Release of Arabinose from Wheat Insoluble Arabinoxylan by the Action of α-L-Arabinofuranosidase in Synergism with Endo-Xylanases

Wheat arabinoxylan (water-insoluble fraction) contains ~36% arabinose which may include both singly or doubly substitutions at C2/C3 of the Xylp units. α-L-Arabinofuranosidses (ABFs) of two GH families were analyzed for their respective activities on the hydrolysis of Xylp-Araf. BaABF (GH43) produced twice the yield of arabinose residues from the heteroxylan compared to AnABF (GH51) under the same reaction conditions. Two endo-xylanases (of GH10 and 11) also showed differential hydrolytic activities on the Xylp chain, with the GH10 XYN-ATM double the amount of reducing sugar yield (as xylose equivalent) than using the GH11 XYN-M3. When the ABF and XYN were combined in optimial ratios, a synergistic increase of 73.8% in arabinose yield was observed.

Cloning of an α-L-Arabinofuranosidase and Characterization of Its Action on Mono- and Di-Substituted Xylopyranosyl Units

An α-L-arabinofuranosidase (ARF) gene of 1503 bp was synthesized, subcloned into pET26b vector, and expressed in Escherichia coli. The enzyme was purified in active form, and consisted of 500 amino acid residues, corresponding to 55 kD based on SDS-PAGE. The affinity-purified protein was characterized using arabinofuranosyl xylooligosaccharides (AXOS) as substrates. The pH effect was investigated showing an optimum at pH 5.5. XaARF catalyzed the cleavage of arabinose at C3 of the xylopyranosyl unit efficiently if the arabinofuranosyl substitution was at the terminal compared to internal xylose units. The enzyme was able to act on di-substituted xylopyranosyl units with the first cleavage at C3 followed by C2 linkages.

微生物来源的α-L-阿拉伯呋喃糖苷酶的研究进展

α-L-阿拉伯呋喃糖苷酶(Alpha-L-arabinofuranosidase,α-L-AFase)属于糖苷水解酶类,主要功能是水解阿拉伯木聚糖中的阿拉伯糖取代基,并与其他水解酶协同促进半纤维素的降解,从而改善半纤维素的生物转化率。本文综述了近年来微生物来源α-L-AFase的酶学特性、性质改良、结构、催化机制和协同作用效应等方面的研究进展,发现不同微生物来源的α-L-AFase的理化性质、分子结构和催化机制均存在着多样性,重组表达的方法能有效改善α-L-AFase的活性和稳定性,且α-L-AFase与其他半纤维素水解酶的协同催化作用使底物的转化效率显著提高。α-L-AFase广泛应用于改善面团发酵质地、果汁澄清、酿酒工艺优化、制备高消化率饲料以及功能性保健产品研制等多个领域。本文为后续α-L-AFase的基础研究、开发和利用提供一定的参考。