A spectrophotometric assay for feruloyl esterases

Modification of wheat straw lignin by solid state fermentation with white-rot fungi

“…Hydroxycinnamic acids, particularly ferulic acid and p-coumaric acid ( Figure 1.4), and to some extent, caffeic and sinapic acids occur widely in the cell walls of wheat straw (Benoit et al, 2006) and their probable role is to decrease cell wall biodegradability and regulate the growth by crosslinking of cell wall polymers (Mastihuba et al, 2002). Furthermore, phenolic cross-links using ferulic or p-coumaric acids contribute to the Modification of wheat straw lignin by solid state fermentation with white-rot fungi 7 structural integrity of the cell wall by mediating direct covalent interactions between hemicellulose chains and lignin (Panagiotou et al, 2007;Buranov et al, 2008).…”

A practical high-throughput screening system for feruloyl esterases: Substrate design and evaluation

“…To evaluate the feasibility of the four mimic substrates for the high-throughput screening system of FAEs in a 96-well microplate format, the reaction mixture was prepared by mixing 160 L of 100 mM sodium phosphate buffer (pH 6.4) containing 2.5% Triton X-100 with 10 L of 20 mM substrate in DMSO [10], followed by the addition of 20 L supernatant and sufficient amount of ␣-l-arabinofuranosidase (␣-AF) (Megazyme, Wicklow, Ireland) only in case of CNPFA and UFA. The reactions were performed in triplicate at 40 • C for 10 min with shaking at 100 rpm.…”

“…The assay of FAEs' activity commonly depends on HPLC, using enzymatic hydrolysis of a variety of natural and synthetic compounds, such as hydroxycinnamic esterified polysaccharide, methyl feulate and their analogues [6][7][8][9]. Recently, several chlorogenic substrates of FAEs have been reported for quantitative assays, such as 4-nitrophenyl ferulate [10,11] (Fig. 1), 2-O-and 5-Oferuloylated 4-nitrophenyl-l-arabinofuranosides [12], feruloylated p-nitrophenyl ␣-l-arabinofuranosides and ␤-d-xylopyranosides [13], and 2-chloro-4-nitrophenyl 5-O-hydroxycinnamoyl-␣-larabinofuranoside ( Fig.…”

Identification and Characterization of Feruloyl Esterases Produced by Probiotic Bacteria

“…Using absorbance to obtain enzyme kinetic parameters from phenolic ester hydrolysis can be difficult. An alternative method such as high performance liquid chromatography could be used to estimate the enzymatic activity by monitoring the release of products during phenolic ester hydrolysis (Mastihuba et al, 2002).…”