Purification and characterization of a type B feruloyl esterase (StFAE-A) from the thermophilic fungus Sporotrichum thermophile
| dc.contributor.author | Topakas, E. | en |
| dc.contributor.author | Stamatis, H. | en |
| dc.contributor.author | Biely, P. | en |
| dc.contributor.author | Christakopoulos, P. | en |
| dc.date.accessioned | 2015-11-24T16:33:27Z | |
| dc.date.available | 2015-11-24T16:33:27Z | |
| dc.identifier.issn | 0175-7598 | - |
| dc.identifier.uri | https://olympias.lib.uoi.gr/jspui/handle/123456789/7663 | |
| dc.rights | Default Licence | - |
| dc.subject | 1-Butanol/metabolism | en |
| dc.subject | Caffeic Acids/metabolism | en |
| dc.subject | Carboxylic Ester Hydrolases/*isolation & purification/*metabolism | en |
| dc.subject | Chromatography, Gel | en |
| dc.subject | Chromatography, Ion Exchange | en |
| dc.subject | Coumaric Acids/metabolism | en |
| dc.subject | Dietary Fiber/metabolism | en |
| dc.subject | Dimerization | en |
| dc.subject | Enzyme Stability | en |
| dc.subject | He | en |
| dc.title | Purification and characterization of a type B feruloyl esterase (StFAE-A) from the thermophilic fungus Sporotrichum thermophile | en |
| heal.abstract | A feruloyl esterase (StFAE-A) produced by Sporotrichum thermophile was purified to homogeneity. The purified homogeneous preparation of native StFAE-A exhibited a molecular mass of 57.0+/-1.5 kDa, with a mass of 33+/-1 kDa on SDS-PAGE. The pI of the enzyme was estimated by cation-exchange chromatofocusing to be at pH 3.1. The enzyme activity was optimal at pH 6.0 and 55-60 degrees C. The purified esterase was stable at the pH range 5.0-7.0. The enzyme retained 70% of activity after 7 h at 50 degrees C and lost 50% of its activity after 45 min at 55 degrees C and after 12 min at 60 degrees C. Determination of k(cat)/ K(m) revealed that the enzyme hydrolyzed methyl p-coumarate 2.5- and 12-fold more efficiently than methyl caffeate and methyl ferulate, respectively. No activity on methyl sinapinate was detected. The enzyme was active on substrates containing ferulic acid ester linked to the C-5 and C-2 linkages of arabinofuranose and it hydrolyzed 4-nitrophenyl 5- O- trans-feruloyl-alpha- l-arabinofuranoside (NPh-5-Fe-Ara f) 2-fold more efficiently than NPh-2-Fe-Ara f. Ferulic acid (FA) was efficiently released from destarched wheat bran when the esterase was incubated together with xylanase from S. thermophile (a maximum of 34% total ferulic acid released after 1 h incubation). StFAE-A by itself could release FA, but at a level almost 47-fold lower than that obtained in the presence of xylanase. The potential of StFAE-A for the synthesis of various phenolic acid esters was tested using a ternary water-organic mixture consisting of n-hexane, 1-butanol and water as a reaction system. | en |
| heal.access | campus | - |
| heal.fullTextAvailability | TRUE | - |
| heal.identifier.primary | 10.1007/s00253-003-1481-6 | - |
| heal.identifier.secondary | http://www.ncbi.nlm.nih.gov/pubmed/14615854 | - |
| heal.identifier.secondary | http://www.springerlink.com/content/n5a3qa4c0ceafeqb/fulltext.pdf | - |
| heal.journalName | Appl Microbiol Biotechnol | en |
| heal.journalType | peer reviewed | - |
| heal.language | en | - |
| heal.publicationDate | 2004 | - |
| heal.recordProvider | Πανεπιστήμιο Ιωαννίνων. Σχολή Επιστημών και Τεχνολογιών. Τμήμα Βιολογικών Εφαρμογών και Τεχνολογιών | el |
| heal.type | journalArticle | - |
| heal.type.el | Άρθρο Περιοδικού | el |
| heal.type.en | Journal article | en |
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