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Thermostable β-galactosidases for the synthesis of human milk oligosaccharides.
- Source :
-
New Biotechnology . May2016, Vol. 33 Issue 3, p355-360. 6p. - Publication Year :
- 2016
-
Abstract
- Human milk oligosaccharides (HMOs) designate a unique family of bioactive lactose-based molecules present in human breast milk. Using lactose as a cheap donor, some β-galactosidases (EC 3.2.1.23) can catalyze transgalactosylation to form the human milk oligosaccharide lacto- N -neotetraose (LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc). In order to reduce reaction times and be able to work at temperatures, which are less welcoming to microbial growth, the current study investigates the possibility of using thermostable β-galactosidases for synthesis of LNnT and N -acetyllactosamine (LacNAc; Gal-β(1,4)-GlcNAc), the latter being a core structure in HMOs. Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus , were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. The three β-galactosidases were compared in the current study in terms of their transgalactosylation activity in the formation of LacNAc and LNnT. In all cases, BgaD-D was the most potent transgalactosidase, but both thermostable GH 1 β-galactosidases could catalyze formation of LNnT and LacNAc, with Ttβ-gly giving higher yields than CelB. The thermal stability of the three β-galactosidases was elucidated and the results were used to optimize the reaction efficiency in the formation of LacNAc, resulting in 5–6 times higher reaction yields and significantly shorter reaction times. [ABSTRACT FROM AUTHOR]
- Subjects :
- *GALACTOSIDASES
*THERMOSTAT
*BREAST milk
*OLIGOSACCHARIDES
*THERMUS thermophilus
Subjects
Details
- Language :
- English
- ISSN :
- 18716784
- Volume :
- 33
- Issue :
- 3
- Database :
- Academic Search Index
- Journal :
- New Biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 113826414
- Full Text :
- https://doi.org/10.1016/j.nbt.2016.01.003