Covalent and Metal-Chelate Immobilization of a Modified 2-Haloacid Dehalogenase for the Enzymatic Resolution of Optically Active Chloropropionic Acid

Ordaz, Enrique; Garrido-Pertierra, Amando; Gallego, Manuel Requena; Puyet, Antonio

doi:10.1021/bp990134k

Cited by 25 publications

(8 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…with a peptide tail containing six histidines and overexpressed the enzyme in E. coli. 58 The His-tagged protein was purified by single-step affinity chromatography on Zn 2+ -chelating sepharose. The enzyme was immobilized on chelating sepharose and EUPERGIT C, respectively (immobilized activities not reported).…”

Section: D Burkholderia Cepaciamentioning

confidence: 99%

EUPERGIT Oxirane Acrylic Beads: How to Make Enzymes Fit for Biocatalysis

Böller

Meier

Menzler

2002

Org. Process Res. Dev.

163

View full text Add to dashboard Cite

Enzyme recycling is essential for the development of large-scale enzyme-catalyzed biotransformations. Recycling is most convenient using enzymes immobilized on solid supports. Although immobilization on solid supports has been pursued since the 1950s, there are no general rules for selecting the best support for a given application. The commercial products EUPERGIT C and EUPERGIT C 250 L have been used for a wide variety of different enzymes and reactions. The present review draws up a comprehensive application profile of both EUPERGIT carriers. The reader gets (a) examples of biotransformations using oxidoreductases, transferases, hydrolases and lyases immobilized on EUPERGIT; (b) key data of the biotransformations, i.e., scale, yield, purity, and enantiomeric excess; (c) efficiency of the immobilization (% immobilized activity); (d) where appropriate, operational stability of the immobilized enzyme preparations, that is, number of cycles, residual activity; (e) specific advantages of the immobilized enzyme over the free enzyme apart from enzyme recycling, for example, improved stability and selectivity. Thus, the present review can serve as a guideline when selecting a resin for enzyme immobilization. Literature published between 1985 and 2000 is covered.

show abstract

Section: D Burkholderia Cepaciamentioning

confidence: 99%

EUPERGIT Oxirane Acrylic Beads: How to Make Enzymes Fit for Biocatalysis

Böller

Meier

Menzler

2002

Org. Process Res. Dev.

163

View full text Add to dashboard Cite

show abstract

“…IMAC applications have been used in separation of amino acids, nucleotides, and small molecules such as amines and in purification of various proteins (Berna et al., 1997; Liu and Yu, 1990; Sulkowski, 1989). IMAC is also used to determine surface topography of proteins (Boden et al., 1998) and offers many advantages compared with chromatographic methods such as high binding capacity and high recovery yield, nondenaturing elution conditions beside cheapness and stability (Brena et al., 1994; Ordaz et al., 2000).…”

Section: Introductionmentioning

confidence: 99%

Poly-(l)-histidine immobilized cryogels for lysozyme purification

Çimen

Türkmen

Deni̇zli̇

2016

Adsorption Science & Technology

View full text Add to dashboard Cite

Immobilized metal ion affinity chromatography is one of the methods used for the adsorption of proteins. In this study, poly(glycidyl methacrylate) cryogel discs were prepared by free radical polymerization. The metal chelating groups were polymeric chain of poly-(L)-histidine (mol wt ! 5000) having poly-imidazole ring sequence. Then, Cu(II), Zn(II), and Ni(II) ions were separately chelated on the poly-(L)-histidine immobilized poly(glycidyl methacrylate) cryogel discs to be used in immobilized metal ion affinity chromatography separation of lysozyme. The swelling test, Fourier transform infrared spectroscopy, Brunauer-Emmett-Teller, and scanning electron microscopy were performed to characterize both poly(glycidyl methacrylate) and poly-(L)-histidine immobilized poly(glycidyl methacrylate) cryogel discs. The effects of the pH, lysozyme concentration, adsorption time, and ionic strength on the adsorption capacity were studied. These parameters were varied between 4.0 and 8.0 for pH, 0.0 and 2.0 mg/ml for initial lysozyme concentration, 0 and 120 min for adsorption time, and 0.0 and 1.0 mM for ionic strength. The maximum lysozyme adsorption capacity of the Cu(II), Zn(II), and Ni(II) ions chelated poly-(L)-histidine immobilized poly(glycidyl methacrylate) cryogel discs was 36.4, 26.8, and 17.3 mg/g cryogel, respectively. Desorption of lysozyme from cryogel discs was easily achieved by 1.0 M NaCI solution. Repeated adsorption-elution processes showed that these cryogel discs were suitable for repeatable lysozyme adsorption. Adsorption isotherms fitted to Langmuir model and adsorption kinetics suited to pseudo-second order model. Thermodynamic parameters (i.e. DH , DS , DG) were also calculated from Langmuir isotherms at different temperatures.

show abstract

“…In addition to protein purification, immobilized metal affinity adsorbents have also been used as the matrices for a wide variety of applications, including enzyme immobilization [5,6], protein refolding [7,8], and protein reconstitution [9]. For example, in one of our recent studies, an integrated enzyme purification and immobilization process for biotransformation with a silica-based immobilized metal affinity adsorbent has been demonstrated [10].…”

Section: Introductionmentioning

confidence: 99%