Heterologous expression and functional characterization of a GH10 endoxylanase from Aspergillus fumigatus var. niveus with potential biotechnological application

Abstract

Graphical abstract

A Fungal Versatile GH10 Endoxylanase and Its Glycosynthase Variant: Synthesis of Xylooligosaccharides and Glycosides of Bioactive Phenolic Compounds

“…This result indicates that this CBM1 domain is The maximum endoxylanase activity against AZCL-beechwood as substrate was observed at pH 5 and 70 • C (Figure S3). These values are comparable to those described for other fungal GH10 endoxylanases, ranging between 60 and 80 • C for the optimal temperature and pH 4.5 and 6 for the optimal pH [32][33][34][35][36][37].…”

“…This small preference for branched over linear xylans has also been reported for the GH10 endoxylanase XynD from Penicillium funiculosum [34]. rXynSOS was also able to hydrolyze CMC, although much less efficiently than the xylan substrates, as it was described with the GH10 endoxylanase AFUMN-GH10 from Aspergillus fumigatus [32]. Nevertheless, the enzyme could not hydrolyze microcrystalline cellulose and cellobiose, which are other specific substrates for enzymes with cellulolytic activity.…”

“…The kinetic parameters calculated for the best rXynSOS substrates, as well as for CMC as a representative of cellulose activity, are displayed in Table 2. The enzyme showed a high affinity for beechwood xylan and wheat arabinoxylan, with K m values of 1.07 and 1.94 g L −1 , respectively, which are better than the ones reported for most fungal GH10 endoxylanases [32,34,36,37]. The catalytic efficiencies (k cat /K m ) of rXynSOS on beechwood xylan (134.69 s −1 g −1 L) and wheat arabinoxylan (84.65 s −1 g −1 L) are in the same range as those of the GH10 endoxylanases Xyn10A from Penicillium oxalicum [36], PspXyn10 from Penicillum sp.…”

Combining pieces: a thorough analysis of light activation boosting power and co-substrate preferences for the catalytic efficiency of lytic polysaccharide monooxygenase MtLPMO9A

“…The coding sequence of MtLPMO9A from T. thermophilus M77 (MYCTH_85556, UniProt: KP901251) was amplified from genomic DNA, including the native signal peptide using 5'-agcatcattacacctcagcaATGCTGACAACAACCTTCGC-3'(forward) and 5'-taaatcactagatatctctaTTAGCAACGGAAGACAGCCG-3'(reverse) primers, and cloned into the pEXPYR vector (Gibson et al, 2009; Velasco et al, 2019) using Ligation-Independent Cloning (Camilo and Polikarpov, 2014). The expression plasmid was transformed into Aspergillus nidulans A773 (pyrG89; wA3; pyroA4), and the recombinant strain was selected using the pyrG auxotrophic maker as described earlier (Velasco et al, 2019).…”

Study of the lytic polysaccharide monooxygenases >i<Mt>/i<LPMO9A for biotechnological applications

“…The coding sequence of MtLPMO9A (MYCTH_85556, UniProt: KP901251) was amplified from genomic DNA, cloned, and the expression plasmid was transformed into Aspergilus nidulans A773 by Prof. Dr. Fernando Segato as previously described [51][52][53][54] and recombinants strains were kindly provided to Molecular Biotechnology Group.…”