High-resolution method for isocratic HPLC analysis of inulin-type fructooligosaccharides

Area of Science:

• Microbiology
• Biotechnology
• Analytical Chemistry

Background:

• Gut microbiota modulation is key for human health, with prebiotics playing a significant role.
• Oligosaccharides are dominant prebiotics, selectively promoting beneficial gut bacteria.
• Microbial enzymes, like inulosucrases from Lactobacillus, are explored for prebiotic synthesis.

Purpose of the Study:

• To efficiently synthesize inulin-type fructooligosaccharides (inulin-FOS) using a recombinant enzyme.
• To develop a rapid and resource-efficient HPLC method for inulin-FOS analysis.

Main Methods:

• Utilized a truncated inulosucrase (InuGB-V3) from Lactobacillus gasseri DSM 20604 for inulin-FOS synthesis.
• Optimized an isocratic HPLC method for analyzing inulin-FOS with varying degrees of polymerization (DP).

Main Results:

• Achieved high product titers of 146.2 ± 7.4 g/L inulin-FOS.
• Developed an HPLC method for rapid analysis (under 6 minutes for DP up to 5) and high-resolution separation of long-chain inulin-FOS (DP 17 in under 45 minutes).

Conclusions:

• The recombinant inulosucrase enables efficient inulin-FOS production.
• The developed isocratic HPLC method is sensitive, cost-effective, and provides high-resolution separation for prebiotic analysis.