High throughput exopolysaccharide screening platform: From strain cultivation to monosaccharide composition and carbohydrate fingerprinting in one day

Abstract

Microbial exopolysaccharides (EPS) are multifunctional biogenic polymers, which exist in highly diverse chemical structures. To facilitate a fast determination of the carbohydrate composition of novel isolated strains or modified EPS variants a fast screening and analytical method is required. The platform as realized and described in this article is based on the fast carbohydrate analysis via liquid chromatography coupled with ultra violet and electrospray ionization ion trap detection in 96-well format to detect different sugars, sugar derivatives and substituents such as pyruvate. Monosaccharide analysis from hydrolyzed polysaccharides was validated successfully by 16 commercially available polymers with known structure. The method is sensitive enough to distinguish various types of sphingans which solely differ in small alterations in the monomer composition. Even a quantitative detection of single monomers as present in complex plant polysaccharides like karaya gum, with the lowest recovery, was in accordance with literature. Furthermore, 94 bacterial strains for the validation of the screening platform were completely analyzed and 41 EPS producing strains were efficiently identified. Using the method a carbohydrate-fingerprint of the strains was obtained even allowing a very fast differentiation between strains belonging to the same species. This method can become a valuable tool not only in the fast analysis of strain isolates but also in the targeted screening for polysaccharides containing special rare sugars as well in the screening of strain libraries from genetic engineering for altered structures.

Bacterial exopolysaccharides: biosynthesis pathways and engineering strategies

“…This phenomenon of the genetic equipment for the production of more than one EPS is also very widespread amongst microbes ( Christensen et al, 1985 ; Sutherland, 2001 ; Wozniak et al, 2003b ; Laue et al, 2008 ) and complicates the defined analysis of sugar moieties making up the polymer ( Rühmann et al, 2015 ).…”

Automated Modular High Throughput Exopolysaccharide Screening Platform Coupled with Highly Sensitive Carbohydrate Fingerprint Analysis

“…Furthermore, the validation of the gel-filtration and the carbohydrate fingerprint for various genera have been described in another publication 6 . In sum, the screening platform with its modular structure can easily be modified and adapted to individual requirements of the user.…”

Enzymatic Transformations Involved in the Biosynthesis of Microbial Exo‐polysaccharides Based on the Assembly of Repeat Units

“…This method would have the potential to expand the portfolioo fs tructurally divers EPS variants. [71] By using the power of next-generation sequencing technologies [72] in combination with highly exact and fast analytical techniques [73,74] as well as synthetic biological methods, the potentialt ot ailor make EPSs might be fully exploited, and new fields of applications could be opened.…”