Prebiotic properties of Bacillus coagulans MA-13: production of galactoside hydrolyzing enzymes and characterization of the transglycosylation properties of a GH42 β-galactosidase

Abstract

Background The spore-forming lactic acid bacterium Bacillus coagulans MA-13 has been isolated from canned beans manufacturing and successfully employed for the sustainable production of lactic acid from lignocellulosic biomass. Among lactic acid bacteria, B. coagulans strains are generally recognized as safe (GRAS) for human consumption. Low-cost microbial production of industrially valuable products such as lactic acid and various enzymes devoted to the hydrolysis of oligosaccharides and lactose, is of great importance to the food industry. Specifically, α- and β-galactosidases are attractive for their ability to hydrolyze not-digestible galactosides present in the food matrix as well as in the human gastrointestinal tract. Results In this work we have explored the potential of B. coagulans MA-13 as a source of metabolites and enzymes to improve the digestibility and the nutritional value of food. A combination of mass spectrometry analysis with conventional biochemical approaches has been employed to unveil the intra- and extra- cellular glycosyl hydrolase (GH) repertoire of B. coagulans MA-13 under diverse growth conditions. The highest enzymatic activity was detected on β-1,4 and α-1,6-glycosidic linkages and the enzymes responsible for these activities were unambiguously identified as β-galactosidase (GH42) and α-galactosidase (GH36), respectively. Whilst the former has been found only in the cytosol, the latter is localized also extracellularly. The export of this enzyme may occur through a not yet identified secretion mechanism, since a typical signal peptide is missing in the α-galactosidase sequence. A full biochemical characterization of the recombinant β-galactosidase has been carried out and the ability of this enzyme to perform homo- and hetero-condensation reactions to produce galacto-oligosaccharides, has been demonstrated. Conclusions Probiotics which are safe for human use and are capable of producing high levels of both α-galactosidase and β-galactosidase are of great importance to the food industry. In this work we have proven the ability of B. coagulans MA-13 to over-produce these two enzymes thus paving the way for its potential use in treatment of gastrointestinal diseases.

Determination of the Therapeutic Effect of the Use of Bacillus Coagulans in Calf Dyspepsia

“…B. coagulans spores are very hardy, are activated in the stomach, and grow and multiply in the animal's intestine (Xie et al, 2022). In the study by (Aulitto et al, 2021) it was established that B. coagulans can replace lactic acid bacteria in the intestine.…”

A Comparative Analysis of Weizmannia coagulans Genomes Unravels the Genetic Potential for Biotechnological Applications

“…It is worth noting the absence of esxA whose product usually work in association with EssA, EssB, and EssC. Moreover, we have demonstrated in a previous work that WXG-proteins not even related to virulence factors are secreted in MA-13 through a leader-less mechanism, suggesting that esxA is dispensable for this secretion system in this strain [ 15 ]. Interestingly, a homologue (38% identity) of B. subtilis EsxA has been found in some W. coagulans strains (36D1, 2–6 and P38) but there are no reports about its functional characterization in W. coagulans .…”

“…In a recent study, it has been demonstrated that MA-13 over-produces under standard growth conditions α- and β-galactosidases that are key enzymes for improving the nutritional value of lactose-containing food as well as for the production of galactooligosaccharides (GOS) that improve the digestibility and nutritional value of food. Moreover, the identification and comparative analysis of the full repertoire of W. coagulans CAZymes might help to shed light on the probiotic and prebiotic properties of the available strains [ 15 ]. Furthermore, W. coagulans has been also exploited in other diverse biotechnological applications such as in medicine, food, and chemical industry including the production of thermostable enzymes and antimicrobial peptides, such as coagulin [ 16 ].…”

Genomics, Transcriptomics, and Proteomics of SSV1 and Related Fusellovirus: A Minireview

“…For instance, these systems have been successfully used to unveil the role of SSV1 proteins in the infection cycle [ 16 ] and to express heterologous enzymes [ 18 , 75 ]. Finally, a wide variety of biotechnological applications such as medicine, agriculture, pharmacology, cosmetics, electronics, nanotechnology, and environmental safeguarding locate extremophilic microorganisms [ 76 , 77 , 78 , 79 , 80 , 81 , 82 , 83 , 84 , 85 , 86 ] and their viruses as valuable tools [ 2 , 87 , 88 ].…”