Purification, characterization and mass spectrometric sequencing of a thermophilic glucuronoyl esterase from Sporotrichum thermophile

Vafiadi, C; Topakas, E; Biely, P; Christakopoulos, P

dc.contributor.author	Vafiadi, C	en
dc.contributor.author	Topakas, E	en
dc.contributor.author	Biely, P	en
dc.contributor.author	Christakopoulos, P	en
dc.date.accessioned	2014-03-01T01:31:44Z
dc.date.available	2014-03-01T01:31:44Z
dc.date.issued	2009	en
dc.identifier.issn	0378-1097	en
dc.identifier.uri	https://dspace.lib.ntua.gr/xmlui/handle/123456789/19911
dc.subject	Glucuronoyl esterase	en
dc.subject	Lignin-carbohydrate complex	en
dc.subject	Mass spectrometric sequencing	en
dc.subject	Purification	en
dc.subject	Sporotrichum thermophile	en
dc.subject.classification	Microbiology	en
dc.subject.other	esterase	en
dc.subject.other	glucuronoyl esterase 1	en
dc.subject.other	pyranoside	en
dc.subject.other	unclassified drug	en
dc.subject.other	article	en
dc.subject.other	catalysis	en
dc.subject.other	enzyme activity	en
dc.subject.other	enzyme analysis	en
dc.subject.other	enzyme purification	en
dc.subject.other	enzyme stability	en
dc.subject.other	enzyme synthesis	en
dc.subject.other	fungus	en
dc.subject.other	gene sequence	en
dc.subject.other	mass spectrometry	en
dc.subject.other	mesophile	en
dc.subject.other	molecular weight	en
dc.subject.other	nonhuman	en
dc.subject.other	nucleotide sequence	en
dc.subject.other	pH	en
dc.subject.other	priority journal	en
dc.subject.other	sequence homology	en
dc.subject.other	sporotrichum thermophile	en
dc.subject.other	temperature	en
dc.subject.other	Amino Acid Sequence	en
dc.subject.other	Enzyme Stability	en
dc.subject.other	Fungal Proteins	en
dc.subject.other	Glucuronates	en
dc.subject.other	Glucuronidase	en
dc.subject.other	Hydrogen-Ion Concentration	en
dc.subject.other	Isoelectric Point	en
dc.subject.other	Lignin	en
dc.subject.other	Mass Spectrometry	en
dc.subject.other	Molecular Sequence Data	en
dc.subject.other	Molecular Weight	en
dc.subject.other	Sequence Homology, Amino Acid	en
dc.subject.other	Sporothrix	en
dc.subject.other	Substrate Specificity	en
dc.subject.other	Temperature	en
dc.subject.other	Xylans	en
dc.subject.other	Chaetomium globosum CBS 148.51	en
dc.subject.other	Corynascus heterothallicus	en
dc.subject.other	Fungi	en
dc.title	Purification, characterization and mass spectrometric sequencing of a thermophilic glucuronoyl esterase from Sporotrichum thermophile	en
heal.type	journalArticle	en
heal.identifier.primary	10.1111/j.1574-6968.2009.01631.x	en
heal.identifier.secondary	http://dx.doi.org/10.1111/j.1574-6968.2009.01631.x	en
heal.language	English	en
heal.publicationDate	2009	en
heal.abstract	The cellulolytic system of the thermophilic fungus Sporotrichum thermophile contains a recently discovered esterase that may hydrolyze the ester linkage between the 4-O-methyl-d-glucuronic acid of glucuronoxylan and lignin alcohols. The glucuronoyl esterase named StGE1 was purified to homogeneity with a molecular mass of Mr 58 kDa and pI 6.7. The enzyme activity was optimal at pH 6.0 and 60 °C. The esterase displayed a narrow pH range stability at pH 8.0 and retained 50% of its activity after 430 and 286 min at 50 and 55 °C, respectively. The enzyme was active on substrates containing glucuronic acid methyl ester, showing a lower catalytic efficiency on 4-nitrophenyl 2-O-(methyl-4-O-methyl-α-d-glucopyranosyluronate)-β-d- xylopyranoside than its mesophilic counterparts reported in the literature, which is typical of thermophilic enzymes. StGE1 was proved to be a modular enzyme containing a noncatalytic carbohydrate-binding module. LC-MS/MS analysis provided peptide mass and sequence information that facilitated the identification and classification of StGE1 as a family 15 glucuronoyl esterase that showed the highest homology with the hypothetical glucuronoyl esterase CHGG-10774 of Chaetomium globosum CBS 148.51. This work represents the first example of the purification and identification of a thermophilic glucuronoyl esterase from S. thermophile. © 2009 Federation of European Microbiological Societies.	en
heal.publisher	WILEY-BLACKWELL PUBLISHING, INC	en
heal.journalName	FEMS Microbiology Letters	en
dc.identifier.doi	10.1111/j.1574-6968.2009.01631.x	en
dc.identifier.isi	ISI:000266979400005	en
dc.identifier.volume	296	en
dc.identifier.issue	2	en
dc.identifier.spage	178	en
dc.identifier.epage	184	en