Immobilization of thermostable β-galactosidase on epoxy support and its use for lactose hydrolysis and galactooligosaccharides biosynthesis

Marín-Navarro, Julia; Talens-Perales, David; Oude-Vrielink, Anneloes; Cañada, F. Javier; Polaina, Julio

doi:10.1007/s11274-013-1517-8

Cited by 37 publications

(42 citation statements)

References 49 publications

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“…Covalent immobilization has the advantage of forming robust linkages between the enzyme and the carrier, which minimizes the loss of activity caused by enzyme leakage from the support [10], extends its lifetime by protecting the three-dimensional structure of the protein [11] and allows the development of continuous processes. The use of immobilized β-galactosidases in continuous bioreactors has mostly been applied to the hydrolysis of lactose in cheese whey [12][13][14].…”

Section: Immobilization Of β-Galactosidase From B Circulans On Glyoxmentioning

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: Immobilization Of β-Galactosidase From B Circulans On Glyoxmentioning

confidence: 99%

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

et al. 2016

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“…Therefore, new technologies have been developed for the production of milk products with low lactose content in order to satisfy the need for people suffering from lactose intolerance (Relpius, 2008). Different methods have recently been proposed in order to remove lactose from milk, as such lactose hydrolysis, capillary electrophoresis, ultrafiltration, and ion-exchange chromatography (Navarro et al, 2014;Bargeman, 2003;Marín-Navarro et al, 2014;Harju, 2007;Mohammad et al, 2012;Oliveira et al, 2014). Other studies have shown the feasibility of using the molecular recognition ability of molecularly imprinted polymers (MIPs) in processes such as adsorption using polymer or silica-based imprinted materials for proteins, polysaccharides and nucleic acids as templates (Li et al, 2014;Escobar and Santos, 2014;Dickert and Lieberzeit, 2006;Turner et al, 2006;Bossi et al, 2007;Bergmann and Peppas 2008;Kryscio and Peppas, 2012;Whitcombe et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

Adsorption Process of Molecularly Imprinted Silica for Extraction of Lactose From Milk

Balieiro

Santos

Pereira

et al. 2016

Braz. J. Chem. Eng.

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-In Brazil, about 25-30% of the population has some degree of intolerance to lactose, a disorder associated with the inability of the body to digest lactose due to a disability or absence of the enzyme lactase. The goal of this study was to evaluate the performance of adsorption of lactose from fresh milk using a fixed bed column of molecularly imprinted polymer (MIP). The polymeric material was characterized using Scanning electron microscopy (SEM) analysis, thermal analysis (e.g., differential scanning calorimetric (DSC) and thermogravimetric analysis (TGA), Fourier Transform Infrared Spectroscopy (FTIR), and the method of Braunauer, Emmet and Teller (BET). The adsorption column dynamics and performance were studied by the breakthrough curves using a 2 4-1 fractional factorial design. The chemical and structural characterization of the pure matrix and imprinted polymers confirmed the molecularly imprinted polymer (MIP) imprinted with lactose. The highest capacity was 62.21 mgg -1 , obtained at 307.1 K and a flow rate of 12.5 mL.min -1 , with central point conditions, 320.1 K and 9 mL.min -1 , with an average value of 50.9 mg.g -1 . The results indicate that the molecularly imprinted polymer is efficient.

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“…Comparative docking analysis with a DA type 3 enzyme (TmLac) and a DA type 5 enzyme (BgaD-D) may add some hints about the structural basis for this different product specificity. Previous studies with TmLac showed that the main transglycosylation product of this enzyme is β-D-(1,3)-galactosyl-lactose, with a minor production of β-D-(1,6)-galactosyl-lactose [47]. Site-directed mutagenesis and structural modelling of TmLac suggested that Trp 959 (Fig 5B) functions as the aromatic binding platform for the acceptor lactose in the synthesis of the β-(1,3) GOS [4].…”

Section: Discussionmentioning

confidence: 99%

Analysis of Domain Architecture and Phylogenetics of Family 2 Glycoside Hydrolases (GH2)

et al. 2016

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In this work we report a detailed analysis of the topology and phylogenetics of family 2 glycoside hydrolases (GH2). We distinguish five topologies or domain architectures based on the presence and distribution of protein domains defined in Pfam and Interpro databases. All of them share a central TIM barrel (catalytic module) with two β-sandwich domains (non-catalytic) at the N-terminal end, but differ in the occurrence and nature of additional non-catalytic modules at the C-terminal region. Phylogenetic analysis was based on the sequence of the Pfam Glyco_hydro_2_C catalytic module present in most GH2 proteins. Our results led us to propose a model in which evolutionary diversity of GH2 enzymes is driven by the addition of different non-catalytic domains at the C-terminal region. This model accounts for the divergence of β-galactosidases from β-glucuronidases, the diversification of β-galactosidases with different transglycosylation specificities, and the emergence of bicistronic β-galactosidases. This study also allows the identification of groups of functionally uncharacterized protein sequences with potential biotechnological interest.

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Immobilization of thermostable β-galactosidase on epoxy support and its use for lactose hydrolysis and galactooligosaccharides biosynthesis

Cited by 37 publications

References 49 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)

Adsorption Process of Molecularly Imprinted Silica for Extraction of Lactose From Milk

Analysis of Domain Architecture and Phylogenetics of Family 2 Glycoside Hydrolases (GH2)

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