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Discovery of solabiose phosphorylase and its application for enzymatic synthesis of solabiose from sucrose and lactose

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Title: Discovery of solabiose phosphorylase and its application for enzymatic synthesis of solabiose from sucrose and lactose
Authors: Saburi, Wataru Browse this author →KAKEN DB
Nihira, Takanori Browse this author
Nakai, Hiroyuki Browse this author
Kitaoka, Motomitsu Browse this author
Mori, Haruhide Browse this author →KAKEN DB
Issue Date: 2022
Publisher: Nature Portfolio
Journal Title: Scientific reports
Volume: 12
Issue: 1
Start Page: 259
Publisher DOI: 10.1038/s41598-021-04421-2
Abstract: Glycoside phosphorylases (GPs), which catalyze the reversible phosphorolysis of glycosides, are promising enzymes for the efficient production of glycosides. Various GPs with new catalytic activities are discovered from uncharacterized proteins phylogenetically distant from known enzymes in the past decade. In this study, we characterized Paenibacillus borealis PBOR_28850 protein, belonging to glycoside hydrolase family 94. Screening of acceptor substrates for reverse phosphorolysis, in which alpha -d-glucose 1-phosphate was used as the donor substrate, revealed that the recombinant PBOR_28850 produced in Escherichia coli specifically utilized d-galactose as an acceptor and produced solabiose (beta -d-Glcp-(1 -> 3)-d-Gal). This indicates that PBOR_28850 is a new GP, solabiose phosphorylase. PBOR_28850 catalyzed the phosphorolysis and synthesis of solabiose through a sequential bi-bi mechanism involving the formation of a ternary complex. The production of solabiose from lactose and sucrose has been established. Lactose was hydrolyzed to d-galactose and d-glucose by beta -galactosidase. Phosphorolysis of sucrose and synthesis of solabiose were then coupled by adding sucrose, sucrose phosphorylase, and PBOR_28850 to the reaction mixture. Using 210 mmol lactose and 280 mmol sucrose, 207 mmol of solabiose was produced. Yeast treatment degraded the remaining monosaccharides and sucrose without reducing solabiose. Solabiose with a purity of 93.7% was obtained without any chromatographic procedures.
Type: article
Appears in Collections:農学院・農学研究院 (Graduate School of Agriculture / Faculty of Agriculture) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

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