Detailed Analysis of Galactooligosaccharides Synthesis with β-Galactosidase from <i>Aspergillus oryzae</i>

Urrutia, Paulina; Rodríguez-Colinas, Bárbara; Fernández‐Arrojo, Lucía; Ballesteros, Antonio; Wilson, Lorena; Illanes, Andrés; Plou, Francisco J.

doi:10.1021/jf304354u

Cited by 120 publications

(79 citation statements)

References 46 publications

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“…The peaks were analyzed using Chromeleon software (Dionex Sunnyvale, CA, USA, 2008)). Identification of the different carbohydrates was done based on commercial standards and previously purified and characterized GOS [19,42,43].…”

Section: High-performance Anion-exchange Chromatography With Pulsed Amentioning

confidence: 99%

Continuous Packed Bed Reactor with Immobilized β-Galactosidase for Production of Galactooligosaccharides (GOS)

et al. 2016

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Abstract:The β-galactosidase from Bacillus circulans was covalently attached to aldehyde-activated (glyoxal) agarose beads and assayed for the continuous production of galactooligosaccharides (GOS) in a packed-bed reactor (PBR). The immobilization was fast (1 h) and the activity of the resulting biocatalyst was 97.4 U/g measured with o-nitrophenyl-β-D-galactopyranoside (ONPG). The biocatalyst showed excellent operational stability in 14 successive 20 min reaction cycles at 45 • C in a batch reactor. A continuous process for GOS synthesis was operated for 213 h at 0.2 mL/min and 45 • C using 100 g/L of lactose as a feed solution. The efficiency of the PBR slightly decreased with time; however, the maximum GOS concentration (24.2 g/L) was obtained after 48 h of operation, which corresponded to 48.6% lactose conversion and thus to maximum transgalactosylation activity.HPAEC-PAD analysis showed that the two major GOS were the trisaccharide Gal-β(1→4)-Gal-β(1→4)-Glc and the tetrasaccharide Gal-β(1→4)-Gal-β(1→4)-Gal-β(1→4)-Glc. The PBR was also assessed in the production of GOS from milk as a feed solution. The stability of the bioreactor was satisfactory during the first 8 h of operation; after that, a decrease in the flow rate was observed, probably due to partial clogging of the column. This work represents a step forward in the continuous production of GOS employing fixed-bed reactors with immobilized β-galactosidases.

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Section: High-performance Anion-exchange Chromatography With Pulsed Amentioning

confidence: 99%

Continuous Packed Bed Reactor with Immobilized β-Galactosidase for Production of Galactooligosaccharides (GOS)

et al. 2016

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“…3-O-β-galactosyl-galactose (3-galactobiose), 4-O-β-galactosyl-galactose (4-galactobiose), 6-O-β-galactosyl-galactose (6-galactobiose), 6-O-β-galactosyl-glucose (allolactose), and 4 -O-β-galactosyl-lactose were acquired from Carbosynth (Berkshire, UK). Other galactooligosaccharide standards were purified according to previous publications [4,22,44]. Other chemicals used for culture medium, buffers, and mobile phases were of analytical grade.…”

Section: Methodsmentioning

confidence: 99%

“…Data acquisition and processing were performed using Chromeleon software. The identification and quantification of different carbohydrates were based on commercially available standards or products purified as described in previous papers [22,35,44].…”

Section: Hpaec-pad Analysismentioning

confidence: 99%

Galactooligosaccharide Production from Pantoea anthophila Strains Isolated from “Tejuino”, a Mexican Traditional Fermented Beverage

Yañez-Ñeco¹,

Rodríguez-Colinas

Amaya-Delgado³

et al. 2017

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Abstract:Two Pantoea anthophila bacterial strains were isolated from "tejuino", a traditional Mexican beverage, and studied as β-galactosidase producers for galactooligosaccharides synthesis. Using 400 g/L of lactose, 50 • C, and 15 U/mL of β-galactosidase activity with ethanol-permeabilized cells, the maximum galactooligosaccharides (GOS) yield determined by High performance anion exchange chromatography with pulse amperometric detection (HPAEC-PAD) was 136 g/L (34% w/w of total sugars) at 96% of lactose conversion for Bac 55.2 and 145 g/L (36% w/w of total sugars) at 94% of lactose conversion for Bac 69.1. The main synthesized products were the disaccharides allolactose [Gal-β(1 → 6)-Glc] and 6-galactobiose [Gal-β(1 → 6)-Gal], as well as the trisaccharides 3 -galactosyl-lactose [Gal-The β-galactosidases present in both strains showed a high transgalactosylation activity and formed principally β(1 → 3) and β(1 → 6) linkages. Considering the stability and bifidogenic properties of GOS containing such types of bonds, P. anthophila strains Bac 55.2 and Bac 69.1 possess a high potential as novel biocatalysts for prebiotic industrial production.

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“…This wavelength was chosen because this enzyme is active at acid pH values [12][13][14], and at these pH conditions, the ε of oNP is negligible at 410 nm. To start the reaction, 50-100 µL of the enzyme solution or suspension were added to 2.5 mL of substrate solution.…”

Section: Standard Determination Of Enzyme Activitymentioning

confidence: 99%

Exploiting the Versatility of Aminated Supports Activated with Glutaraldehyde to Immobilize β-galactosidase from Aspergillus oryzae

et al. 2017

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Abstract:The enzyme β-galactosidase from Aspergillus oryzae has been immobilized in aminated (MANAE)-agarose beads via glutaraldehyde chemistry using different strategies. The immobilization on MANAE-supports was first assayed at different pH values (this gave different stabilities to the immobilized enzymes) and further modified with glutaraldehyde. Dramatic drops in activity were found, even using 0.1% (v/v) glutaraldehyde. The use of a support with lower activation permitted to get a final activity of 30%, but stability was almost identical to that of the just adsorbed enzyme. Next, the immobilization on pre-activated glutaraldehyde beads was assayed at pH 5, 7 and 9. At pH 7, full, rapid immobilization and a high expressed enzyme activity were accomplished. At pH 9, some decrease in enzyme activity was observed. Direct covalent immobilization of the enzyme was very slow; even reducing the volume of enzyme/support ratio, the yield was not complete after 24 h. The stability of the biocatalyst using pre-activated supports was about 4-6 folds more stable than that of the enzyme immobilized via ion exchange at pH 5, with small differences among them. Thus, the immobilization of the enzyme at pH 7 at low ionic strength on pre-activated glutaraldehyde supports seems to be the most adequate in terms of activity, stability and immobilization rate.

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Detailed Analysis of Galactooligosaccharides Synthesis with β-Galactosidase from Aspergillus oryzae

Cited by 120 publications

References 46 publications

Continuous Packed Bed Reactor with Immobilized β-Galactosidase for Production of Galactooligosaccharides (GOS)

Continuous Packed Bed Reactor with Immobilized β-Galactosidase for Production of Galactooligosaccharides (GOS)

Galactooligosaccharide Production from Pantoea anthophila Strains Isolated from “Tejuino”, a Mexican Traditional Fermented Beverage

Exploiting the Versatility of Aminated Supports Activated with Glutaraldehyde to Immobilize β-galactosidase from Aspergillus oryzae

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