ChemInform Abstract: Immobilized Enzymes and Cells

Kennedy, J. F.; Melo, E. H. M.; Jumel, Kornelia

doi:10.1002/chin.199046360

Cited by 40 publications

(56 citation statements)

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“…Penicillin G was kindly provided by Antibióticos S.A. (León, Spain). PEI (25,60, and 600 kDa), streptomycin sulfate, and isopropyl-␤-D-thiogalactopyranoside (IPTG) were supplied by Sigma-Aldrich (St. Louis, MO). DEAE-Sepharose and cyanogen bromide (CNBr)-activated Sepharose 4B were purchased from Amersham Biosciences (Uppsala, Sweden).…”

Section: Methodsmentioning

confidence: 99%

“…Nevertheless, the main drawback of the ionic immobilization strategy is the desorption of the proteins during operation, promoting biocatalyst inactivation and product contamination (25,44). The use of ionic polymers coating the surface of solid supports (e.g., polyethyleneimine [PEI] or dextran sulfate) not only provides a stronger adsorption but also prevents the alteration of the three-dimensional protein structure since the polymer becomes adapted to the enzyme instead of forcing the protein to become adapted to the support (13,31).…”

mentioning

confidence: 99%

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Genetic Modification of the Penicillin G Acylase Surface To Improve Its Reversible Immobilization on Ionic Exchangers

Montes

Grazú

López‐Gallego

et al. 2007

Appl Environ Microbiol

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A new mutant of the industrial enzyme penicillin G acylase (PGA) from Escherichia coli has been designed to improve its reversible immobilization on anionic exchangers (DEAE-or polyethyleneimine [PEI]-coated agarose) by assembling eight new glutamic residues distributed homogeneously through the enzyme surface via site-directed mutagenesis. The mutant PGA is produced and processed in vivo as is the native enzyme. Moreover, it has a similar specific activity to and shows the same pH activity profile as native PGA; however, its isoelectric point decreased from 6.4 to 4.3. Although the new enzyme is adsorbed on both supports, the adsorption was even stronger when supports were coated with PEI, allowing us to improve the enzyme stability in organic cosolvents. The use of restrictive conditions during the enzyme adsorption on anionic exchangers (pH 5 and high ionic strength) permitted us to still further increase the strength of adsorption and the enzyme stability in the presence of organic solvents, suggesting that these conditions allow the penetration of the enzyme inside the polymeric beds, thus becoming fully covered with the polymer. After the enzyme inactivation, it can be desorbed to reuse the support. The possibility to improve the immobilization properties on an enzyme by site-directed mutagenesis of its surface opens a promising new scenario for enzyme engineering.

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Genetic Modification of the Penicillin G Acylase Surface To Improve Its Reversible Immobilization on Ionic Exchangers

Montes

Grazú

López‐Gallego

et al. 2007

Appl Environ Microbiol

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“…However, most protocols have some drawbacks with immobilization under mild experimental conditions. A major one is the requirement for large amounts of protein per milliliter of support throughout long-term handling of the activated supports when the immobilization is carried out at an industrial level 14 .…”

mentioning

confidence: 99%

Purification of bovine serum paraoxonase and its immobilization on Eupergit C 250 L by covalent attachment

Sayın

Guler

2014

Journal of Enzyme Inhibition and Medicinal Chemistry

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Serum paraoxonase (PON1) is a high-density lipoprotein (HDL)-associated enzyme that protects lipoproteins, both low-density lipoprotein (LDL) and HDL, against oxidation, and is considered as an antioxidative/anti-inflammatory component of HDL. In this study, PON1 was purified from bovine serum by ammonium sulfate precipitation and hydrophobic interaction chromatography on sepharose-4B-L-tyrosine-1-napthylamine. It was then immobilized on an unmodified Eupergit Õ C 250 L support. The immobilized PON1 retained a high catalytic activity and showed increased thermal stability compared to the native enzyme.

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“…It has many well known advantages, all affecting the production cost (Çetinus and Nursevin Öztop, 2003;Choi and Yiu, 2004;Liu et al, 2009). With immobilized or adsorbed enzymes, improved stability, reusability, continuous operation, possibility of better control of reactions, high purity and product yields and hence, more favorable economic factors can be expected (Kennedy and Melo, 1990). In this study, the adsorption, electrokinetic properties of biomolecules such as casein and catalase onto sepiolite surface were studied as a function of incubation time, concentrations of biomolecules, pH of the solution and temperature.…”

mentioning

confidence: 99%

Thermodynamics, Kinetics and Adsorption Properties of Some Biomolecules onto Mineral Surfaces

Demirbaş¹,

Alkan²

2011

Thermodynamics - Kinetics of Dynamic Systems

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ChemInform Abstract: Immobilized Enzymes and Cells

Cited by 40 publications

References 0 publications

Genetic Modification of the Penicillin G Acylase Surface To Improve Its Reversible Immobilization on Ionic Exchangers

Genetic Modification of the Penicillin G Acylase Surface To Improve Its Reversible Immobilization on Ionic Exchangers

Purification of bovine serum paraoxonase and its immobilization on Eupergit C 250 L by covalent attachment

Thermodynamics, Kinetics and Adsorption Properties of Some Biomolecules onto Mineral Surfaces

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