.initials: TL
authors.lastname: Corrêa
authors.forename: M P
authors.initials: MP
authors.lastname: Zubieta
authors.forename: J A
authors.initials: JA
authors.lastname: Teixeira
authors.forename: M V
authors.initials: MV
authors.lastname: de Queiroz
authors.forename: E F
authors.initials: EF
authors.lastname: de Araújo
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D043182:Carboxylesterase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D008969:Molecular Sequence Data; D010407:Penicillium / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment
chemical_list: D043182:Carboxylesterase