Affinity interaction of hydroxypyruvate reductase from Methylophilus spp. with Cibacron blue F3GA-derived poly(HEMA EGDMA) microspheres: partial purification and characterization
Yükleniyor...
Tarih
1999
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Elsevier Sci Ltd
Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
A methylotrophic hydroxypyruvate reductase was partially purified and characterized from Methylophilus spp. using the biomimetic dye, Cibacron Blue F3FA attached to poly(HEMA-EGDMA) microspheres. The absorption capacities of the dye-affinity microspheres were determined by changing pH and the concentration of the proteins in the adsorption medium. Hydroxypyruvate reductase was desorbed from the dye-affinity support specifically with 2 mM NADH solution. The enzyme was purified 10.4-fold with 47% yield. The molecular mass and subunit molecular mass of the enzyme was estimated to be 75 kDa and 37 kDa on the basis of its mobility in polyacrylamide and SDS-polyacrylamide gels, respectively. This suggested a homogeneous dimer structure. The optimal pH was between 5.0 and 7.0, and the maximum enzyme activity was obtained at 50 degrees C. The K-m values of hydroxpyruvate reductase were 0.222 mM for hydroxpyruvate and 0.067 mM for NADH. (C) 1999 Elsevier Science Ltd. All rights reserved.
Açıklama
Anahtar Kelimeler
hydroxypyruvate reductase, NADH dependent enzyme, Methylophilus spp., affinity adsorption, poly(HEMA-EGDMA) microspheres, Cibacron Blue F3GA
Kaynak
Process Biochemistry
WoS Q Değeri
Q1
Scopus Q Değeri
Q1
Cilt
34
Sayı
4
Künye
closedAccess
