Preliminary Study On Enzymatic Synthesis Of 6'-sialylactosamine Based On Microfluidic Technology

Glycans of glycoconjugates such as glycoproteins and glycolipids are very important for their proper functions.Glycans research has become the third milestone in exploring the mysteries of life after protein and nucleic acid research.Due to the diversity and complexity of glycans structure,obtaining glycans through purification is still challenging.At present,the best choice for obtaining glycans is still the method of synthesis.Therefore,the development of innovative,environmentally friendly,economical and efficient glycans synthesis technology has important theoretical research on the structure-function relationship of glycans and the development of glycans synthesis instruments.Significance and practical application value.This research will aim to develop an enzyme-catalyzed glycans synthesis technology based on a microfluidic platform,We carried out the following research:(1)Take the enzyme-catalyzed synthesis of N-acetyl-?-2,6-sialyllactosamine as an example,first synthesize avidin-linked substrate(GlcNAc-Biotin ?),and then enzymatically synthesize N-acetyl-?-2,6-Sialyl lactosamine reaction module.Optimize the conditions of the module so that the conversion rate in the first step reaches 90.00%and the conversion rate in the second step reaches 95.00%.The enzyme-catalyzed synthesis reaction using magnetic beads as a solid support supports the solid-phase synthesis of the reaction and optimizes the reaction conditions.Integrate digital microfluidics,solid-phase synthesis and enzyme-catalyzed modular reactions to synthesize Glycans technologies to optimize and improve synthesis methods and conditions.(2)Using GlcNAc-Biotin ? as a substrate to increase the reaction efficiency while adding photosensitive groups to achieve a more efficient and environmentally friendly synthesis process,the optimization of the first step reaction rate increased to 93.70%,the second step reaction rate increased to 97.28%.(3)Achieve ultra-high frequency acoustic microfluidic microfluidic,solid phase purification technology and enzyme catalytic module synthesis of sugar chains and other technical integration,and successfully continuous reaction on the platform to obtain products.The first-step reaction on the chip achieves a conversion rate of 78.00%and the second-step reaction achieves a conversion rate of 91.70%.Continuous reactions can be successfully synthesized and detected.It lays the foundation for the development of an efficient,economical and green automatic Glycans synthesis technology.