Martina Aulitto1,2, Andrea Strazzulli1,3, Ferdinando Sansone1, Flora Cozzolino4,5, Maria Monti4,5, Marco Moracci1,3,6, Gabriella Fiorentino1,7, Danila Limauro1,7, Simonetta Bartolucci1, Patrizia Contursi8,9,10. 1. Department of Biology, University of Naples Federico II, 80126, Naples, Italy. 2. Division of Biological Systems and Engineering, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA. 3. Task Force On Microbiome Studies, University of Naples Federico II, Naples, Italy. 4. Department of Chemical Sciences, University of Naples Federico II, 80126, Naples, Italy. 5. CEINGE Advanced Biotechnologies, University of Naples Federico II, 80145, Naples, Italy. 6. Institute of Biosciences and BioResources-National Research Council of Italy, Naples, Italy. 7. BAT Center-Interuniversity Center for Studies On Bioinspired Agro-Environmental Technology, University of Napoli Federico II, Portici, NA, Italy. 8. Department of Biology, University of Naples Federico II, 80126, Naples, Italy. contursi@unina.it. 9. Task Force On Microbiome Studies, University of Naples Federico II, Naples, Italy. contursi@unina.it. 10. BAT Center-Interuniversity Center for Studies On Bioinspired Agro-Environmental Technology, University of Napoli Federico II, Portici, NA, Italy. contursi@unina.it.
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.
BACKGROUND: The spore-forming lactic acid bacterium Bacillus coagulansMA-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. coagulansMA-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. coagulansMA-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. coagulansMA-13 to over-produce these two enzymes thus paving the way for its potential use in treatment of gastrointestinal diseases.
Authors: Martina Aulitto; Salvatore Fusco; David Benjamin Nickel; Simonetta Bartolucci; Patrizia Contursi; Carl Johan Franzén Journal: Biotechnol Biofuels Date: 2019-02-28 Impact factor: 6.040
Authors: Giovanni Gallo; Rosanna Puopolo; Miriam Carbonaro; Emanuela Maresca; Gabriella Fiorentino Journal: Int J Environ Res Public Health Date: 2021-05-14 Impact factor: 3.390