Heterofunctional magnetic metal-chelate-epoxy supports for the purification and covalent immobilization of benzoylformate decarboxylase from pseudomonas putida and its carboligation reactivit
Yükleniyor...
Tarih
2015
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Wiley
Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
In this study, the combined use of the selectivity of metal chelate affinity chromatography with the capacity of epoxy supports to immobilize poly-His-tagged recombinant benzoylformate decarboxylase from Pseudomonas putida (BFD, E.C. 4.1.1.7) via covalent attachment is shown. This was achieved by designing tailor-made magnetic chelate-epoxy supports. In order to selectively adsorb and then covalently immobilize the poly-His-tagged BFD, the epoxy groups (300 mu mol epoxy groups/g support) and a very small density of Co2+-chelate groups (38 mu mol Co2+/g support) was introduced onto magnetic supports. That is, it was possible to accomplish, in a simple manner, the purification and covalent immobilization of a histidine-tagged recombinant BFD. The magnetically responsive biocatalyst was tested to catalyze the carboligation reactions. The benzoin condensation reactions were performed with this simple and convenient heterogeneous biocatalyst and were comparable to that of a free-enzyme-catalyzed reaction. The enantiomeric excess (ee) of (R)-benzoin was obtained at 99 +/- 2% for the free enzyme and 96 +/- 3% for the immobilized enzyme. To test the stability of the covalently immobilized enzyme, the immobilized enzyme was reused in five reaction cycles for the formation of chiral 2-hydroxypropiophenone (2-HPP) from benzaldehyde and acetaldehyde, and it retained 96% of its original activity after five reaction cycles. Chirality 27:635-642, 2015. (c) 2015 Wiley Periodicals, Inc.
Açıklama
Anahtar Kelimeler
Immobilized benzoylformate decarboxylase, Metal-chelate-epoxy magnetic support, Metal-affinity magnetic nanoparticles, Selective immobilization, Covalent attachment, One step purification
Kaynak
Chirality
WoS Q Değeri
Q2
Scopus Q Değeri
Q2
Cilt
27
Sayı
9
