Liquid-Phase and Ultrahigh-Frequency-Acoustofluidics-Based Solid-Phase Synthesis of Biotin-Tagged 6′/3′-Sialyl-N-Acetylglucosamine by Sequential One-Pot Multienzyme System

Yuguang Du,Tiechuan Li,Hongzhi Cao,Lishi Ren,Meishan Pei,Lina Wu,Jian-Jun Li,Xuexin Duan,Zhenxing Zhang,Mengge Gong

doi:10.3390/catal10111347

Abstract

6′/3′-Sialylated N-acetyllactosamine (6′/3′-SLN) is important for discrimination of the source (human or avian) of influenza virus strains. Biotinylated oligosaccharides have been widely used for analysis and quick detection. The development of efficient strategies to synthesize biotin-tagged 6′/3′-SLN have become necessary. Effective mixing is essential for enzymatic solid-phase oligosaccharide synthesis (SPOS). In the current study, newly developed technology ultrahigh-frequency-acoustofluidics (UHFA), which can provide a powerful source for efficient microfluidic mixing, solid-phase oligosaccharide synthesis and one-pot multienzyme (OPME) system, were used to develop a new strategy for oligosaccharide synthesis. Firstly, biotinylated N-acetylglucosamine was designed and chemically synthesized through traditional approaches. Secondly, biotinylated 6′- and 3′-sialyl-N-acetylglucosamines were prepared in solution through two sequential OPME modules in with a yield of ~95%. Thirdly, 6′-SLN was also prepared through UHFA-based enzymatic solid-phase synthesis on magnetic beads with a yield of 64.4%. The current strategy would be potentially used for synthesis of functional oligosaccharides.

Highlights

Proteins, nucleic acids, lipids, and carbohydrates are four main types of biomolecules forming the basis of life
The starting biotinylated monosaccharide acceptor-biotin-tagged N-acetylglucosamine was designed and synthesized through well-known procedures shown in Scheme 1 (Supplementary materials)
Compound 2 was prepared from compound with large excess of NaN3 at 95% yield

Summary

Introduction

Nucleic acids, lipids, and carbohydrates are four main types of biomolecules forming the basis of life. Nucleic acids and lipids, carbohydrates play key roles in many biological processes such as protein conformation [1], molecular recognition [2], cell proliferation and differentiation [3], and are closely related with occurrence and development of many diseases [4]. Compared with nucleic acids and proteins, studies toward carbohydrates are lagging behind. Sialic acid is generally located at the nonreducing ends of the sugar chains [7]. This outmost position and ubiquitous distribution enable sialylated glycans to be involved in numerous cellular processes. Studies have shown that hemagglutinin (HA) of human influenza virus strains preferentially binds to oligosaccharides that terminate with

Methods

Results

Conclusion