Carboxyl ester hydrolase from Penicillium expansum : cloning, characterization and overproduction by Penicillium griseoroseum

Abstract

Aims: 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 threedimensional model was obtained. The a/b 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 7Á0 and a temperature of 25°C. CARB exhibited stability in alkaline pH, high activity under mesophilic conditions and stability in organic solvents. Conclusion: 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.

Colletotrichum gloeosporioides lipase: Characterization and use in hydrolysis and esterifications

“…In this case, enzyme preparation showed greater activity than before being stored, suggesting stability of enzyme activity under freezing conditions. These data corroborate results obtained with carboxilesterases stored at low temperature (Corrêa et al, 2013).…”

Production and characterization of a thermostable extracellular esterase from Aspergillus niger

“…Esterases are used in animal feed to enhance digestibility (Bonzom et al, 2018) bio-bleaching of wood-pulp, converting lignocellulosic substances to biofuels and feedstock and processing food to enhance clarification in fruit juices (Correa et al, 2013). These are also used in beverage production (beer and wine), and in synthesis of optically pure compounds, perfumes, and antioxidants (Topakas et al, 2007).…”

Production, purification, and characterization of a novel serine‐esterase from Aspergillus westerdijkiae

“…Furthermore, several esterase genes have been isolated and cloned from metagenomics libraries . However, fewer investigations have been reported on fungal esterases . In this work, we describe the production, the purification, and the characterization of a secreted esterase from Aspergillus westerdijkiae.…”