Heterologous production of the Piromyces equi cinnamoyl esterase in Trichoderma reesei for biotechnological applications.
OBJECTIVE
The objective of the study was to produce and characterize the cinnamoyl esterase EstA from the anaerobic fungus Piromyces equi for potential industrial applications.


RESULTS
The catalytic domain EstA was produced in Trichoderma reesei. Because the two fungi displayed different genome features, including different codon usage and GC content, a synthetic gene was designed and expressed, leading to the production of the corresponding protein at around 33 mg per litre in the T. reesei culture medium. After the recombinant protein was purified, biochemical characterization showed that EstA presents peak activity at pH 6.5 and at 50-60 degrees C. Furthermore, EstA remained stable at pH 6-8 and below 50 degrees C. EstA was compared to cinnamoyl esterases FaeA and FaeB from Aspergillus niger in terms of ferulic acid (FA) release from wheat bran (WB), maize bran (MB) and sugar beet pulp (SBP).


CONCLUSIONS
The synthetic gene was successfully cloned and overexpressed in T. reesei. EstA from P. equi was demonstrated to efficiently release FA from various natural substrates.


CONCLUSIONS
Recombinant EstA produced in an industrial enzyme producer, T. reesei, was biochemically characterized, and its capacity to release an aromatic compound (FA) for biotechnological applications was demonstrated.
IFP, Institut Français du Pétrole, Rueil-Malmaison, France. laetitia.poidevin@esil.univmed.fr
L
L
Poidevin
A
A
Levasseur
G
G
Paës
D
D
Navarro
S
S
Heiss-Blanquet
M
M
Asther
E
E
Record
D001234:Aspergillus niger / Q000201:enzymology; D002265:Carboxylic Ester Hydrolases / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D003373:Coumaric Acids / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007218:Industrial Microbiology; D020089:Piromyces / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature; D014242:Trichoderma / Q000235:genetics / Q000378:metabolism*
D003373:Coumaric Acids; D005656:Fungal Proteins; D011994:Recombinant Proteins; C004999:ferulic acid; D002265:Carboxylic Ester Hydrolases; C070196:feruloyl esterase