title: Chemoenzymatic synthesis of α-L-rhamnosides using recombinant α-L-rhamnosidase from Aspergillus terreus.
abstract: This study describes an efficient, large scale fermentation of a recombinant α-L-rhamnosidase originating from Aspergillus terreus. High-cell-density Pichia pastoris fermentation resulted in yields up to 627 U/L/h. The recombinant enzyme was used for the reverse rhamnosylation of various small organic compounds. A full factorial experimental design setup was applied to identify the importance of temperature, substrate concentrations, solvent type and concentration as well as the acidity of the reaction mixture. Careful optimization of these parameters allowed the synthesis of a range of α-L-rhamnosides among which cyclohexyl α-L-rhamnopyranoside, anisyl α-L-rhamnopyranoside and 2-phenylethyl α-L-rhamnopyranoside. In addition, α-L-rhamnosylation of phenolic hydroxyls in phenols such as hydroquinone, resorcinol, catechol and phenol was observed, which is a rather unique reaction catalyzed by glycosidases.
authors.affiliation: Centre for Industrial Biotechnology and Biocatalysis, Department of Biochemical and Microbial Technology, Faculty of Biosciences Engineering, Ghent University, Coupure Links 653, B-9000 Ghent, Belgium. Electronic address: karel.dewinter@ugent.be.
authors.forename: Karel
authors.initials: K
authors.lastname: De Winter
authors.affiliation: Laboratory of Biotransformation, Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, CZ 142 20 Prague, Czech Republic. Electronic address: daniela.simcikova@gmail.com.
authors.forename: Daniela
authors.initials: D
authors.lastname: Šimčíková
authors.affiliation: Centre for Industrial Biotechnology and Biocatalysis, Department of Biochemical and Microbial Technology, Faculty of Biosciences Engineering, Ghent University, Coupure Links 653, B-9000 Ghent, Belgium. Electronic address: bram.schalck@ugent.be.
authors.forename: Bram
authors.initials: B
authors.lastname: Schalck
authors.affiliation: Laboratory of Biotransformation, Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, CZ 142 20 Prague, Czech Republic. Electronic address: lenka@wcontact.cz.
authors.forename: Lenka
authors.initials: L
authors.lastname: Weignerová
authors.affiliation: Laboratory of Molecular Structure Characterization, Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, CZ 142 20 Prague, Czech Republic. Electronic address: pelantova@biomed.cas.cz.
authors.forename: Helena
authors.initials: H
authors.lastname: Pelantova
authors.affiliation: Centre for Industrial Biotechnology and Biocatalysis, Department of Biochemical and Microbial Technology, Faculty of Biosciences Engineering, Ghent University, Coupure Links 653, B-9000 Ghent, Belgium. Electronic address: wim.soetaert@ugent.be.
authors.forename: Wim
authors.initials: W
authors.lastname: Soetaert
authors.affiliation: Centre for Industrial Biotechnology and Biocatalysis, Department of Biochemical and Microbial Technology, Faculty of Biosciences Engineering, Ghent University, Coupure Links 653, B-9000 Ghent, Belgium. Electronic address: tom.desmet@ugent.be.
authors.forename: Tom
authors.initials: T
authors.lastname: Desmet
authors.affiliation: Laboratory of Biotransformation, Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, CZ 142 20 Prague, Czech Republic. Electronic address: kren@biomed.cas.cz.
authors.forename: Vladimír
authors.initials: V
authors.lastname: Křen
mesh_terms: D001230:Aspergillus / Q000235:genetics / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000096:biosynthesis*; D011995:Recombination, Genetic
chemical_list: D006026:Glycoside Hydrolases; C022525:alpha-L-rhamnosidase
keywords: Aspergillus terreus; Glycosylation; Phenolic hydroxyl; Reverse hydrolysis; α-l-Rhamnosidase