Carboxyl ester hydrolase from Penicillium expansum: cloning, characterization and overproduction by Penicillium griseoroseum
Article first published online: 23 APR 2013
Journal of Applied Microbiology © 2013 The Society for Applied Microbiology
Journal of Applied Microbiology
Volume 115, Issue 1, pages 114–124, July 2013
How to Cite
Corrêa, T.L.R., Zubieta, M.P., Teixeira, J.A., de Queiroz, M.V. and de Araújo, E.F. (2013), Carboxyl ester hydrolase from Penicillium expansum: cloning, characterization and overproduction by Penicillium griseoroseum. Journal of Applied Microbiology, 115: 114–124. doi: 10.1111/jam.12215
- Issue published online: 12 JUN 2013
- Article first published online: 23 APR 2013
- Accepted manuscript online: 12 APR 2013 09:32AM EST
- Manuscript Accepted: 1 APR 2013
- Manuscript Revised: 27 MAR 2013
- Manuscript Received: 7 MAR 2013
- Brazilian Foundation for Research Support of Minas Gerais (FAPEMIG)
- National Council for Scientific and Technological Development (CNPq)
- enzyme characterization;
- heterologous expression;
- Penicillium expansum ;
- Penicillium griseoroseum
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.
Methods and 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 Ser213-Glu341-His456 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.
The CARB protein is potentially useful in bioremediation, food and chemical/pharmaceutical industries.
Significance and Impact of the Study
This study is the first to report the development of a recombinant strain superproducing a Penicillium sp. carboxylesterase.