title: A feruloyl esterase derived from a leachate metagenome library.
abstract: A feruloyl esterase encoding gene (designated fae6), derived from a leachate metagenomic library, was cloned and the nucleotide sequence of the insert DNA determined. Translational analysis revealed that fae6 consists of a 515 amino acid polypeptide, encoding a 55 kDa pre-protein. The Fae6 primary structure contained the G-E-S-A-G sequence, which corresponds well with a typical catalytic serine sequence motif (G-x-S-x-G). The fae6 gene was successfully over-expressed in E. coli and the recombinant protein was purified to 8.4 fold enrichment with 17% recovery. The K(M) data showed Fae6 has a high affinity to methyl sinapate while thermostability data indicated that fae6 was thermolabile with a half life (T(1/2)) < 30 min at 50°C. High affinity for Fae6 against methyl sinapate, methyl ferulate and ethyl ferulate suggest that the enzyme can be useful in hydrolyzing ferulated polysaccharides in a biorefinery process.
authors.affiliation: Enzymes Technologies, CSIR Biosciences, Modderfontein, South Africa. JRashamuse@csir.co.za
authors.forename: Konanani
authors.initials: K
authors.lastname: Rashamuse
authors.forename: Walter
authors.initials: W
authors.lastname: Sanyika
authors.forename: Tina
authors.initials: T
authors.lastname: Ronneburg
authors.forename: Dean
authors.initials: D
authors.lastname: Brady
mesh_terms: D002265:Carboxylic Ester Hydrolases / Q000235:genetics* / Q000302:isolation & purification*; D015723:Gene Library; D054892:Metagenome / Q000235:genetics*; D010802:Phylogeny
chemical_list: D002265:Carboxylic Ester Hydrolases; C070196:feruloyl esterase