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Enzymatic Glycosylation of Small Molecules University of Groningen Enzymatic Glycosylation of Small Molecules Desmet, Tom; Soetaert, Wim; Bojarova, Pavla; Kren, Vladimir; Dijkhuizen, Lubbert; Eastwick- Field, Vanessa; Schiller, Alexander; Křen, Vladimir Published in: Chemistry : a European Journal DOI: 10.1002/chem.201103069 IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite from it. Please check the document version below. Document Version Publisher's PDF, also known as Version of record Publication date: 2012 Link to publication in University of Groningen/UMCG research database Citation for published version (APA): Desmet, T., Soetaert, W., Bojarova, P., Kren, V., Dijkhuizen, L., Eastwick-Field, V., Schiller, A., & Křen, V. (2012). Enzymatic Glycosylation of Small Molecules: Challenging Substrates Require Tailored Catalysts. Chemistry : a European Journal, 18(35), 10786-10801. https://doi.org/10.1002/chem.201103069 Copyright Other than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons). Take-down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons the number of authors shown on this cover page is limited to 10 maximum. Download date: 24-09-2021 DOI: 10.1002/chem.201103069 Enzymatic Glycosylation of Small Molecules: Challenging Substrates Require Tailored Catalysts Tom Desmet,[b] Wim Soetaert,[b, c] Pavla Bojarov,[d] Vladimir Krˇen,[d] Lubbert Dijkhuizen,[e] Vanessa Eastwick-Field,[f] and Alexander Schiller*[a] 10786 2012 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Chem. Eur. J. 2012, 18, 10786 – 10801 REVIEW Abstract: Glycosylation can significantly improve the recent examples. Progress in the field of enzyme engineer- physicochemical and biological properties of small mole- ing and screening are expected to result in new applica- cules like vitamins, antibiotics, flavors, and fragrances. The tions of biocatalytic glycosylation reactions in various in- chemical synthesis of glycosides is, however, far from trivi- dustrial sectors. al and involves multistep routes that generate lots of waste. In this review, biocatalytic alternatives are present- Keywords: acceptor specificity · enzyme engineering · ed that offer both stricter specificities and higher yields. glycosylation · glycosyltransferase · high-throughput The advantages and disadvantages of different enzyme screening classes are discussed and illustrated with a number of Introduction cirrhizin, a terpenoid glycoside from sweetwood (Glycirhyza glabra) that loses most of its sweetness upon hydrolysis.[4] Besides being a source of energy and a structural compo- Since the transfer of a glycosyl group can influence both nent of the cell wall, carbohydrates also mediate various rec- the physicochemical and biological properties of an organic ognition processes when attached to proteins or lipids.[1] molecule, such processes may be used for a wide range of Well-known carbohydrate motifs of such glycoconjugates applications. The most obvious advantage of introducing a are the cancer epitope Sialyl Lewis X and the AB0 blood carbohydrate moiety is the increased solubility of hydropho- group determinants. In addition, glycosylation is an impor- bic compounds. This is nicely illustrated in flavonoids, the tant source of structural diversity of natural products, such pharmaceutical properties of which can often be efficiently as alkaloids, steroids, flavonoids, and antibiotics. Glycosides exploited only in the form of their hydrophilic glycosyl de- typically display properties that differ from those of their rivatives.[2] Glycosylation may also be used to improve the non-glycosylated aglycons.[2] A prime example is naringin, a stability of labile molecules. A famous example is ascorbic flavanone glycoside that is responsible for the bitter taste of acid, a very sensitive vitamin, the long-term storage of citrus fruits. Removal of the glycon part eliminates the which can be drastically extended by glycosylation, resulting bitter taste, which is one of the main goals of enzymatic in high-value applications in cosmetics and tissue culturing.[5] treatment of grapefruit juice.[3] The opposite is true for gly- Another important application of glycosylation is the reduc- tion of skin irritation caused by hydroquinone, employed in cosmetics for its skin whitening effect.[6] Glycosides of fla- vors and fragrances, in turn, can function as controlled re- [a] Prof. A. Schiller lease compounds. The a-glucoside of l-menthol, for exam- Friedrich-Schiller-University Jena ple, is only slowly hydrolyzed in the mouth, resulting in a Institute for Inorganic and Analytical Chemistry [7] Humboldtstr. 8, 07743 Jena (Germany) prolonged sensation of freshness. Last but not least, it has E-mail: [email protected] been possible to modulate the activity spectrum of glyco- [b] Prof. T. Desmet, Prof. W. Soetaert peptide antibiotics by varying their carbohydrate moiety, in University of Ghent a process known as “glycorandomization”.[8,9] Centre for Industrial Biotechnology and Biocatalysis In view of these examples, the development of cheap and Coupure links 653, 9000 Gent (Belgium) efficient glycosylation technologies, useful both in the labo- [c] Prof. W. Soetaert ratory and in industry, is highly desirable. In this review, the BioBase Europe Pilot Plant Rodenhuizekaai 1, Havennummer 4200 challenges and recent innovations concerning the glycosyla- 9042 Gent (Belgium) tion of small, non-carbohydrate molecules are covered. [d] Dr. P. Bojarov, Prof. V. Krˇen Institute of Microbiology Academy of Sciences of the Czech Republic Chemical versus Enzymatic Glycosylation Vdenˇ sk 1083, 142 20 Prague (Czech Republic) [e] Prof. L. Dijkhuizen Microbiology, University of Groningen Glycosylation reactions by conventional chemical synthesis Groningen Biomolecular Sciences and are used intensively in the field of glycochemistry. Despite Biotechnology Institute (GBB) the variety of glycosylation protocols developed to date,[10–25] Nijenborgh 7 P.O. Box 11103 synthesis of glycosylated compounds largely relies on four 9700 CC Groningen (The Netherlands) non-enzymatic reactions (Scheme 1). One of the first was fa- [f] Dr. V. Eastwick-Field mously developed by Koenigs and Knorr, in which glycosyl Carbosynth Limited 8 & 9 Old Station Business Park halides, activated with silver salts, are used as glycosyl Compton, Newbury, Berkshire RG20 6NE (UK) donors.[26–28] Glycosyl trichloroacetimidates were later found Chem. Eur. J. 2012, 18, 10786 – 10801 2012 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.chemeurj.org 10787 A. Schiller et al. to be very powerful donor substrates with an excellent leav- activated by electrophilic reagents.[30] There are, however, ing group.[29] Alternatively, more stable glycosides, such as two major issues connected with the outcome of these reac- thioglycosides and n-pentenyl glycosides, can be used when tions, that is, the regioselectivity and the configuration of the glycosidic linkage. The former can be solved by appro- priate protection strategies, whereas the latter is strongly de- Tom Desmet received a Ph.D. in Biochem- pendent on the neighboring group participation of the C2 istry from Ghent University for work on substituent. Additionally, solvents and catalysts have an im- the structure–function relationships in portant effect on the anomeric outcome of glycosylation re- (hemi)cellulases. He is particularly interest- actions. ed in the engineering of carbohydrate- The chemical methods suffer from a number of draw- active enzymes for use in biocatalytic proc- esses, and has coordinated several projects backs: labor-intensive activation and protection procedures, in that field. After a postdoctoral stay at multistep synthetic routes with low overall yields, the use of Wageningen University, he was appointed toxic catalysts and solvents and the amount of waste.[31] To Associate Professor at the Centre for In- overcome these limitations, specific enzymes may be used dustrial Biotechnology and Biocatalysis, where he leads a team of about ten re- for the synthesis of glycosides. DeRoode et al. have calculat- searchers. Wim Soetaert holds a Ph.D. in Applied Bi- ological Sciences from Ghent University. Lubbert Dijkhuizen is Professor of Micro- After working in the starch industry for biology, University of Groningen. He is twelve years, he returned to the university scientific director of the Carbohydrate to become Associate Professor for Indus- Competence Center, a public-private part- trial Biotechnology. His research interests nership with 19 companies and 6 knowl- comprise the enzymatic and microbial con- edge institutes (http://www.cccresearch.nl). version of carbohydrates for the produc- His research focuses on the characteriza- tion of added-value chemicals. Wim Soe- tion and engineering of sterol/steroid con- taert also is the director of the Bio Base verting enzymes (Rhodococcus/Mycobac- Europe Pilot Plant as well as the founder terium), and starch and sucrose acting en- and chairman of Ghent Bio-Energy Valley. zymes (bacilli/lactobacilli). He is currently involved in EU FP7 research projects NO- VOSIDES and AMYLOMICS. Dr.
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