Strategies for enzyme stabilization by intramolecular crosslinking with bifunctional reagents

Fernández‐Lafuente, Roberto; Rosell, Cristina M.; Rodríguez, Verónica; Guisán, José M.

doi:10.1016/0141-0229(94)00090-e

Cited by 151 publications

(72 citation statements)

References 17 publications

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“…That can lead to a conformational changes, and therefore to altered activity of the enzyme if these groups are in vicinity of active site. Also, it has been previously reported that glutaraldehyde had caused the denaturation of immobilized enzyme [13,31,32]. Additionally, it can be seen (Fig.…”

Section: Comparison Of Immobilization Methodsmentioning

confidence: 70%

Immobilization of lipase from Candida rugosa on Sepabeads®: the effect of lipase oxidation by periodates

Prlainović

Knežević‐Jugović

Mijin

et al. 2011

Bioprocess Biosyst Eng

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The objective of this paper was the investigation of a suitable Sepabeads(®) support and method for immobilization of lipase from Candida rugosa. Three different supports were used, two with amino groups, (Sepabeads(®) EC-EA and Sepabeads(®) EC-HA), differing in spacer length (two and six carbons, respectively) and one with epoxy group (Sepabeads(®) EC-EP). Lipase immobilization was carried out by two conventional methods (via epoxy groups and via glutaraldehyde), and with periodate method for modification of lipase. The results of activity assays showed that lipase retained 94.8% or 87.6% of activity after immobilization via epoxy groups or with periodate method, respectively, while glutaraldehyde method was inferior with only 12.7% of retention. The immobilization of lipase, previously modified by periodate oxidation, via amino groups has proven to be more efficient than direct immobilization of lipase via epoxy groups. In such a way immobilized enzyme exhibited higher activity at high reaction temperatures and higher thermal stability.

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Section: Comparison Of Immobilization Methodsmentioning

confidence: 70%

Immobilization of lipase from Candida rugosa on Sepabeads®: the effect of lipase oxidation by periodates

Prlainović

Knežević‐Jugović

Mijin

et al. 2011

Bioprocess Biosyst Eng

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“…33 The primary amino groups of both enzyme and the dendrimer possess on its surface are activated with the molecule of glutaraldehyde. 34 Hence, an intense crosslinking between dendrimer and enzyme seems to be achieved, and this multiple covalent links between the enzyme and dendrimer stabilize the quaternary structure of the protein, and also rigidity of the subunit structures is increased. 35 The intense amino groups on polyamidoamine (PAMAM) surface make the subunit structures of PyOx and GOx to involve in the immobilization.…”

Section: Introductionmentioning

confidence: 99%

Offline glucose biomonitoring in yeast culture by polyamidoamine/ cysteamine‐modified gold electrodes

Yüksel

Akin

Geyik

et al. 2011

Biotechnology Progress

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This article deals with the use of pyranose oxidase (PyOx) and glucose oxidase (GOx) enzymes in amperometric biosensor design and their application in monitoring fermentation processes with the combination of flow injection analysis (FIA). The amperometric studies were carried out at -0.7 V by following the oxygen consumption due to the enzymatic reactions for both batch and FIA modes. Optimization studies (enzyme amounts and pH) and analytical parameters such as linearity, repeatability, effect of interference, storage, and operational stabilities have been studied. Under optimized conditions, for the PyOx-based biosensor, linear graph was obtained from 0.025 to 0.5 mM glucose in phosphate buffer (50 mM) at pH 7.0 with the equation of y = 3.358x + 0.028 and R(2) = 0.998. Linearity was found to be 0.01-1.0 mM in citrate buffer (50 mM and pH 4.0) with the equation of y = 1.539x + 0.181 and R(2) = 0.992 for the GOx biosensor. Finally, these biosensor configurations were further evaluated in a conventional flow injection system. Results from batch experiments provide a guide to design sensitive, stable, and interference-free biosensors for FIA mode. Biosensor stability, dynamic range, and repeatability were also studied in FIA conditions, and the applicability for the determination of glucose in fermentation medium could be successfully demonstrated. The FIA-combined glucose biosensor was used for the offline monitoring of yeast fermentation. The obtained results correlated well with HPLC measurements.

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“…In addition, acid pH might also have a negative impact in enzyme activity. In contrast, in the pre-activated support the activity retention was about 88% (Table 1), likely due to glutaraldehyde crosslinkages as amineglutaraldehyde-amine, amine-glutaraldehydeglutaraldehyde-amine, or amine-(glutaraldehyde) n ,-amine (Fernandez-Lafuente et al 1995). It seemed that glutaraldehyde activation originated dimmers that conferred hydrophobicity to the support surface, and thus improved the retention of lipases with affinity for hydrophobic regions (Barbosa et al 2012).…”

Section: Chemical Attachment Of Lipase Onto Cammentioning

confidence: 99%

Immobilization of Lipases Produced by the Endophytic Fungus Cercospora kikuchii on Chitosan Microparticles

Carneiro

Costa-Silva

Souza

et al. 2014

Braz. arch. biol. technol.

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This work studied the immobilization of Cercospora kikuchii lipases on chitosan microparticles by chemical attachment on chitosan acetate microparticles activated by glutaraldehyde (CAM) added before or after the enzyme and physical adsorption on highly deacetylated chitosan hydrochloride microparticles (CHM). Lipases covalently immobilized on pre-activated CAM showed better performance retaining 88.4% of the enzymatic activity, with 68.2% of immobilization efficiency (IE

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Strategies for enzyme stabilization by intramolecular crosslinking with bifunctional reagents

Cited by 151 publications

References 17 publications

Immobilization of lipase from Candida rugosa on Sepabeads®: the effect of lipase oxidation by periodates

Immobilization of lipase from Candida rugosa on Sepabeads®: the effect of lipase oxidation by periodates

Offline glucose biomonitoring in yeast culture by polyamidoamine/ cysteamine‐modified gold electrodes

Immobilization of Lipases Produced by the Endophytic Fungus Cercospora kikuchii on Chitosan Microparticles

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