Optimization by experimental design of polyacrylamide gel composition as support for enzyme immobilization by entrapment

Pizarro, Consuelo; Fernández-Torroba, M. A.; Benito, Carmen; González-Sáiz, J.M.

doi:10.1002/(sici)1097-0290(19970305)53:5<497::aid-bit7>3.0.co;2-c

Cited by 39 publications

(12 citation statements)

References 17 publications

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“…This section makes a comparison between the immobilisation by covalent linking presented in this study and immobilisation by physical entrapment studied by Pizarro et al 16 using the same methodology, the same support, enzyme, initiator and other variables proposed here.…”

Section: Resultsmentioning

confidence: 95%

“…Behaviour of the gel in the solution : the swelling ratio and porosity were equal to the size and the shape of the particles of gel with the immobilised enzymes studied by Pizarro et al 16, 21 In this study, gels in a wafer with a diameter of 10 mm have been used.…”

Section: Resultsmentioning

confidence: 99%

“…This permits the composition of the gel to be developed in terms of enzyme and substrate size, also taking into account the surroundings or reaction media. An optimised gel would allow immobilisation without the risk of the enzymes leaving the polymer matrix, but without producing serious diffusional resistances 16…”

Section: Introductionmentioning

confidence: 99%

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Optimisation of polyacrylamide gel composition as support for enzyme immobilisation by covalent linking

Pizarro

González-Sáiz

Sanchez-Jimenez

2000

J. Chem. Technol. Biotechnol.

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A methodology based on experimental design to optimise a polyacrylamide gel as the support for enzyme immobilisation by covalent linking is described. A proper selection of the variables responsible for the gel structure, the amount of the linking agent, the support enzyme and suitable synthesis conditions lead to an increase in the activity retained by the gel. The maximum retained activity is chosen with the response surface methodology in terms of gel composition.

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Section: Resultsmentioning

confidence: 95%

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Optimisation of polyacrylamide gel composition as support for enzyme immobilisation by covalent linking

Pizarro

González-Sáiz

Sanchez-Jimenez

2000

J. Chem. Technol. Biotechnol.

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“…Thus, the enzyme is subjected to a strong electrostatic field which may affect its mode of action . PAAm‐derived gels are cross‐linked networks of synthetic polymers with high molecular weight .…”

Section: Carrier Materialsmentioning

confidence: 99%

Immobilization of Lipases – A Review. Part II: Carrier Materials

Thangaraj

Solomon

2019

ChemBioEng Reviews

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Carriers used in the immobilization process play a major role in enhancing the properties of the biocatalyst. Polymers are used as organic carriers for enzyme immobilization to improve the thermal, chemical, and operational stability of the enzyme. A variety of carriers are used in the immobilization of enzymes, e.g., mica, silica, zeolites, hydrotalcites, activated carbon, gold and magnetic nanoparticles. Silica‐based carriers offer suitable matrices for enzyme immobilization to manufacture industrial products. Immobilization on nanoparticles has become attractive in biocatalytic applications due to the combination of physical, chemical, catalytic, electronic, and optical properties. Gold nanoparticles also have received attention in the preparation of biocatalysts due to the above‐mentioned reasons. Magnetic nanoparticles are used as a carrier since they can be isolated by an external magnetic field, which is of special interest in the synthesis of organic products such as biodiesel. Different carrier materials involved in the immobilization of enzymes are discussed in this paper.

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“…Formation of PAA gels involves the cross-linking of acrylamide and bis-acrylamide monomers in a radical polymerization process employing ammonium peroxodisulfate (APS) and N,N,N¢,N¢-tetramethylethylenediamine (TEMED) as radical-forming agent and enhancer, respectively [19,20]. The acrylamide concentration for the entrapment of enzymes can range between 5% (w ⁄ v) [10] and 30% (w ⁄ v) [21], depending on the required network density [4].…”

Section: Effects Of Paa Building Blocks On Wild-type Fdhmentioning

confidence: 99%

Directed evolution of formate dehydrogenase from Candida boidinii for improved stability during entrapment in polyacrylamide

et al. 2006

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In two cycles of an error‐prone PCR process, variants of formate dehydrogenase from Candida boidinii were created which revealed an up to 4.4‐fold (440%) higher residual activity after entrapment in polyacrylamide gels than the wild‐type enzyme. These were identified in an assay using single precursor molecules of polyacrylamide instead of the complete gel for selection. The stabilization resulted from an exchange of distinct lysine, glutamic acid, and cysteine residues remote from the active site, which did not affect the kinetics of the catalyzed reaction. Thermal stability increased at the exchange of lysine and glutamic acid, but decreased due the exchange of cysteine. Overall, the variants reveal very suitable properties for application in a technical synthetic process, enabling use of entrapment in polyacrylamide as an economic and versatile immobilization method.

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Optimization by experimental design of polyacrylamide gel composition as support for enzyme immobilization by entrapment

Cited by 39 publications

References 17 publications

Optimisation of polyacrylamide gel composition as support for enzyme immobilisation by covalent linking

Optimisation of polyacrylamide gel composition as support for enzyme immobilisation by covalent linking

Immobilization of Lipases – A Review. Part II: Carrier Materials

Directed evolution of formate dehydrogenase from Candida boidinii for improved stability during entrapment in polyacrylamide

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