Biosynthesis of xylo-oligosaccharides from wheat straw xylan via the synergistic hydrolysis by xylanase Xyn11A and arabinofuranosidase Abf62A Synthetic Biology Journal    DOI: 10.12211/2096-8280.2025-037

Fig. 1 Cloning and expression analysis of Abf62A in P. pastoris

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The arabinofuranosidase gene Abf62A was successfully cloned from Aspergillus usamii and expressed in P. pastoris ［Fig. 1（a） and （b）］. Electrophoretic analysis confirmed the removal of the native secretion signal， yielding a mature 918-bp coding sequence ［Fig. 1（c）］. Homologous recombination generated the 3952-bp plasmid pGAPZαA-Abf62A， which was subsequently transformed into P. pastoris X33. SDS-PAGE of culture supernatants revealed distinct ～33 kDa protein bands ［Fig. 1（d）］， consistent with Abf62A’s predicted molecular mass （32.8 kDa）. Optimum enzyme production occurred at 30 °C and pH 5.5 with 0.5% methanol induction.

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• Fig. 2  Abf62A enzyme activity of different strains
• Fig. 3  Effects of culture time (a) and temperature (b) on the enzyme activity of Abf62A fermentation liquor.
• Fig. 4  SDS-PAGE results Abf62A protein(Lane A: Abf62A fermentation liquor; Lane B: Abf62A flow-through solution; Lane C: Abf62A eluate solution; Lane D: Abf62A pure enzyme solution)
• Fig. 5  The optimum pH and pH stability of Abf62A (a) and its optimum temperature and temperature stability (b)
• Fig. 6  Integrated process for valorizing wheat straw into antioxidant xylooligosaccharides
• Table 1  Analysis of Xyn11A enzymatic hydrolysis products of different WS xylan
• Table 2  Sugar content and proportion in xylan hydrolysates of different WS.
• Fig. 7  Radical scavenging activity of xylo-oligosaccharides (XOS)(Data represent mean values of triplicate measurements.)