Premium
PURIFICATION AND PROPERTIES OF GLUCOSE 6‐PHOSPHATE DEHYDROGENASE FROM CORIANDER ( CORIANDRUM SATIVUM ) LEAVES
Author(s) -
DEMIR HÜLYA,
BEYDEMIR SÜKRÜ,
ÇIFTÇI MEHMET,
KÜFREVIOĞLU Ö. ÇRFAN
Publication year - 2004
Publication title -
journal of food biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.507
H-Index - 47
eISSN - 1745-4514
pISSN - 0145-8884
DOI - 10.1111/j.1745-4514.2004.tb00062.x
Subject(s) - coriandrum , chemistry , chromatography , sephadex , dehydrogenase , size exclusion chromatography , enzyme , glucose 6 phosphate dehydrogenase , phosphate , molecular mass , enzyme assay , fractionation , potassium phosphate , sativum , biochemistry , biology , botany
Glucose‐6‐phosphate dehydrogenase (D‐glucose‐6‐phosphate: NADP + oxidoreductase, EC 1.1.1.49; G6PD) was purified from coriander (Coriandrum sativum) leaves; the kinetic behavior and some properties of the enzyme were also investigated. The purification was done at 4C and involved two steps: ammonium sulfate fractionation, and DEAE‐Sephadex A50 ion exchange chromatography. The enzyme was obtained with a yield of 26.4% and had a specific activity of 1.826 U/mg protein. Optimum pH, stable pH, optimum temperature, molecular weight, K M and V max values for NADP + and glucose 6‐phosphate (G6‐P) were also determined.The overall purification was about 74‐fold. SDS‐PAGE of the purified enzyme showed a single band. Enzymatic activity was spectrophotometrically measured according to Beutler's method at 340 nm.The molecular mass was estimated to be 74.4 kDa by SDS‐PAGE and 73.2 kDa by Sephadex G‐200 gel filtration column chromatography. The enzyme had an optimum pH at 8.5 and was stable at pH 8.0 in 0.1 M Tris‐HCl buffer. The optimum temperature was at 30C. The K M values for NADP + and G6‐P were 0.026 mM and 0.116 mM, respectively. The V max values for these substrates were 0.035 EU/mL and 0.038 EU/mL, respectively.