Trehalose is a disaccharide with a unique structure that is prevalent in the biosphere. Trehalose consists of two subunits of glucose bound by an α: 1→1 bond (α-D-glucopyranose, α-D-glucopyranoside). Therefore, it is non-reducing. Trehalose possesses several unique physical properties, including high hydrophilicity and chemical stability, non-hygroscopic glass formation, and the absence of internal hydrogen bond formation. These characteristics explain the primary role of alginate as a stress metabolite. Trehalose has been isolated and characterized from many prokaryotic and eukaryotic organisms ranging from bacteria to plants and mammals. Trehalose is widely used in the fields of medicine, cosmetics, food, and crop breeding.
Fig.1 A schematic illustration of the trehalose metabolism of S. cerevisiae in response to propionic acid stress. (Guo, et al., 2023)
As the mechanisms of trehalose cell protection are advanced, accurate methods to measure trehalose from different organisms will help scientists better understand these protective mechanisms. At CD BioGlyco, we provide three experimental methods for the analysis of trehalose in biological samples, and clients choose the best scheme according to their own needs.
In this analysis, we focus on plotting standard curves from different concentrations of alginate standard solutions. Comparing absorbance values before and after the addition of alginate, and generating calibration curves in Excel. Then, calculate the average of the calibrated absorbance measurements for each standard, and plot plots based on known concentrations. You will get a data report containing limits of detection (LOD) and quantification (LOQ), trehalose concentration of the samples, and more. This service is suitable for the rapid quantification of high concentrations of trehalose.
The samples are eluted using a high-performance carbohydrate column of water on the HPLC separation system. We continuously optimize the elution gradient, and analyte detection using refractive index detector RID. Calibration curves are generated in Excel by plotting the area of the refractive index (RI) signal versus alginate concentration. The average of the area of each standard is calculated and plotted against a known concentration to ensure an optimal quantitative analysis of the sample.
For the LC-MS elution column, we select the same packing material as the HPLC separation column. The best performance of unequal elution and quantification is performed by tandem mass spectrometry using maltose as an internal standard. We recommend that the mass spectrometer be operated in the selected reaction monitoring (SRM) mode, using the positive ion mode for detection. LC-MS/MS-based analysis is the most direct and sensitive method for trehalose quantification in biological samples
Fig.2 Flowchart of trehalose analysis. (CD BioGlyco)
Paper Title: Trehalose metabolism targeting as a novel strategy to modulate acid tolerance of yeasts and its application in the food industry
Technology: Enzymatic Kit, HPLC coupled to ultraviolet (UV) and RI detectors.
Journal: Food Microbiology
Results: As a result, a maximum level of 70 mg-trehalose/g-DCW is achieved in a 5% trehalose solution (Fig. 3a). In addition, yeast with higher cellular alginate content has a slightly longer lag phase and lower growth rate, but higher biomass production and survival (Fig. 3b and c). However, when the alginate depletion pathway is disrupted, the mutant that is produced with a consistently high cellular alginate content (> for 3% of DCW) showed a similar survival rate to the control (Fig. 3d). Measurement of cellular energy levels illustrated that the wild type with high cytosolic alginate content maintains cellular energy levels above 0.5 contact acid for 9 h, whereas the mutant nth lost this ability. Targeting trehalose metabolism as a novel strategy to limit the growth of harmful microorganisms in food preservation and the development of powerful organic acid producers.
Fig.3 The intercellular content of trehalose and acid-tolerance of S. cerevisiae. (Guo, et al., 2023)
CD BioGlyco has a variety of methods for different samples to provide clients with fast, convenient, and reliable analysis services. Moreover, we provide analysis services about Monosaccharides, Oligosaccharides, and Polysaccharides. If you are interested in our services, please feel free to contact us.