Heterologous expression of two Aspergillus niger feruloyl esterases in Trichoderma reesei for the production of ferulic acid from wheat bran

Abstract

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.

Characterization and functional analysis of two novel thermotolerant α-l-arabinofuranosidases belonging to glycoside hydrolase family 51 from Thielavia terrestris and family 62 from Eupenicillium parvum

“…To facilitate purification, 6× his tag was included at the C-terminal of the expressed protein. To express the gene in T. reesei , the gene fragment without the signal sequence was PCR amplified using primers Abf51A_f2 and Abf51A_r2 (Supplementary Table S1 ) and the gene fragment was ligated into plasmid pAg-PTcbh1 (Long et al 2018c ) by Hieff Clone™ One Step PCR Cloning Kit (Yeasen, Shanghai, China). The recombinant plasmid was introduced into T. reesei strain D-86271 by Agrobacterium tumefaciens– mediated transformation method (Long et al 2018a ; Mullins et al 2001 ).…”

Isolation, identification and characterization of soil bacteria for the production of ferulic acid through co-culture fermentation using banana stem waste

“…FAE acts synergistically with other hemicellulases such as xylanase to maximize the microbial degradation of plant cell wall [9]. This has been proven by Uraji et al [10] in their finding where combinational enzymes have significantly enhanced FA production in rice bran, wheat bran and corncob.…”

Application of Michaelis–Menten in the kinetics of oil palm frond enzymatic hydrolysis for ferulic acid production

“…Hydrolysis process has been reported to be able to break these bonds and producing FA [17]. There are also reports that enzymatic hydrolysis with the presence of feruloyl esterase (FAE) have increased FA production due to the mechanism of FAE which cleave the hemicellulose in plant structure [9,18,19]. These reports indicate that the presence of FAE may induce FA production via enzymatic hydrolysis.…”