Isolation from soil of bacteria producing extracellular alpha galactosidase

Lokuge, M. A.; Mathew, C. Deepal

doi:10.4038/jnsfsr.v28i4.2623

Cited by 3 publications

(3 citation statements)

References 6 publications

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“…However, the electrophoretic mobility of this band (within the 130–180 KDa gel region) was identical to that identified as GH36 (Uniprot code: G2TQE8, Additional file 1 : Table S1) thus indicating that the enzymatic activity revealed by zymography, can be ascribed to the same protein. Besides our experimental evidences, the extracellular localization of the α-galactosidase has been previously described for another closely related B. coagulans strain [ 31 , 45 ] as well as for other soil microorganisms [ 46 ] and for Bacillus megaterium [ 47 ]. It is known that galactomannans are present in seeds of bean and, in general, RFOs (raffinose, stachyose, and verbascose) that contain α 1–6-linked galactose units, are particularly abundant in these legumes [ 9 ].…”

Section: Resultssupporting

confidence: 52%

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

et al. 2021

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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.

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Section: Resultssupporting

confidence: 52%

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

et al. 2021

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“…However, the electrophoretic mobility of this band (within the 130-180 KDa gel region) was identical to that identi ed as GH36 (Uniprot code: G2TQE8, Additional le 1) thus indicating that the enzymatic activity revealed by zymography, can be ascribed to the same protein. Besides our experimental evidences, the extracellular localization of the αgalactosidase has been previously described for another closely related B. coagulans strain [18,32] which was found to be able to grow on galactose-containing polymers (melibiose, ra nose, and stachyose) as well as for other soil microorganisms [33] and for Bacillus megaterium [34]. It is known that galactomannans are present in seeds of beans and in general RFOs (ra nose, stachyose, and verbascose) that contain α 1-6-linked galactose units, are particularly abundant in these legumes [9].…”

Section: Identi Cation Of the Hydrolytic Activities Through Mass Specsupporting

confidence: 74%

Prebiotic Properties of Bacillus Coagulans MA 13: Production of Galactoside Hydrolyzing Enzymes and Characterization of the Transglycosylation Properties of a GH42 β-Galactosidase

Aulitto

Strazzulli

Sansone

et al. 2021

Preprint

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BackgroundThe 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.ResultsIn 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 a 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. ConclusionsProbiotics 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 that are commonly used for treatment of gastrointestinal diseases. Moreover, B. coagulans MA-13 can be employed for an eco-friendly production of prebiotics from dairy food waste because of the ability of β-galactosidase to synthesize galacto-oligosaccharides from lactose.

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“…Extracellular a-galactosidase production of Citrobacter freundii could be increased upto 19mu /ml when cultivated ifi pH. 8 buffer culture medium for ,36h.8…”

Section: Protein Estimationmentioning

confidence: 99%

Kinetic studies of extracellular α galactosidase from Citrobacter freundii

Lokuge

Mathew

2001

J. Natn. Sci. Foundation Sri Lanka

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Extracellular a-galactosidase producing bacteria were isolated from soil. Bacteria that showed a-galactosidase activity were identified as Escherichia coli, Klebsiella pneumonrae and Citrobacter freundii by morphological and biochemical tests. Citrobacter freundii showed the highest enzyme production of 19 milliunitsl ml after 36 h r s of cultivation i n p H 8 phosphate buffer containing peptone. a-Galactosidase from Citrobacter freundii was purified by ammonium sulphatefractionation and DEAE ion exchange chromatography. One a-galactosidase activity peak was observed indicating the presence of a single enzyme form. A 164 fold purification was obtained with a yield of 8%. Polyacrylamide gel electrophoresis of the enzyme showed 2 protein bands. The kinetic properties of the enzyme were studied using p-nitrophenyl a-D-galactopyranoside. The Michaelis constant and maximum reaction velocity obtained were 2.85 x M and 14/mmol/min/mg of protein respectively. Studies on the effect of pH on enzyme activity showed a broad pH optimum from 6.0 to 8.0 with maximum activity a t pH 7.5 a t 2g°C. The enzyme was stable between pH 5.5 to 8.0. The optimum enzyme activity was observed a t 40% a t pH 7.5. The enzyme was stable upto 40 OC. The enzyme preparation did not contain any invertase activity.

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Isolation from soil of bacteria producing extracellular alpha galactosidase

Cited by 3 publications

References 6 publications

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

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

Prebiotic Properties of Bacillus Coagulans MA 13: Production of Galactoside Hydrolyzing Enzymes and Characterization of the Transglycosylation Properties of a GH42 β-Galactosidase

Kinetic studies of extracellular α galactosidase from Citrobacter freundii

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