A novel Aspergillus oryzae diglycosidase that hydrolyzes 6-O-α-L-rhamnosyl-β-D-glucoside from flavonoids

Applied Microbiology and Biotechnology
Mai IshikawaTakuya Koseki

Abstract

α-L-Rhamnosyl-β-D-glucosidase (rutinosidase) hydrolyzes the glycosidic linkage between the disaccharide 6-O-α-L-rhamnosyl-β-D-glucoside (rutinose) and the aglycone. We identified a hypothetical protein (annotated as AO090012000917) encoded in the Aspergillus oryzae genome that exhibits sequence similarity with Aspergillus niger rutinosidase. The recombinant enzyme was expressed in Pichia pastoris GS115 and purified as a glyco-protein with apparent molecular mass of 65-75 kDa by SDS-PAGE. After N-deglycosylation, we observed a 42- and 40-kDa band, representing proteins before and after N-terminal signal peptide processing, respectively. Optimal enzymatic activity was observed at pH 4.0 and temperature of 45 °C. This enzyme is also significantly thermo-stable, with 90% activity retained after 1 h at 45 °C and 70% activity retained after 4 h, even at 50 °C. Biochemical characterization revealed that the enzyme has higher substrate specificity for 3-O-linked flavonoid β-rutinosides like rutin and kaempferol-3-O-rutinoside, than for 7-O-linked flavonoid β-rutinoside like hesperidin. However, no activity was found with naringin, diosmin, monoglycosylated chromogenic substrates, and polymeric laminarin substrate. Kinetic analyses show...Continue Reading

References

Oct 14, 1988·Biochemical and Biophysical Research Communications·K MizunoH Matsuo
Aug 3, 2000·Bioscience, Biotechnology, and Biochemistry·T NarikawaT Fujii
May 31, 2002·Bioscience, Biotechnology, and Biochemistry·Shigeru YamamotoKanzo Sakata
Dec 14, 2002·Plant Physiology·Masaharu MizutaniKanzo Sakata
Sep 28, 2004·Proceedings of the National Academy of Sciences of the United States of America·Mark A RobinsonBenjamin G Davis
Nov 29, 2005·Plant Physiology and Biochemistry : PPB·Fumi NakanishiKoichiro Shimomura
Mar 25, 2006·Bioscience, Biotechnology, and Biochemistry·Kazutaka TsuruhamiTaichi Usui
May 10, 2007·Molecular Biology and Evolution·Koichiro TamuraSudhir Kumar
Sep 21, 2012·BMC Evolutionary Biology·Henrik AspeborgBernard Henrissat
Sep 27, 2013·Journal of Agricultural and Food Chemistry·Shigeru KatayamaSoichiro Nakamura
Nov 26, 2013·Nucleic Acids Research·Vincent LombardBernard Henrissat
Apr 23, 2014·The Journal of Biological Chemistry·Hiromichi SainoMasaharu Mizutani
Jan 13, 2016·The Journal of Biological Chemistry·Tomohiko MatsuzawaKatsuro Yaoi

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Citations

Jul 25, 2018·International Journal of Molecular Sciences·Kristýna SlámováKateřina Valentová
Nov 11, 2019·Applied Microbiology and Biotechnology·Gisela WeizJavier D Breccia
Aug 18, 2018·Applied Microbiology and Biotechnology·Takuya KosekiYoshihito Shiono
Nov 22, 2020·Applied and Environmental Microbiology·Koki MakabeTakuya Koseki
Mar 22, 2021·Applied Microbiology and Biotechnology·Micaela BaglioniLaura S Mazzaferro
Aug 12, 2021·Journal of Bioscience and Bioengineering·Tomoe KatoTakuya Koseki

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Methods Mentioned

BETA
PCR
gel filtration
electrophoresis
Protein Assay

Software Mentioned

ClustalW
SignalP
Molecular Evolutionary Genetics Analysis
GraFit

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