title: Overexpression of Aspergillus tubingensis faeA in protease-deficient Aspergillus niger enables ferulic acid production from plant material.
abstract: The production of ferulic acid esterase involved in the release of ferulic acid side groups from xylan was investigated in strains of Aspergillus tubingensis, Aspergillus carneus, Aspergillus niger and Rhizopus oryzae. The highest activity on triticale bran as sole carbon source was observed with the A. tubingensis T8.4 strain, which produced a type A ferulic acid esterase active against methyl p-coumarate, methyl ferulate and methyl sinapate. The activity of the A. tubingensis ferulic acid esterase (AtFAEA) was inhibited twofold by glucose and induced twofold in the presence of maize bran. An initial accumulation of endoglucanase was followed by the production of endoxylanase, suggesting a combined action with ferulic acid esterase on maize bran. A genomic copy of the A. tubingensis faeA gene was cloned and expressed in A. niger D15#26 under the control of the A. niger gpd promoter. The recombinant strain has reduced protease activity and does not acidify the media, therefore promoting high-level expression of recombinant enzymes. It produced 13.5 U/ml FAEA after 5 days on autoclaved maize bran as sole carbon source, which was threefold higher than for the A. tubingensis donor strain. The recombinant AtFAEA was able to extract 50 % of the available ferulic acid from non-pretreated maize bran, making this enzyme suitable for the biological production of ferulic acid from lignocellulosic plant material.
authors.affiliation: Department of Microbiology, Stellenbosch University, Private Bag X1, Matieland, 7602, South Africa.
authors.forename: Eunice N
authors.initials: EN
authors.lastname: Zwane
authors.forename: Shaunita H
authors.initials: SH
authors.lastname: Rose
authors.forename: Willem H
authors.initials: WH
authors.lastname: van Zyl
authors.forename: Karl
authors.initials: K
authors.lastname: Rumbold
authors.forename: Marinda
authors.initials: M
authors.lastname: Viljoen-Bloom
mesh_terms: D001230:Aspergillus / Q000201:enzymology* / Q000235:genetics; D001234:Aspergillus niger / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D002265:Carboxylic Ester Hydrolases / Q000235:genetics / Q000378:metabolism*; D002480:Cellulase / Q000378:metabolism; D003373:Coumaric Acids / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000378:metabolism; D003313:Zea mays / Q000378:metabolism
chemical_list: D003373:Coumaric Acids; C004999:ferulic acid; D002265:Carboxylic Ester Hydrolases; C070196:feruloyl esterase; D002480:Cellulase; D010447:Peptide Hydrolases