2021

Frederiksen, Carsten Østergaard; Cohn, Marianne Thorup; Skov, Lars Kobberøe; Schmidt, Esben Gjerløff Wedebye; Schnorr, Kirk Matthew; Buskov, Steen; Leppänen, Miika; Maasilta, Ilari; Perez-Calvo, Estefania; Lopez-Ulibarri, Rual; Klausen, Mikkel

This study evaluates peptidoglycan hydrolysis by a microbial muramidase from the fungus Acremonium alcalophilum in vitro and in the gastrointestinal tract of broiler chickens. Peptidoglycan used for in vitro studies was derived from 5 gram-positive chicken gut isolate type strains. In vitro peptidoglycan hydrolysis was studied by three approaches: a) helium ion microscopy to identify visual phenotypes of hydrolysis b) reducing end assay to quantify solubilization of peptidoglycan fragments and c) mass spectroscopy to estimate relative abundances of soluble substrates and reaction products. Visual effects of peptidoglycan hydrolysis could be observed by helium ion microscopy and the increase in abundance of soluble peptidoglycan due to hydrolysis was quantified by a reducing end assay. Mass spectroscopy confirmed the release of hydrolysis products and identified muropeptides from the five different peptidoglycan sources. Peptidoglycan hydrolysis in chicken crop, jejunum and caecum samples was measured by quantifying the total and soluble muramic acid content. A significant increase in the proportion of the soluble muramic acid was observed in all three segments upon inclusion of the microbial muramidase in the diet.