Characterization of a feruloyl esterase from aspergillus. Purification and biochemical characterization of ferulic. This is consistent with the results above, which show that ferulic acid either stimulated or derepressed feruloyl esterase production by a. Release of ferulic acid from oat hulls by aspergillus ferulic. An inducible ferulic acid esterase faeiii has been isolated, purified and partially characterized from aspergillus niger after growth on oat spelt xylan. Pdf an inducible ferulic acid esterase faeill has been isolated, purified and partially characterized from aspergillus niger after growth on oat. Production of feruloyl esterase from aspergillus niger by solidstate.
Iii accounted for most of the feruloyl esterase activity. Pdf release of ferulic acid from wheat bran by a ferulic. Expanding the feruloyl esterase gene family of aspergillus niger by. Two forms of ferulic acid esterase from aspergillus niger have been isolated from a commercial source of pectinase. The major focus of the present work was to study ferulic acid esterase from a strain of aspergillus niger cfr 1105 with respect to a optimization of growth conditions of aspergillus niger cfr 1105 for the enrichment of ferulic acid esterase, b its purification and partial characterization. It also shows that other esterlinked groups, such as acetyl or methoxy groups, are not essential for feruloyl esterase production. Pdf the crystal structure of feruloyl esterase a from. Pentylferulate synthesis was achieved at high yields 5060% with aspergillus niger feruloyl esterase using a waterinoil microemulsion system. Feruloyl esterases are enzymes that release aromatic residues, such as ferulic acid, from plant cell wall polymers 5. Characterization of a feruloyl esterase from aspergillus terreus facilitates the division of fungal enzymes from carbohydrate esterase family 1 of the carbohydrateactive enzymes cazy database miia r. In this paper, we examine both the ability of an a. Aspergillus niger first transformed ferulic acid into vanillic acid, and vanillic acid was then metabolized into vanillin by pycnoporus cinnabarinus. Apr 15, 2012 ferulic acid fa is a component of plant cell walls that has applications in food, cosmetic, and health products, but its applications are limited by its high insolubility.
The feruloyl esterase a faeaencoding gene faea from aspergillus niger has been cloned, and induction of this gene has been studied at the protein level 5. Highlevel production of recombinant aspergillus niger cinnamoyl esterase faea in the methylotrophic yeast. Influence of different xylanases on the activity of ferulic. Pdf an aspergillus niger esterase ferulic acid esterase. Pdf the crystallographic structure of feruloyl esterase from aspergillus niger has been determined to a resolution of 1. Probing the determinants of substrate specificity of a. An enzyme system is described that is useful for preparing food and feed. Purification and biochemical characterization of ferulic acid. The purification procedure utilized ammonium sulphate precipitation, hydrophobic interaction and. Chemical and thermal stability of ferulic acid feruloyl. A feruloyl esterase fae from aspergillus niger n402, faec was heterologously.
Expression of the feruloyl esterase encoding gene faea from aspergillus niger depends on dxylose expression is mediated by xlnr, the xylanolytic transcriptional activator and on a second system that. Ferulic acid was efficiently released from a wheat bran preparation by a ferulic acid esterase from aspergillus niger faeiii when incubated together with a trichoderma viride xylanase a. The enzyme having esterification activity was isolated and was found to have a specific activity of 9. Pdf structure of a feruloyl esterase from aspergillus niger. Expanding the feruloyl esterase gene family of aspergillus. The purified enzyme appeared almost pure by sdspage, with an apparent m r of 36000. Visserthe fae a genes from aspergillus niger and aspergillus tubingensis encode ferulic acid esterases involved in degradation of complex cell wall polysaccharides. A feruloyl esterase from aspergillus niger anfaea in complex with a feruloylated trisaccharide substrate. Production of feruloyl esterase from aspergillus niger by. Purification and characterization of a ferulic acid esterase faeiii from aspergillus niger. Influence of ferulic acid on the production of feruloyl esterases by. Release of ferulic acid from wheat bran by a ferulic acid.
Expression of a fungal ferulic acid esterase in alfalfa. We isolated a ferulic acid esterase faea from a commer cial pectinase preparation which is similar to faeiii 14. Influence of ferulic acid on feruloyl esterase and xylanase production. In this study, we found that a commercial preparation of pectinase pl amano prepared from a. This enzyme belongs to the family of hydrolases, specifically those acting on carboxylic ester bonds. A high level of faea activity in the culture filtrate was detected when an overproducing transformant was. Us6143543a enzyme system comprising ferulic acid esterase. The faea genes from aspergillus niger and aspergillus. Solid state fermentation ssf culture conditions were investigated for the production of feruloyl esterase by aspergillus niger i1472 and compared with submerged culture conditions. Specificity for the phenolic moiety and binding to microcrystalline cellulose. Targeting expression of a fungal ferulic acid esterase to.
Structure of a feruloyl esterase from aspergillus niger. Here we describe the crystal structure of inactive s3a mutant of type. Isolation and characterization of a ferulic acid esterase fae1a from. Synthesis of highly watersoluble feruloyl diglycerols by. Influence of ferulic acid on the production of feruloyl esterases by aspergillus niger. Pdf release of ferulic acid from wheat bran by ferulic. The vast majority of anfaea transformed plant lines showed significant decreases in the cell wall content of. Synthesis of pentylferulate by a feruloyl esterase from. Phenolics also limit the degradation of plant cell walls by saprophytic microbes and by anaerobic microorganisms in the rumen. Addition of free ferulic acid to osx at the start of the culture induced fae. Pdf production of a chimeric enzyme tool associating the. Only the ferulic acid moiety of the substrate is visible in the electron density map, showing interactions through its oh and och 3 groups with the hydroxyl groups.
In summary, two feruloyl esterases from aspergillus show 92. Pdf purification and characterization of a ferulic acid. One enzyme of that system is obtainable from aspergillus. Thus, the two substrates of this enzyme are feruloylpolysaccharide and h 2 o, whereas its two products are ferulate and polysaccharide. The gene product, faea, was overexpressed in wildtype a. Jul 01, 2006 read esterification of ferulic acid with polyols using a ferulic acid esterase from aspergillus niger, biochimica et biophysica acta bba general subjects on deepdyve, the largest online rental service for scholarly research with thousands of academic publications available at your fingertips. In this paper we show that the extent of the synergy between an aspergillus niger ferulic acid esterase faeiii and various xylanases.
After centrifugation 10,000 g, 15 min, the ferulic acid content of the supernatant was determined by hplc. Iucr structure of a feruloyl esterase from aspergillus niger. Regulation of the feruloyl esterase faea gene from. Highlevel production of recombinant aspergillus niger. As a component of the array of enzymes produced by microorganisms to deconstruct plant cell walls, feruloyl esterases hydrolyze phenolic groups involved in the crosslinking of arabinoxylan to other polymeric structures. The breakage of one or both ester bonds from dehydrodimer crosslinks between plant cell wall polymers is essential for optimal action of carbohydrases on these substrates. The protein has an alphabetahydrolase structure with a serhisasp catalytic triad.
A novel ferulic acid esterase gene from rumen fungus anaeromyces mucronatus was cloned, heteroexpressed in escherichia coli and characterized. Thermal denaturation was irreversible and the loss of activity was dependent on ph. Production of a chimeric enzyme tool associating the trichoderma reesei swollenin with the aspergillus niger feruloyl esterase a for release of ferulic acid. Muralikrishna, scientistf, department of biochemistry and nutrition.
Extracellular feruloyl esterases from the filamentous fungus aspergillus niger are induced by growth on oat spelt xylan osx, which contains no detectable esterified ferulic acid. Purification and characterization of a ferulic acid. Reducing cell wall feruloylation by expression of a fungal. Here, we describe the first crystal structure of the nonmodular typea feruloyl esterase from aspergillus niger anfaea solved at 2. Pdf purification and characterization of a ferulic acid esterase. Faulds and williamson 1995 reported that a maximum of 95% of total fa was proficiently released from wheat bran after a purified ferulic acid esterase fae from aspergillus niger and a trichoderma viride xylanase were incubated together for five hours.
A preparation of feruloyl esterase was obtained from post culture medium of aspergillus niger. A ferulic acid esterase faeiii, which was induced by growth of aspergillus niger cbs 120. The purification procedure utilized ammonium sulphate precipitation, hydrophobic interaction and anionexchange chromatography. The systematic name of this enzyme class is feruloyl esterase. If, faulds cb 2001 highlevel production of recombinant aspergillus niger cinnamoyl esterase. One unit of esterase activity was defined as the amount of enzyme required to release 1 mol hydroxycinnamic acidmin. The crystallographic structure of feruloyl esterase from aspergillus niger has been determined to a resolution of 1.
The structure of the enzymeproduct complex was determined to a resolution of 1. The apparent specific activity of ferulic acid esterases on cell wall polysaccharide materials is enhanced by the presence of xylanases. Background ferulic acid levels were subtracted during calculations. Finally, the swoifaea protein was tested for release of ferulic acid from wheat bran. Expression of the feruloyl esterase encoding gene faea from aspergillus niger depends on dxylose expression is mediated by xlnr, the xylanolytic transcriptional activator and on a second system that responds to aromatic compounds with a defined ring structure, such as ferulic acid and vanillic acid. The product of the hydrolysis, ferulic acid, can be used in the production of compounds of commercial interest, e. Released ferulic acid was further used for the production of the natural aroma vanillin. By day 5, residual levels of the free acid were below the limits of detection by hplc 4. The vanillin generating pathway extends from ferulic acid as precider, with isolation of aspergillus producing feruloyl esterase, which is characteristic of the enzyme. The objectives were to determine the effects of particle size. Oat hulls, an agricultural byproduct, contain a relatively high amount of ferulic acid fa. An aspergillus niger esterase ferulic acid esterase iii and. Abstract extracellular feruloyl esterases from the filamentous fungus aspergillus niger are induced by growth on oat spelt xylan osx, which. Influence of ferulic acid on the production of feruloyl.
Faeiii accounted for most of the feruloyl esterase activity. Moreover, the application of feruloyl esterase for beer production and biofuel production is explained. The initial rate of synthesis decreased by 1520% when the water content of the microemulsion was increased from 1. Release of ferulic acid from maize bran and derived. Ferulic acid fa is a component of plant cell walls that has applications in food, cosmetic, and health products, but its applications are limited by its high insolubility. Feruloyl esterase from aspergillus niger request pdf. Thus, the two substrates of this enzyme are feruloylpolysaccharide and h2o, whereas its two products are ferulate and polysaccharide. In this paper, aspergillus ferulic acid esterase fae was investigated for its ability to release fa from oat hulls. We report the cloning and characterization of a gene encoding a ferulic acid esterase, faea, from aspergillus niger and aspergillus tubingensis. Esterification of ferulic acid with polyols using a. An aspergillus niger esterase ferulic acid esterase iii and a recombinant pseudomonas fluorescens subsp. One, designated i, has a mr of 2,000, is probably dimeric, and has a pi of 3. Regulation of the feruloyl esterase faea gene from aspergillus niger.
A cinnamoyl esterase, ferulic acid esterase a, from aspergillus niger releases ferulic acid and 55 and 8o4dehydrodiferulic acids from plant cell walls. The stability of ferulic acid esterase iii faeiii from aspergillus niger was examined using chemical and thermal denaturation. Ferulic acid residues in plant cell walls are linked mainly to. The second, designated ii, was partially purified and is monomeric mr 29,000, with a pi of 3. Faea also hydrolyzed smaller amounts of ferulic acid from sugar beet pectin, but the rate was hardly affected by addition of an endopectin lyase. Anfaea displays an alphabeta hydrolase fold similar to that found in fungal lipases and different from that reported for other feruloyl esterases. Aug 15, 2005 in all measurements, the free acid present in samples pretreated with glacial acetic acid was subtracted from that in the test assays. We synthesized watersoluble fa derivatives by esterification of fa with diglycerol dg using feruloyl esterase purified from a commercial enzyme preparation produced by aspergillus niger. Purification and characterization of a ferulic acid esterase. Characterization of two distinct feruloyl esterases, aofaeb and aofaec, from aspergillus oryzae. One of the most thoroughly studied faes is feruloyl esterase a faea or faeiii from aspergillus niger. Release of ferulic acid from wheat bran by a ferulic acid esterase faeiii from aspergillus niger.
Ferulic acid was efficiently released from a wheat bran preparation by a ferulic acid esterase from aspergillus niger faeiii when incubated together wit faeiii by itself could release ferulic acid but at a level almost 24fold lower than that obtained in the presence of the xylanase 2 u. A cinnamoyl esterase from aspergillus nigercan break plant. Ferulic acid was very quickly lost from the medium during the first 24 h of growth. An aspergillus niger esterase ferulic acid esterase iii.
Enzymatic esterification of ferulic acid with glycerol and various sugars, which include arabinose, fructose, galactose, glucose, rhamnose, xylose, maltose, sucrose, and trehalose, was examined by incubating acceptors with faepl in the presence of ferulic acid at 50 c for 30 min at ph 4. Feruloyl esterase activity 1 mu was defined as the enzyme produced by 1 g of carbon source that released 1. We show that incubation of wheat and barley cell walls with ferulic acid esterase from aspergillus niger faeiii or pseudomonas fluorescens xy1d, together with either xylanase i from aspergillus niger. Pdf influence of ferulic acid on the production of. Chemical and thermal stability of ferulic acid esterase.
Structural comparison with a ferulic acid esterase fae from aspergillus niger indicated that al axea has a conserved catalytic machinery. I declare that the thesis entitled purification and biochemical characterization of ferulic acid esterase from aspergillus niger cfr 1105 submitted to the university of mysore, mysore for the award of the degree of doctor of philosophy in biochemistry is the result of the work carried out by me under the guidance of dr. Dec 31, 2002 read feruloyl esterase from aspergillus niger, process biochemistry on deepdyve, the largest online rental service for scholarly research with thousands of academic publications available at your fingertips. Ab we report the cloning and characterization of a gene encoding a ferulic acid esterase, faea, from aspergillus niger and aspergillus tubingensis. This is important for opening the cell wall structure, making material more accessible to glycosyl hydrolases. Characterization of a feruloyl esterase from aspergillus terreus facilitates the division of fungal enzymes from.