Title : Purification, characterization and mass spectrometric sequencing of a thermophilic glucuronoyl esterase from Sporotrichum thermophile. - Vafiadi_2009_FEMS.Microbiol.Lett_296_178 |
Author(s) : Vafiadi C , Topakas E , Biely P , Christakopoulos P |
Ref : FEMS Microbiology Letters , 296 :178 , 2009 |
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 M(r) 58 kDa and pI 6.7. The enzyme activity was optimal at pH 6.0 and 60 degrees 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 degrees 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-alpha-d-glucopyranosyluronate)-beta-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. |
PubMedSearch : Vafiadi_2009_FEMS.Microbiol.Lett_296_178 |
PubMedID: 19459957 |
Gene_locus related to this paper: chagb-Q2GMN0 , thiha-cip2 |
Substrate | NPh-Xyl-MeGlcA |
Gene_locus | chagb-Q2GMN0 thiha-cip2 |
Family | Glucuronoyl_esterase |
Structure | 4G4G 4G4I 4G4J |
Vafiadi C, Topakas E, Biely P, Christakopoulos P (2009)
Purification, characterization and mass spectrometric sequencing of a thermophilic glucuronoyl esterase from Sporotrichum thermophile.
FEMS Microbiology Letters
296 :178
Vafiadi C, Topakas E, Biely P, Christakopoulos P (2009)
FEMS Microbiology Letters
296 :178