Chia seed mucilage was probed for the presence of structurally diverse free oligosaccharides along with the assessment of hydrolytic and transgalactosylation activities of α-galactosidase from a probiotic bacterium-L...Chia seed mucilage was probed for the presence of structurally diverse free oligosaccharides along with the assessment of hydrolytic and transgalactosylation activities of α-galactosidase from a probiotic bacterium-Lactiplantibacillus plantarum WCFS1(LpαG)on isolated oligosaccharides.Porous graphitic carbon-liquid chromatography-tandem mass spectrometry(PGC LC-MS/MS)analysis revealed the presence of mainly sucrose(disaccharide),planteose(trisaccharide),few isomeric tetrasaccharides as well as lower abundance of isomeric tri-,tetra-,penta-and hexasaccharides.In vitro evaluation of the digestibility of planteose and tetrasaccharides to the intestinal enzymes did not generate monosaccharides.However,LpαG displayed hydrolytic activity towards planteose and selectively acted on two tetrasaccharides,which were identified as sesamose and stachyose.In addition,transglycosylation property of LpαG using planteose led to the synthesis of sesamose.Thus,the hydrolysis of planteose and sesamose was demonstrated for the first time for anα-galactosidase from the probiotic bacterium.Furthermore,the oligosaccharides pool was shown to influence the growth of probiotic L.acidophilus,L.brevis and L.plantarum in vitro,along with the production of short chain fatty acids(SCFAs).These findings would contribute to the potential application of chia seeds to further enhance its health benefits.展开更多
β-Galactosidase was isolated from Ranunculus arvensis seeds using DEAE-cellulose,Sephadex G-100,and Con A sepharose-4B chromatography.The enzyme was purified to electrophoretic homogeneity with a specific activity of...β-Galactosidase was isolated from Ranunculus arvensis seeds using DEAE-cellulose,Sephadex G-100,and Con A sepharose-4B chromatography.The enzyme was purified to electrophoretic homogeneity with a specific activity of 50 U/mg of protein and a yield of 7.1%.The molecular mass of the isolatedβ-galactosidase,as estimated by SDS-PAGE,was 18 kDa,indicating that it was a monomeric form The purifiedβ-galactosidase has a glycoproteinic nature when applied to Con-A-Sepharose-4B chromatography.An activation energy(Ea)of 11 kcal/mol of lactose,pH 5.0,and 50℃ were found to be the optimum parameters to purify β-galactosidase from R.arvensis seeds.The residual activity test was carried at 55-75℃,allowing calculating the half-lives of 533-48 min,enthalpy(ΔH°=110.38-110.21 kJ/mol),free energy(ΔG°=109.88-109.77 kJ/mol),and entropy(ΔS°=1.52-1.26 J/mol·K).The β-galactosidase produced from this species is better than the previously described enzyme due to its kinetic and thermodynamic properties,and it could be used in various industrial applications.Purifiedβ-galactosidase,when incubated with high lactose concentration,showed transgalactosylation activity,leading to trisaccharides as a major product of total galactooligosaccharide(GOS).Therefore,the purifiedβ-galactosidase could be used as a potential alternative in the food industry and would be further explained for trisaccharide synthesis.展开更多
文摘Chia seed mucilage was probed for the presence of structurally diverse free oligosaccharides along with the assessment of hydrolytic and transgalactosylation activities of α-galactosidase from a probiotic bacterium-Lactiplantibacillus plantarum WCFS1(LpαG)on isolated oligosaccharides.Porous graphitic carbon-liquid chromatography-tandem mass spectrometry(PGC LC-MS/MS)analysis revealed the presence of mainly sucrose(disaccharide),planteose(trisaccharide),few isomeric tetrasaccharides as well as lower abundance of isomeric tri-,tetra-,penta-and hexasaccharides.In vitro evaluation of the digestibility of planteose and tetrasaccharides to the intestinal enzymes did not generate monosaccharides.However,LpαG displayed hydrolytic activity towards planteose and selectively acted on two tetrasaccharides,which were identified as sesamose and stachyose.In addition,transglycosylation property of LpαG using planteose led to the synthesis of sesamose.Thus,the hydrolysis of planteose and sesamose was demonstrated for the first time for anα-galactosidase from the probiotic bacterium.Furthermore,the oligosaccharides pool was shown to influence the growth of probiotic L.acidophilus,L.brevis and L.plantarum in vitro,along with the production of short chain fatty acids(SCFAs).These findings would contribute to the potential application of chia seeds to further enhance its health benefits.
基金funding this research work though the project number KKU-IFP2-P-1.
文摘β-Galactosidase was isolated from Ranunculus arvensis seeds using DEAE-cellulose,Sephadex G-100,and Con A sepharose-4B chromatography.The enzyme was purified to electrophoretic homogeneity with a specific activity of 50 U/mg of protein and a yield of 7.1%.The molecular mass of the isolatedβ-galactosidase,as estimated by SDS-PAGE,was 18 kDa,indicating that it was a monomeric form The purifiedβ-galactosidase has a glycoproteinic nature when applied to Con-A-Sepharose-4B chromatography.An activation energy(Ea)of 11 kcal/mol of lactose,pH 5.0,and 50℃ were found to be the optimum parameters to purify β-galactosidase from R.arvensis seeds.The residual activity test was carried at 55-75℃,allowing calculating the half-lives of 533-48 min,enthalpy(ΔH°=110.38-110.21 kJ/mol),free energy(ΔG°=109.88-109.77 kJ/mol),and entropy(ΔS°=1.52-1.26 J/mol·K).The β-galactosidase produced from this species is better than the previously described enzyme due to its kinetic and thermodynamic properties,and it could be used in various industrial applications.Purifiedβ-galactosidase,when incubated with high lactose concentration,showed transgalactosylation activity,leading to trisaccharides as a major product of total galactooligosaccharide(GOS).Therefore,the purifiedβ-galactosidase could be used as a potential alternative in the food industry and would be further explained for trisaccharide synthesis.
