The Quantitative Measurement of Peptidoglycan Components Obtained from Acidic Hydrolysis in Gram-Positive and Gram-Negative Bacteria via Hydrophilic Interaction Liquid Chromatography Coupled with Mass Spectrometry

The high throughput in genome sequencing and metabolic model (MM) reconstruction has democratised bioinformatics approaches such as flux balance analysis. Fluxes’ prediction accuracy greatly relates to the deepness of the MM curation for a specific organism starting from the cell composition. One component is the cell wall, which is a functional barrier (cell shape, exchanges) with the environment. The bacterial cell wall (BCW), including its thickness, structure, and composition, has been extensively studied in Escherichia coli but poorly described for other organisms. The peptidoglycan (PG) layer composing the BCW is usually thinner in Gram− bacteria than in Gram+ bacteria. In both bacteria groups, PG is a polymeric mesh-like structure of amino acids and sugars, including N-acetylglucosamine, N-acetylmuramic acid, and amino acids. In this study, we propose a high-throughput method to characterise and quantify PG in Gram-positive and Gram-negative bacteria using acidic hydrolysis and hydrophilic interaction liquid chromatography coupled with mass spectrometry (HILIC-MS). The method showed a relatively short time frame (11 min analytical run), low inter- and intraday variability (3.2% and 4%, respectively), and high sensitivity and selectivity (limits of quantification in the sub mg/L range). The method was successfully applied on two Gram-negative bacteria (Escherichia coli K12 MG1655, Bacteroides thetaiotaomicron DSM 2079) and one Gram-positive bacterium (Streptococcus salivarius ssp. thermophilus DSM20259). The PG concentration ranged from 1.6% w/w to 14% w/w of the dry cell weight. The results were in good correlation with previously published results. With further development, the PG concentration provided by this newly developed method could reinforce the curation of MM

Microbiological, chemical, and sensorial characterisation of commercially available plant-based yoghurt alternatives

Consumer demand for plant-based dairy alternatives has increased rapidly during the past few years and the market has been saturated with a wide variety of alternative products. The general aim of this study was to broaden the understanding of the composition and characteristics of currently commercially available plant-based yoghurt alternatives focusing especially on the content of live bacteria. The bacterial composition, including the content of live bacteria in yoghurt alternatives, was evaluated using metagenetic sequencing of the 16S rRNA gene amplicons in combination with the novel PMAxx treatment approach. The content of organic acids, sugars, and volatiles was measured, and descriptive sensory analysis was carried out to comprehensively describe the products. While the main ingredient (soya, oat, coconut, or lupin) determined the general characteristics of the product, significant differences were observed in both chemical and microbiological composition and sensorial attributes even among the yoghurt alternatives made from the same plant ingredient

Starter culture growth dynamics and sensory properties of fermented oat drink

In the present study, an oat drink, a plant-based alternative to dairy products, was developed by fermenting the oat base with different vegan starter cultures. The desired pH below 4.2 was achieved in 12 h, regardless of starter culture used. Metagenomic sequencing revealed that S. thermophilus was the dominating species, ranging from 38% to 99% of the total microbial consortia. At lower pH values, population of L. acidophilus, L. plantarum and L. paracasei continued to increase in fermented oat drinks. Lactic acid was produced between 1.6 and 2.8 g/L. The sensory panel showed that all fermented oat drinks had a sour odor and taste. The volatile compounds identified belonged to the ketone, alcohol, aldehyde, acids, and furan classes. The concentration of the most preferred volatile components, such as diacetyl and acetoin, increased during fermentation. However, sensory evaluation showed that all samples were associated with cereals and not dairy in terms of taste and odor. Rheological analysis showed the formation of weak gel-like structures in fermented oat drinks. Overall, fermentation improved flavor and texture of the product. This study provides a broad overview of the oat drink fermentation process from the perspectives of starter culture growth, microbial consortium dynamics, lactic acid bacteria metabolism, and sensory profile formation