Immobilization of Bacillus circulans β-galactosidase and its application in the synthesis of galacto-oligosaccharides under repeated-batch operation

Urrutia, Paulina; Mateo, César; Guisán, José M.; Wilson, Lorena; Illanes, Andrés

doi:10.1016/j.bej.2013.04.015

Cited by 73 publications

(57 citation statements)

References 42 publications

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“…EA obtained with biocatalyst obtained by covalent attachment in glyoxl silica (Glyoxyl-BS) is comparable with those obtained by encapsulation in situ (Table 1) specially with biocatalyst achieved in presence of glucose; showing that this methodology is promissory and competitive. Other reports in the literature for immobilization of β-gal B. circulans different immobilization methodologies, show similar yields to those found in this work [8]. However, the advantage here is related with the immobilization in one step, consuming less energy and generating less waste.…”

Section: Effect Of the Presence Of Sugar On Preparation Of β-Gal/silisupporting

confidence: 85%

“…Synthesis and functionalization of silica was carried out according to reported by Bernal et al [8] using the following molar composition: For the functionalization with glyoxyl groups, 1.0 g silica was contacted with 30 ml of 10% GPTMS aqueous solution, at pH 8.5 and 94°C, during 6 h, under gentle stirring. The hydrolysis of epoxy groups was done with 0.1 M H 2 SO 4 for 2 h at 85°C.…”

Section: β-Gal Immobilization By Covalent Attachment In Glyoxil Silicamentioning

confidence: 99%

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In situ immobilization of β‐galactosidase from Bacillus circulans in silica by sol‐gel process: Application in prebiotic synthesis

Escobar

Illanes

Wilson

et al. 2016

Engineering in Life Sciences

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In situ immobilization of β-galactosidase from Bacillus circulans in silica by sol-gel process: Application in prebiotic synthesisThe enzyme encapsulation is a very well-known stabilization pathway. However, there are some challenges in order to avoid the enzyme denaturation under encapsulation conditions. The β-galactosidase from Bacillus circulans was immobilized through sol-gel encapsulation route assisted by Triton X-100 surfactant and sugars. The effects of sugar presence in the immobilization process and the gelation time on the biocatalyst activity/stability were explained taking into account the characteristics of the formed silica matrix and the changes of the enzyme environment. The enzyme was effectively immobilized by this strategy, with high immobilization yield in terms of activity (29%) and expressed activity (47 IU/g). The immobilization through silica sol-gel in the presence of 1×10 −3 M Triton X-100 and fructose conferred 28.4-fold higher stability to the enzyme compared with the soluble form. This is an advantage for its use in the synthesis of the galacto-oligosaccharides at 50˚C. The total lactose conversion to galacto-oligosaccharides was 26%wt, which is comparable with that reported in the literature. The obtained biocatalyst is useful for the synthesis of galacto-oligosaccharides and its catalytic behavior is rationalized in this work. IntroductionThe β-galactosidase from Bacillus circulans is a hydrolase (E.C.

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Section: Effect Of the Presence Of Sugar On Preparation Of β-Gal/silisupporting

confidence: 85%

Section: β-Gal Immobilization By Covalent Attachment In Glyoxil Silicamentioning

confidence: 99%

In situ immobilization of β‐galactosidase from Bacillus circulans in silica by sol‐gel process: Application in prebiotic synthesis

Escobar

Illanes

Wilson

et al. 2016

Engineering in Life Sciences

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“…As shown, the biocatalyst maintained nearly 100% of its initial activity after 14 cycles (280 min) and the corresponding washings. These results were in accordance with those described by , who analyzed the stability of this enzyme in a batch reactor after immobilization on several mono-and multi-functional supports, including glyoxal agarose [24]. …”

Section: Operational Stability In Batch Reactorsupporting

confidence: 80%

“…This support was previously tested in batch but not in continuous packed-bed reactors (PBR), by Urrutia et al [24]. The operational stability of the immobilized biocatalysts for the production of GOS was assessed.…”

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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“…Furthermore, ␤-galactosidase is used in another large industrial process: the synthesis of galacto-oligosaccharides (GOS), which makes use of the transferase activity of the enzyme [1]. Like many other enzymes, ␤-galactosidase can also be immobilized to extract technical and economic benefits and enable its long time reuse in industrial reactors [2,3].…”

Section: Introductionmentioning

confidence: 99%

Active site titration of immobilized beta-galactosidase for the determination of active enzymes

Jochems

Mueller

Satyawali

et al. 2015

Biochemical Engineering Journal

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Immobilization of Bacillus circulans β-galactosidase and its application in the synthesis of galacto-oligosaccharides under repeated-batch operation

Cited by 73 publications

References 42 publications

In situ immobilization of β‐galactosidase from Bacillus circulans in silica by sol‐gel process: Application in prebiotic synthesis

In situ immobilization of β‐galactosidase from Bacillus circulans in silica by sol‐gel process: Application in prebiotic synthesis

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

Active site titration of immobilized beta-galactosidase for the determination of active enzymes

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