Enhanced Bioactivity of Bifidobacterium spp. in the Presence Of Milk Fat Globule Membrane
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Date
2022-05
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The Ohio State University
Abstract
Bifidobacterium is a diverse genus of bacteria with a high potential for creating powerful probiotics. Strains of Bifidobacterium can be isolated from mammalian intestines as a part of the natural microbiome, making them naturally coexistent with human biology, and making them a natural choice for enhancement. Additionally, milk fat globule membrane (MFGM), a component of milk containing numerous nutrients necessary for every living being is speculated to be capable of increasing the bioactivity of Bifidobacterium both through its own antibacterial properties and through improving the effectiveness of Bifidobacterium spp. production of antibacterial compounds. This study sought to measure the bioactivity of several Bifidobacterium spp. and analyze any improvements that could be noted after the introduction of MFGM. Sample strains were initially grown in CGB (commercially accepted broth used for specific growth of Bifidobacterium) broth before being moved to larger volumes of either CGB broth or MFGM infused CGB broth. Samples grown in MFGM infused broth showed significantly more growth over a 24-hour incubation period, with this being the first hint at increased activity in the presence of MFGM. Initial samples from CGB broth were used in SDS PAGE and zymography assays to assess the size of active enzymes in the strains, this showed that Bifidobacterium longum subsp. infantis had several active enzymes in the 40-150 kDa range while Bifidobacterium animalis subsp. lactis seemed to at least have one band of activity in the 100 kDa range. Enzymatic assays revealed strains had varying, but high levels of PGH (peptidoglycan hydrolases) activity in the cells of each strain—particularly with endopeptidase in B. longum, amidase in B. bifidum, and glucoamidase in B. longum subsp. infantis. In an agar diffusion test, B. longum subsp. infantis showed a high capability for bacterial inhibition against Listeria innocua, both in cellular samples and in supernatant protein samples. Furthermore, Bradford assay revealed that supernatant protein samples grown in MFGM broth could contain between 10 and 32 times more protein when compared to samples grown in CGB broth. Exopolysaccharides may be a component of biofilms produced by Bifidobacterium spp. which can only be created for some strains in the presence of MFGM, this is notable as many strains produce a pink-stained biofilm after Gram Staining. MFGM has a clear effect on the bioactivity of Bifidobacterium spp., further research is needed to gain more detailed insight into the specifics of the activities explored in this research.
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Keywords
Bifidobacterium, Bioactivity, Enhanced, MFGM, Milk Fat Globule Membrane