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Biotechnology and Biological Transformations
Novel #-1,3/#-1,4-glucosidase from Aspergillus niger exhibits unique transglucosylation to generate high levels of nigerose and kojibiose Min Ma, Masayuki Okuyama, Takayoshi Tagami, Asako Kikuchi, Patcharapa Klahan, and Atsuo Kimura J. Agric. Food Chem., Just Accepted Manuscript • DOI: 10.1021/acs.jafc.8b07087 • Publication Date (Web): 26 Feb 2019 Downloaded from http://pubs.acs.org on February 27, 2019
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Journal of Agricultural and Food Chemistry
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Novel
α-1,3/α-1,4-glucosidase
from
Aspergillus
niger
exhibits
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transglucosylation to generate high levels of nigerose and kojibiose
unique
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Min Ma, Masayuki Okuyama,* Takayoshi Tagami, Asako Kikuchi, Patcharapa
5
Klahan, and Atsuo Kimura*
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Research Faculty of Agriculture, Hokkaido University, Kita-9 Nishi-9, Kita-ku,
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Sapporo 060-8589, Japan
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*Correspondence
should
be
11
[email protected])
12
[email protected]).
addressed or
to:
Atsuo
Masayuki
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Tel: 81-11-706-2808, Fax: 81-11-706-2808.
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1 ACS Paragon Plus Environment
Kimura
Okuyama
(E-mail: (E-mail:
Journal of Agricultural and Food Chemistry
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Abstract
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α-Glucosidase from Aspergillus niger (AgdA; typical α-1,4-glucosidase) is known to
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industrially produce α-(1→6)-glucooligosaccharides. This fungus also has another α-
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glucosidase-like protein, AgdB. To learn its function, wild-type AgdB was expressed
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in Pichia pastoris. However, the enzyme displayed two electrophoretic forms due to
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heterogeneity of N-glycosylation at Asn354. The deglycosylation-mutant N354D
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shared the same properties with wild-type AgdB. N354D demonstrated hydrolytic
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specificity toward α-(1→3)- and α-(1→4)-glucosidic linkages, indicating that AgdB
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is an α-1,3/α-1,4-glucosidase. N354D-catalyzed transglucosylation from maltose was
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analyzed in short- and long-term reactions, enabling to learn the transglucosylation
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specificity and product accumulation, respectively. Short-term reaction (
