Toward Automated Enzymatic Synthesis of Oligosaccharides

Abstract

Oligosaccharides together with oligonucleotides and oligopeptides comprise the three major classes of natural biopolymers. Automated systems for oligonucleotide and oligopeptide synthesis have significantly advanced developments in biological science by allowing nonspecialists to rapidly and easily access these biopolymers. Researchers have endeavored for decades to develop a comparable general automated system to synthesize oligosaccharides. Such a system would have a revolutionary impact on the understanding of the roles of glycans in biological systems. The main challenge to achieving automated synthesis is the lack of general synthetic methods for routine synthesis of glycans. Currently, the two main methods to access homogeneous glycans and glycoconjugates are chemical synthesis and enzymatic synthesis. Enzymatic glycosylation can proceed stereo- and regiospecifically without protecting group manipulations. Moreover, the reaction conditions of enzyme-catalyzed glycosylations are extremely mild when compared to chemical glycosylations. Over the past few years methodology toward the automated chemical synthesis of oligosaccharides has been developed. Conversely, while automated enzymatic synthesis is conceptually possible, it is not as well developed. The goal of this survey is to provide a foundation on which continued technological advancements can be made to promote the automated enzymatic synthesis of oligosaccharides.

Machine‐Driven Enzymatic Oligosaccharide Synthesis by Using a Peptide Synthesizer

“…At the same time, many works found that enzymatic glycosylations can be performed on solution-phase resins/polymers. [20] However, the use of solution-phase supports or polymers increases the difficulty of product purification. In 2010, Nishimura and co-workers used a globular protein-like PAMAM dendrimer for automated synthesis, which allows the product to be separated by ultrafiltration.…”

DNA Encoded Glycan Libraries as a next-generation tool for the study of glycan-protein interactions

“…[26] Song's work established the feasibility of DNA encoding and NGS in multiplex glycan interaction studies, while the glycan-DNA conjugation method used may limit the accessibility to limited glycan content and thereby compromise the true potential DNA encoding. Advances in the synthesis of carbohydrates via organic, enzymatic, chemoenzymatic and machine-driven synthesis [27] have potential to eliminate the bottleneck in glycan interaction studies, i.e., availability of glycan probes. These new developments necessitate novel approaches for high-content and high-throughput methods for glycan interaction studies.…”

Reengineering Chemical Glycosylation: Direct, Metal‐Free Anomeric O‐Arylation of Unactivated Carbohydrates

“…Ad issociative mechanistic spectrum spanning S N 1a nd S N 2i sf requently operational therebyr educing the efficiency. An amalgamation of regio-and stereoselectivity complications, further compounded by complex conformational equilibria, renders carbohydrate coupling protocols less general than the aforementioned biomolecular classes.A lthough advances in automation have significantly alleviated the synthesis bottleneck, [5] innovative coupling methods to construct the acetal junction intrinsict oc arbohydrates must be developedi np arallel. The use of stable aryliodonium salts facilitates af ormalO ÀHf unctionalisation reaction.…”

“…[2] This is particularly prominent in phenolicg lycosides such as the antibiotic Vancomycin. An amalgamation of regio-and stereoselectivity complications, further compounded by complex conformational equilibria, renders carbohydrate coupling protocols less general than the aforementioned biomolecular classes.A lthough advances in automation have significantly alleviated the synthesis bottleneck, [5] innovative coupling methods to construct the acetal junction intrinsict oc arbohydrates must be developedi np arallel. [4] In contrastt op eptides and nucleic acids, the efficiency with whichc omplex sugars are produced biosynthetically cannote asily be emulated in al aboratory paradigm.…”

“…[3] However,e xploring and harnessing carbohydrate functioni sf requently compromised by synthetic challenges. An amalgamation of regio-and stereoselectivity complications, further compounded by complex conformational equilibria, renders carbohydrate coupling protocols less general than the aforementioned biomolecular classes.A lthough advances in automation have significantly alleviated the synthesis bottleneck, [5] innovative coupling methods to construct the acetal junction intrinsict oc arbohydrates must be developedi np arallel. An amalgamation of regio-and stereoselectivity complications, further compounded by complex conformational equilibria, renders carbohydrate coupling protocols less general than the aforementioned biomolecular classes.A lthough advances in automation have significantly alleviated the synthesis bottleneck, [5] innovative coupling methods to construct the acetal junction intrinsict oc arbohydrates must be developedi np arallel.…”