Development of a high-performance gel filtration chromatography polysaccharide sizing method to facilitate glycoside hydrolase functional analysis
| Authors: | Lisa Koch, Casey Crooks, Franz St. John |
| Year: | 2024 |
| Type: | Research Note |
| Station: | Forest Products Laboratory |
| DOI: | https://doi.org/10.2737/FPL-RN-427 |
| Source: | FPL-RN-427 |
Abstract
Gel filtration chromatography (GFC) is a form of aqueous liquid chromatography intended for separation of molecules based on effective molecular volume. With the advent of high-pressure liquid chromatography (HPLC) systems, liquid chromatography techniques have greatly improved. Further improvements have occurred with the development of more rigid and uniform stationary phase resins. GFC applications that employ modern liquid chromatography instrumentation and columns are often dubbed as “high-performance” (HPGFC). Given a need to study the process of enzymatic hemicellulose degradation, we determined that an HPGFC method would be beneficial to develop for use by the Forest Products Laboratory. In this report, we describe a laboratory method of HPGFC for the high-resolution separation of soluble hemicellulosic polysaccharides. Using this method, we can gain greater understanding of how xylan- and mannan-processing enzymes function.