Carrier-Free Immobilization of α-Galactosidase as Nano-Biocatalysts for Synthesizing Prebiotic α-Galacto-Oligosaccharides
α-Galacto-oligosaccharides (α-GOSs) have great functions as prebiotics and therapeutics. This work established the method of batch synthesis of α-GOSs by immobilized α-galactosidase for the first time, laying a foundation for industrial applications in the future. The α-galactosidase from <i>A...
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doaj-d931e1aa2c28480faa63f837dc1d73f92021-02-26T00:07:57ZengMDPI AGMolecules1420-30492021-02-01261248124810.3390/molecules26051248Carrier-Free Immobilization of α-Galactosidase as Nano-Biocatalysts for Synthesizing Prebiotic α-Galacto-OligosaccharidesYan Liu0Jingyi Yang1Ke Wang2Feiyu Duan3Lili Lu4School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, ChinaSchool of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, ChinaSchool of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, ChinaSchool of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, ChinaSchool of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Chinaα-Galacto-oligosaccharides (α-GOSs) have great functions as prebiotics and therapeutics. This work established the method of batch synthesis of α-GOSs by immobilized α-galactosidase for the first time, laying a foundation for industrial applications in the future. The α-galactosidase from <i>Aspergillus niger</i> L63 was immobilized as cross-linked enzyme aggregates (CLEAs) nano-biocatalyst through enzyme precipitating and cross-linking steps without using carriers. Among the tested agents, the ammonium sulfate showed high precipitation efficacy and induced regular structures of α-galactosidase CLEAs (Aga-CLEAs) that had been analyzed by scanning electron microscopy and Fourier-transform infrared spectroscopy. Through optimization by response surface methodology, the ammonium sulfate-induced Aga-CLEAs achieved a high activity recovery of around 90% at 0.55 U/mL of enzymes and 36.43 mM glutaraldehyde with cross-linking for 1.71 h. Aga-CLEAs showed increased thermal stability and organic solvent tolerance. The storage ability was also improved since it maintained 74.5% activity after storing at 4 °C for three months, significantly higher than that of the free enzyme (21.6%). Moreover, Aga-CLEAs exhibited excellent reusability in the α-GOSs synthesis from galactose, retaining above 66% of enzyme activity after 10 batch reactions, with product yields all above 30%.https://www.mdpi.com/1420-3049/26/5/1248α-galactosidaseimmobilizationcross-linked enzyme aggregatesα-galacto-oligosaccharidesresponse surface methodologybatch synthesis |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yan Liu Jingyi Yang Ke Wang Feiyu Duan Lili Lu |
spellingShingle |
Yan Liu Jingyi Yang Ke Wang Feiyu Duan Lili Lu Carrier-Free Immobilization of α-Galactosidase as Nano-Biocatalysts for Synthesizing Prebiotic α-Galacto-Oligosaccharides Molecules α-galactosidase immobilization cross-linked enzyme aggregates α-galacto-oligosaccharides response surface methodology batch synthesis |
author_facet |
Yan Liu Jingyi Yang Ke Wang Feiyu Duan Lili Lu |
author_sort |
Yan Liu |
title |
Carrier-Free Immobilization of α-Galactosidase as Nano-Biocatalysts for Synthesizing Prebiotic α-Galacto-Oligosaccharides |
title_short |
Carrier-Free Immobilization of α-Galactosidase as Nano-Biocatalysts for Synthesizing Prebiotic α-Galacto-Oligosaccharides |
title_full |
Carrier-Free Immobilization of α-Galactosidase as Nano-Biocatalysts for Synthesizing Prebiotic α-Galacto-Oligosaccharides |
title_fullStr |
Carrier-Free Immobilization of α-Galactosidase as Nano-Biocatalysts for Synthesizing Prebiotic α-Galacto-Oligosaccharides |
title_full_unstemmed |
Carrier-Free Immobilization of α-Galactosidase as Nano-Biocatalysts for Synthesizing Prebiotic α-Galacto-Oligosaccharides |
title_sort |
carrier-free immobilization of α-galactosidase as nano-biocatalysts for synthesizing prebiotic α-galacto-oligosaccharides |
publisher |
MDPI AG |
series |
Molecules |
issn |
1420-3049 |
publishDate |
2021-02-01 |
description |
α-Galacto-oligosaccharides (α-GOSs) have great functions as prebiotics and therapeutics. This work established the method of batch synthesis of α-GOSs by immobilized α-galactosidase for the first time, laying a foundation for industrial applications in the future. The α-galactosidase from <i>Aspergillus niger</i> L63 was immobilized as cross-linked enzyme aggregates (CLEAs) nano-biocatalyst through enzyme precipitating and cross-linking steps without using carriers. Among the tested agents, the ammonium sulfate showed high precipitation efficacy and induced regular structures of α-galactosidase CLEAs (Aga-CLEAs) that had been analyzed by scanning electron microscopy and Fourier-transform infrared spectroscopy. Through optimization by response surface methodology, the ammonium sulfate-induced Aga-CLEAs achieved a high activity recovery of around 90% at 0.55 U/mL of enzymes and 36.43 mM glutaraldehyde with cross-linking for 1.71 h. Aga-CLEAs showed increased thermal stability and organic solvent tolerance. The storage ability was also improved since it maintained 74.5% activity after storing at 4 °C for three months, significantly higher than that of the free enzyme (21.6%). Moreover, Aga-CLEAs exhibited excellent reusability in the α-GOSs synthesis from galactose, retaining above 66% of enzyme activity after 10 batch reactions, with product yields all above 30%. |
topic |
α-galactosidase immobilization cross-linked enzyme aggregates α-galacto-oligosaccharides response surface methodology batch synthesis |
url |
https://www.mdpi.com/1420-3049/26/5/1248 |
work_keys_str_mv |
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