Citation
Milano, A. M. T. and Arreola, S. L. B. (2023) Evaluation of transgalactosylation activity of commercial β-galactosidase from Bifidobacterium bifidum for synthesis of prebiotic oligosaccharides. International Food Research Journal (Malaysia), 30. pp. 1450-1460. ISSN 2231 7546
Abstract
Galacto-oligosaccharides (GOS) are products of transgalactosylation reaction of βgalactosidase when lactose is used as the substrate. These carbohydrates are considered prebiotics which stimulate beneficial effects to human health. In the present work, Saphera, a commercial preparation of β-galactosidase from Bifidobacterium bifidum, was biochemically characterised for production of GOS. Using o-nitrophenyl-β-Dgalactopyranoside (oNPG) as the substrate, optimal activity for the enzyme was found to be at pH 6.0 and at 45°C. Ten (10) mM each of either Na+ or K+ enhanced enzyme activity by 10%, while Cu2+, Zn2+, Fe2+, and EDTA showed inhibitory effect on the enzyme activity. When incubated in 50 mM sodium phosphate and pH 6.5, the enzyme was found to have half-life time of 136 ± 6 and 2.1 ± 0.2 h at 30 and 50°C, respectively. The hydrolysis activity of the enzyme predominated when the initial lactose concentration used was 5% (w/v). When initial lactose concentration was increased to 20% (w/v), maximum GOS yield obtained was 10% (w/w) achieved at 86% lactose conversion. Analysis using high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) revealed that the major oligosaccharides produced by the enzyme were 3’-galactosylglucose, 3’-galactosyllactose, 3’-galactobiose, and allolactose, thus showing that this enzyme would prefer to form β-(1→3)- and β-(1→6)-linked GOS. Despite its predominantly hydrolytic activity, Saphera synthesised prebiotic GOS which could be interesting to dairy industry.
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Abstract
Galacto-oligosaccharides (GOS) are products of transgalactosylation reaction of βgalactosidase when lactose is used as the substrate. These carbohydrates are considered prebiotics which stimulate beneficial effects to human health. In the present work, Saphera, a commercial preparation of β-galactosidase from Bifidobacterium bifidum, was biochemically characterised for production of GOS. Using o-nitrophenyl-β-Dgalactopyranoside (oNPG) as the substrate, optimal activity for the enzyme was found to be at pH 6.0 and at 45°C. Ten (10) mM each of either Na+ or K+ enhanced enzyme activity by 10%, while Cu2+, Zn2+, Fe2+, and EDTA showed inhibitory effect on the enzyme activity. When incubated in 50 mM sodium phosphate and pH 6.5, the enzyme was found to have half-life time of 136 ± 6 and 2.1 ± 0.2 h at 30 and 50°C, respectively. The hydrolysis activity of the enzyme predominated when the initial lactose concentration used was 5% (w/v). When initial lactose concentration was increased to 20% (w/v), maximum GOS yield obtained was 10% (w/w) achieved at 86% lactose conversion. Analysis using high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) revealed that the major oligosaccharides produced by the enzyme were 3’-galactosylglucose, 3’-galactosyllactose, 3’-galactobiose, and allolactose, thus showing that this enzyme would prefer to form β-(1→3)- and β-(1→6)-linked GOS. Despite its predominantly hydrolytic activity, Saphera synthesised prebiotic GOS which could be interesting to dairy industry.
Additional Metadata
| Item Type: | Article |
|---|---|
| AGROVOC Term: | Bifidobacterium |
| AGROVOC Term: | enzyme activity |
| AGROVOC Term: | galactosidases |
| AGROVOC Term: | oligosaccharides |
| AGROVOC Term: | prebiotics |
| AGROVOC Term: | experimentation |
| AGROVOC Term: | lactose |
| AGROVOC Term: | dairy industry |
| AGROVOC Term: | human health |
| Geographical Term: | Philippines |
| Uncontrolled Keywords: | beta-galactosidase, galacto-oligosaccharides, lactose, lactase, prebiotics |
| Depositing User: | Nor Hasnita Abdul Samat |
| Date Deposited: | 21 Mar 2025 00:06 |
| Last Modified: | 21 Mar 2025 00:06 |
| URI: | http://webagris.upm.edu.my/id/eprint/1660 |
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