Effect of the type of immobilization of β-galactosidase on the yield and selectivity of synthesis of transgalactosylated oligosaccharides

Guerrero, Cecilia; Aburto, Carla; Súarez, Sebastián; Vera, Carlos; Illanes, Andrés

doi:10.1016/j.bcab.2018.08.021

Cited by 25 publications

(12 citation statements)

References 33 publications

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“…GOS conversion yields obtained with β-gal from A. oryzae using other immobilization methods usually range from 15 to 27% [ 47 ], which highlights the good performance of the biocatalysts herein presented. These are remarkable results since immobilized enzymes usually show poorer performance than the free enzymes due to diffusional limitations [ 48 ]. This supports the notion that nanoflowers, or nanocrystals, are not subjected to significant mass transfer limitations due to their large reactive area, suggesting that the vast majority of the immobilized enzymes are highly exposed to the bulk of the reaction.…”

Section: Resultsmentioning

confidence: 99%

Development of a Hybrid Bioinorganic Nanobiocatalyst: Remarkable Impact of the Immobilization Conditions on Activity and Stability of β-Galactosidase

et al. 2021

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Hybrid bioinorganic biocatalysts have received much attention due to their simple synthesis, high efficiency, and structural features that favor enzyme activity and stability. The present work introduces a biomineralization strategy for the formation of hybrid nanocrystals from β-galactosidase. The effects of the immobilization conditions were studied, identifying the important effect of metal ions and pH on the immobilization yield and the recovered activity. For a deeper understanding of the biomineralization process, an in silico study was carried out to identify the ion binding sites at the different conditions. The selected β-galactosidase nanocrystals showed high specific activity (35,000 IU/g biocatalyst) and remarkable thermal stability with a half-life 11 times higher than the soluble enzyme. The nanobiocatalyst was successfully tested for the synthesis of galacto-oligosaccharides, achieving an outstanding performance, showing no signs of diffusional limitations. Thus, a new, simple, biocompatible and inexpensive nanobiocatalyst was produced with high enzyme recovery (82%), exhibiting high specific activity and high stability, with promising industrial applications.

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Section: Resultsmentioning

confidence: 99%

Development of a Hybrid Bioinorganic Nanobiocatalyst: Remarkable Impact of the Immobilization Conditions on Activity and Stability of β-Galactosidase

et al. 2021

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“…Since both GA and Am-GA are based on the same material, small changes in the diffusing component should be expected and the observed differences between both catalysts might be the result of different intrinsic kinetic parameters. In this regard, Guerrero et al (2018) demonstrated that GA standard, GA fine and Am-GA standard have an apparent Michaelis constant for lactose of sevenfold, threefold and fivefold the value of the Michaelis constant of the free enzyme, respectively. Furthermore, Hoffmann et al (2020) demonstrated that the milieu inside the catalyst particle exerts a significant influence on the selectivity of the reaction, i.e., the ratio between the rate of synthesis and the rate of hydrolysis.…”

Section: <0mentioning

confidence: 95%

Synthesis of Butyl-β-D-Galactoside in the Ternary System: Acetone/1-Butanol/Aqueous Solution

Ahumada

Arenas

Martínez‐Gómez

et al. 2020

Front. Bioeng. Biotechnol.

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The enzymatic synthesis of short-tailed alkyl glucosides is generally carried out in an aqueous-organic biphasic reaction medium with a rather low fatty alcohol concentration in the aqueous phase (where the synthesis occurs). Thus, hydrolytic reactions have a significant impact on the synthesis performance. Given this background, the use of acetone as cosolvent was studied for the synthesis of butyl-β-galactoside with Aspergillus oryzae β-galactosidase. The liquid-liquid equilibrium of the reaction mixture components (acetone/1-butanol/aqueous solution) was determined and the single-and two-phase regions were defined at 30, 40, and 50 • C. It was observed that the liquidliquid equilibrium of the ternary system acetone/1-butanol/water differs significantly from the one obtained using an aqueous solution (50 mM McIlvaine buffer pH 4.5; 5 g L −1) instead of water. This is mainly because of the salting-out effect of the buffer; nevertheless, the presence of lactose also altered the equilibrium. Having this in mind, the effects of temperature (30 and 50 • C) and reaction mixture composition were assessed. Three general conditions were evaluated: single-phase ternary system (30% acetone), two-phase ternary system (10% acetone) and two-phase binary system (0% acetone). Acetone had a deleterious effect on enzyme stability at 50 • C, leading to low reaction yields. However, no enzyme deactivation was detected at 30 • C. Moreover, a reaction yield of 0.98 mol mol −1 was attained in the 30/50/20% (w/w) mixture of acetone/1-butanol/aqueous solution. This very high yield can be explained by the huge increase in the concentration of 1-butanol and the reduction of water activity. The synthesis was carried out using also the β-galactosidase immobilized in glyoxalagarose and amino-glyoxal-agarose, and by aggregation and crosslinking. In the case of agarose-derived catalysts, two average particle diameters were assessed to evaluate the presence of internal mass transfer limitations. Best yield (0.88 mol mol −1) was obtained with glyoxal-agarose derivatives and the particle size had non-effect on yield. The chemical structure of butyl-β-galactoside was determined by NMR and FT-IR.

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“…The combination of a high porous support and the column system allowed a high volumetric activity, more stability against pH changes, and long term continuous operation, while product composition was very similar with FE and IE. Additionally, authors compared the same system design with IE and immobilized whole cells, Glyoxyl-agarose and amino-glyoxyl-agarose derivatives retained the selectivity of the FE for lactulose synthesis while carboxy- Urrutia et al 2013Urrutia et al , 2018Guerrero et al 2017Guerrero et al , 2018 claiming that IE reactor operated more efficiently regarding stability and FOS production. Nishizawa, Nakajima, and Nabetani (2000) tested immobilization of b-fructofuranosidase on ceramic membranes operating in a forced-flow membrane reactor in order to compare physical adsorption and covalent bonding efficiency.…”

Section: Adsorptionmentioning

confidence: 99%

Recent advances in β-galactosidase and fructosyltransferase immobilization technology

Ureta

Martins

Figueira

et al. 2020

Critical Reviews in Food Science and Nutrition

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The highly demanding conditions of industrial processes may lower the stability and affect the activity of enzymes used as biocatalysts. Enzyme immobilization emerged as an approach to promote stabilization and easy removal of enzymes for their reusability. The aim of this review is to go through the principal immobilization strategies addressed to achieve optimal industrial processes with special care on those reported for two types of enzymes: b-galactosidases and fructosyltransferases. The main methods used to immobilize these two enzymes are adsorption, entrapment, covalent coupling and cross-linking or aggregation (no support is used), all of them having pros and cons. Regarding the support, it should be cost-effective, assure the reusability and an easy recovery of the enzyme, increasing its stability and durability. The discussion provided showed that the type of enzyme, its origin, its purity, together with the type of immobilization method and the support will affect the performance during the enzymatic synthesis. Enzymes' immobilization involves interdisciplinary knowledge including enzymology, nanotechnology, molecular dynamics, cellular physiology and process design. The increasing availability of facilities has opened a variety of possibilities to define strategies to optimize the activity and re-usability of b-galactosidases and fructosyltransferases, but there is still great place for innovative developments.

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Effect of the type of immobilization of β-galactosidase on the yield and selectivity of synthesis of transgalactosylated oligosaccharides

Cited by 25 publications

References 33 publications

Development of a Hybrid Bioinorganic Nanobiocatalyst: Remarkable Impact of the Immobilization Conditions on Activity and Stability of β-Galactosidase

Development of a Hybrid Bioinorganic Nanobiocatalyst: Remarkable Impact of the Immobilization Conditions on Activity and Stability of β-Galactosidase

Synthesis of Butyl-β-D-Galactoside in the Ternary System: Acetone/1-Butanol/Aqueous Solution

Recent advances in β-galactosidase and fructosyltransferase immobilization technology

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