Liquid-Phase and Ultrahigh-Frequency-Acoustofluidics-Based Solid-Phase Synthesis of Biotin-Tagged 6′/3′-Sialyl-<i>N</i>-Acetylglucosamine by Sequential One-Pot Multienzyme System
Mengge Gong,
Tiechuan Li,
Lina Wu,
Zhenxing Zhang,
Lishi Ren,
Xuexin Duan,
Hongzhi Cao,
Meishan Pei,
Jian-Jun Li,
Yuguang Du
Affiliations
Mengge Gong
School of Chemistry and Chemical Engineering, University of Jinan, Jinan 250022, China
Tiechuan Li
State Key Laboratory of Precision Measuring Technology and Instruments, Tianjin University, Tianjin 300072, China
Lina Wu
National Key Laboratory of Biochemical Engineering, National Engineering Research Center for Biotechnology (Beijing), Key Laboratory of Biopharmaceutical Production & Formulation Engineering, PLA, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China
Zhenxing Zhang
Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China
Lishi Ren
National Key Laboratory of Biochemical Engineering, National Engineering Research Center for Biotechnology (Beijing), Key Laboratory of Biopharmaceutical Production & Formulation Engineering, PLA, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China
Xuexin Duan
State Key Laboratory of Precision Measuring Technology and Instruments, Tianjin University, Tianjin 300072, China
Hongzhi Cao
National Glycoengineering Research Center, State Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, China
Meishan Pei
School of Chemistry and Chemical Engineering, University of Jinan, Jinan 250022, China
Jian-Jun Li
National Key Laboratory of Biochemical Engineering, National Engineering Research Center for Biotechnology (Beijing), Key Laboratory of Biopharmaceutical Production & Formulation Engineering, PLA, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China
Yuguang Du
National Key Laboratory of Biochemical Engineering, National Engineering Research Center for Biotechnology (Beijing), Key Laboratory of Biopharmaceutical Production & Formulation Engineering, PLA, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China
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.
