Analysis of Transglucosylation Products of Aspergillus niger α-Glucosidase that Catalyzes the Formation of α-1,2- and α-1,3-Linked Oligosaccharides |
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Authors: | Atsushi Kawano Kansuke Fukui Yuji Matsumoto Atsushi Terada Akihiro Tominaga Nozomi Nikaido Takashi Tonozuka Kazuhide Totani Nozomu Yasutake |
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Affiliation: | 1. RD&E Center, Showa Sangyo Co., Ltd.;2. Division of Chemical Engineering and Biotechnology, Department of Engineering for Future Innovation, National Institute of Technology, Ichinoseki College;3. Department of Applied Biological Science, Tokyo University of Agriculture and Technology |
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Abstract: | According to whole-genome sequencing, Aspergillus niger produces multiple enzymes of glycoside hydrolases (GH) 31. Here we focus on a GH31 α-glucosidase, AgdB, from A. niger . AgdB has also previously been reported as being expressed in the yeast species, Pichia pastoris ; while the recombinant enzyme (rAgdB) has been shown to catalyze tranglycosylation via a complex mechanism. We constructed an expression system for A. niger AgdB using Aspergillus nidulans . To better elucidate the complicated mechanism employed by AgdB for transglucosylation, we also established a method to quantify glucosidic linkages in the transglucosylation products using 2D NMR spectroscopy. Results from the enzyme activity analysis indicated that the optimum temperature was 65 °C and optimum pH range was 6.0–7.0. Further, the NMR results showed that when maltose or maltopentaose served as the substrate, α-1,2-, α-1,3-, and small amount of α-1,1-β-linked oligosaccharides are present throughout the transglucosylation products of AgdB. These results suggest that AgdB is an α-glucosidase that serves as a transglucosylase capable of effectively producing oligosaccharides with α-1,2-, α-1,3-glucosidic linkages. |
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Keywords: | Aspergillus α -glucosidase glycoside hydrolase family 31 transglucosylation |
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