Purification and characterization of a Fusarium oxysporum feruloyl esterase (FoFAE-I) catalysing transesterification of phenolic acid esters

 
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Πανεπιστήμιο Ιωαννίνων
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Ιδρυματικό Αποθετήριο Ολυμπιάς
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Purification and characterization of a Fusarium oxysporum feruloyl esterase (FoFAE-I) catalysing transesterification of phenolic acid esters (EN)

Topakas, E. (EN)

Πανεπιστήμιο Ιωαννίνων. Σχολή Επιστημών και Τεχνολογιών. Τμήμα Βιολογικών Εφαρμογών και Τεχνολογιών (EL)
Topakas, E. (EN)

An extracellular feruloyl esterase (FoFAE-I) from the culture filtrates of Fusarium oxysporum F3 was purified to homogeneity by ion-exchange, hydrophobic interaction and gel filtration chromatographies. The protein corresponded to molecular mass and pI values of 31 kDa and 9.5, respectively. The enzyme was optimally active at pH 7.0 and 55 degreesC. The purified esterase was fully stable at pH 7.0-9.0 and temperature up to 30 degreesC. Determination of k(cat)/K-m revealed that the enzyme hydrolysed methyl p-coumarate (MpCA) 4.5, 9, and 239 times more efficiently than methyl caffeate (MCA), methyl ferulate (MFA) and methyl sinapinate (MSA), respectively. The enzyme was active on substrates containing ferulic acid ester linked to the C-5 and C-2 linkages of arabinofuranose but showed preference for the ester at position 2. 4-Nitrophenyl-2-O-trans-feruloyl-alpha-L-arabinofuranoside (NPh-5-Fe-Araf) was hydrolysed 100 times more efficiently than 4-nitrophenyl-5-O-trans-feruloyl-alpha-L-arabinofuranoside (NPh-2-Fe-Araf). Ferulic acid (FA) was efficiently released from destarched wheat bran (DSWB) when the esterase was incubated together with xylanase from Sporotrichum thermophile (a maximum of 92% total ferulic acid released after 4 h incubation). FoFAE-I by itself could release FA but at a level almost five-fold lower than that obtained in the presence of xylanase. The potential of FAE-I for the synthesis of various phenolic acid esters was tested using as a reaction system a surfactantless microemulsions formed in ternary mixture consisting of n-hexane, 1-butanol and water. (C) 2003 Elsevier Inc. All rights reserved. (EN)

fusarium oxysporum (EN)

Πανεπιστήμιο Ιωαννίνων (EL)
University of Ioannina (EN)

Enzyme and Microbial Technology (EN)

Αγγλική γλώσσα

2003

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