Heterologous expression of two Aspergillus niger feruloyl esterases in Trichoderma reesei for the production of ferulic acid from wheat bran
Feruloyl esterase (FAE)-encoding genes AnfaeA and AnfaeB were isolated from Aspergillus niger 0913. For overexpression of the two genes in Trichoderma reesei, constitutive and inductive expression plasmids were constructed based on parental plasmid pAg1-H3. The constructed plasmids contained AnfaeA or AnfaeB gene under the control of glyceraldehyde-3-phosphate dehydrogenase A gene (gpdA) promoter (from A. nidulans) or cellobiohydrolases I (cbh I) gene promoter (from T. reesei), and cbh I terminator from T. reesei. The target plasmids were transferred into T. reesei D-86271 (Rut-C30) by Agrobacterium tumefaciens-mediated transformation (ATMT), respectively. A high level of feruloyl esterase was produced by the recombinant fungal strains under solid-state fermentation, and the cbh I promoter was more efficient than the gpdA promoter in the expression of AnfaeA. The optimum temperatures and pH values were 50 °C and 5.0 for AnFAEA, and 35 °C and 6.0 for AnFAEB. The maximum production levels were 20.69 U/gsd for AnFAEA and 15.08 U/gsd for AnFAEB. The recombinant fungal enzyme systems could release 62.9% (for AnFAEA) and 52.2% (for AnFAEB) of total ferulic acids from de-starched wheat bran, which was higher than the 46.3% releasing efficiency of A. niger 0913. The supplement of xylanase from T. longibrachiatum in the enzymatic hydrolysis led to a small increment of the ferulic acids release.
KeywordsFeruloyl esterase genes Solid-state fermentation Trichoderma reesei Ferulic acid production
We are grateful to Prof. Gang Liu (Institute of Microbiology, CAS) for providing the plasmid pAg1-H3 and Agrobacterium tumefaciens AGL-1. This work was supported by grants from a 948 Research Project (No. 2013-4-16) from the State Forestry Administration of China, the Natural Science Fund for Provincial Colleges and University of Jiangsu Province, China (15KJB220003), the Research Fund for the Advanced Talents, Nanjing Forestry University (GXL201311) and the Priority Academic Program Development of Jiangsu Higher Education Institutions.
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Conflict of interest
The authors declare that they have no conflict of interest.
- 21.Mandels M, Andreotti RE (1978) Problems and challenges in the cellulose to cellulase fermentation. Process Biochem 13:6–13Google Scholar
- 23.Khang CH, Park SY, Rho HS, Lee YH, Kang S (2006) Filamentous fungi (Magnaporthe grisea and Fusarium oxysporum). Methods Mol Biol 344:403–420. Agrobacterium protocols, 2/e, volume 2. Edited by Wang K, Humana, TotowaGoogle Scholar
- 32.Levasseur A, Saloheimo M, Navarro D, Andberg M, Monot F, Nakari-Setala T, Asther M, Record E (2006) Production of a chimeric enzyme tool associating the Trichoderma reesei swollenin with the Aspergillus niger feruloyl esterase A for release of ferulic acid. Appl Microbiol Biot 73:872–880CrossRefGoogle Scholar
- 34.Kroon PA, Faulds CB, Williamson G (1996) Purification and characterization of a novel esterase induced by growth of Aspergillus niger on sugar-beet pulp. Biotechnol Appl Biochem 23:255–262Google Scholar
- 36.de Vries RP, Michelsen B, Poulsen CH, Kroon PA, van den Heuvel RH, Faulds CB, Williamson G, van den Hombergh JP, Visser J (1997) The faeA genes from Aspergillus niger and Aspergillus tubingensis encode ferulic acid esterases involved in degradation of complex cell wall polysaccharides. Appl Environ Microbiol 63:4638–4644Google Scholar
- 37.Bartolomé B, Faulds CB, Tuohy M, Hazlewood GP, Gilbert HJ, Williamson G (1995) Influence of different xylanases on the activity of ferulic acid esterase on wheat bran. Biotechnol Appl Biochem 22:65–73Google Scholar
- 38.de Vries RP, Visser J (1999) Regulation of the feruloyl esterase (faeA) gene from Aspergillus niger. Appl Environ Microbiol 65:5500–5503Google Scholar