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A Simple Enzymatic Method for Quantitation of 2'-Fucosyllactose
Emine Seydametova1, Jonghyeok Shin1, Jiwon Yu1
1Department of Integrative Biotechnology, College of Biotechnology and Bioengineering, Sungkyunkwan University, Suwon 16419, Republic Korea.
Journal of Microbiology and Biotechnology
|June 22, 2018
Summary
A new microplate assay simplifies quantifying 2'-Fucosyllactose (2'-FL), a key human milk oligosaccharide. This efficient method allows simultaneous analysis of numerous samples, aiding infant health research.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Nutritional Science
Background:
- 2 -Fucosyllactose (2 -FL) is a crucial human milk oligosaccharide with significant infant health benefits.
- Quantifying 2 -FL is important for nutritional and health studies.
- Existing methods like HPLC are not suitable for high-throughput sample analysis.
Purpose of the Study:
- To develop a simple, high-throughput method for quantifying 2 -FL.
- To enable simultaneous detection of 2 -FL in a large number of samples.
Main Methods:
- A two-step enzymatic assay was developed for 2 -FL quantification.
- Step 1: Enzymatic release of L-fucose from 2 -FL using α-(1-2,3,4,6)-L-fucosidase.
- Step 2: Measurement of NADPH produced during L-fucose oxidation by L-fucose dehydrogenase.
Main Results:
- The method quantifies 2 -FL in a 96-well microplate format.
- Quantification is achieved within 50 minutes for concentrations up to 5 g/l.
- The assay is efficient and simple, suitable for analyzing numerous samples concurrently.
Conclusions:
- The developed microplate assay provides a simple and efficient method for 2 -FL quantification.
- This high-throughput method facilitates the analysis of large sample sets.
- The assay is valuable for research on human milk oligosaccharides and infant nutrition.