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Purification and investigation of some kinetic properties of glucose-6-phosphate dehydrogenase from parsley (Petroselinum hortense) leaves.
Prep Biochem Biotechnol. 2002 May; 32(2):173-87.PB

Abstract

In this study, glucose-6-phosphate dehydrogenase (D-glucose-6-phosphate: NADP+ oxidoreductase, EC 1.1.1.49; G6PD) was purified from parsley (Petroselinum hortense) leaves, and analysis of the kinetic behavior and some properties of the enzyme were investigated. The purification consisted of three steps: preparation of homogenate, ammonium sulfate fractionation, and DEAE-Sephadex A50 ion exchange chromatography. The enzyme was obtained with a yield of 8.79% and had a specific activity of 2.146 U (mg protein)(-1). The overall purification was about 58-fold. Temperature of +4 degrees C was maintained during the purification process. Enzyme activity was spectrophotometrically measured according to the Beutler method, at 340 nm. In order to control the purification of enzyme, SDS-polyacrylamide gel electrophoresis was carried out in 4% and 10% acrylamide for stacking and running gel, respectively. SDS-polyacrylamide gel electrophoresis showed a single band for enzyme. The molecular weight was found to be 77.6 kDa by Sephadex G-150 gel filtration chromatography. A protein band corresponding to a molecular weight of 79.3 kDa was obtained on SDS-polyacrylamide gel electrophoresis. For the enzymes, the stable pH, optimum pH, and optimum temperature were found to be 6.0, 8.0, and 60 degrees C, respectively. Moreover, KM and Vmax values for NADP+ and G6-P at optimum pH and 25 degrees C were determined by means of Lineweaver-Burk graphs. Additionally, effects of streptomycin sulfate and tetracycline antibiotics were investigated for the enzyme activity of glucose-6-phosphate dehydrogenase in vitro.

Authors+Show Affiliations

Faculty of Erzincan Education, Atatürk University, Erzurum, Turkey.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

12071647

Citation

Coban, T Abdül Kadir, et al. "Purification and Investigation of some Kinetic Properties of Glucose-6-phosphate Dehydrogenase From Parsley (Petroselinum Hortense) Leaves." Preparative Biochemistry & Biotechnology, vol. 32, no. 2, 2002, pp. 173-87.
Coban TA, Ciftçi M, Küfrevioğlu OI. Purification and investigation of some kinetic properties of glucose-6-phosphate dehydrogenase from parsley (Petroselinum hortense) leaves. Prep Biochem Biotechnol. 2002;32(2):173-87.
Coban, T. A., Ciftçi, M., & Küfrevioğlu, O. I. (2002). Purification and investigation of some kinetic properties of glucose-6-phosphate dehydrogenase from parsley (Petroselinum hortense) leaves. Preparative Biochemistry & Biotechnology, 32(2), 173-87.
Coban TA, Ciftçi M, Küfrevioğlu OI. Purification and Investigation of some Kinetic Properties of Glucose-6-phosphate Dehydrogenase From Parsley (Petroselinum Hortense) Leaves. Prep Biochem Biotechnol. 2002;32(2):173-87. PubMed PMID: 12071647.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Purification and investigation of some kinetic properties of glucose-6-phosphate dehydrogenase from parsley (Petroselinum hortense) leaves. AU - Coban,T Abdül Kadir, AU - Ciftçi,Mehmet, AU - Küfrevioğlu,O Irfan, PY - 2002/6/20/pubmed PY - 2002/12/27/medline PY - 2002/6/20/entrez SP - 173 EP - 87 JF - Preparative biochemistry & biotechnology JO - Prep Biochem Biotechnol VL - 32 IS - 2 N2 - In this study, glucose-6-phosphate dehydrogenase (D-glucose-6-phosphate: NADP+ oxidoreductase, EC 1.1.1.49; G6PD) was purified from parsley (Petroselinum hortense) leaves, and analysis of the kinetic behavior and some properties of the enzyme were investigated. The purification consisted of three steps: preparation of homogenate, ammonium sulfate fractionation, and DEAE-Sephadex A50 ion exchange chromatography. The enzyme was obtained with a yield of 8.79% and had a specific activity of 2.146 U (mg protein)(-1). The overall purification was about 58-fold. Temperature of +4 degrees C was maintained during the purification process. Enzyme activity was spectrophotometrically measured according to the Beutler method, at 340 nm. In order to control the purification of enzyme, SDS-polyacrylamide gel electrophoresis was carried out in 4% and 10% acrylamide for stacking and running gel, respectively. SDS-polyacrylamide gel electrophoresis showed a single band for enzyme. The molecular weight was found to be 77.6 kDa by Sephadex G-150 gel filtration chromatography. A protein band corresponding to a molecular weight of 79.3 kDa was obtained on SDS-polyacrylamide gel electrophoresis. For the enzymes, the stable pH, optimum pH, and optimum temperature were found to be 6.0, 8.0, and 60 degrees C, respectively. Moreover, KM and Vmax values for NADP+ and G6-P at optimum pH and 25 degrees C were determined by means of Lineweaver-Burk graphs. Additionally, effects of streptomycin sulfate and tetracycline antibiotics were investigated for the enzyme activity of glucose-6-phosphate dehydrogenase in vitro. SN - 1082-6068 UR - https://www.unboundmedicine.com/medline/citation/12071647/Purification_and_investigation_of_some_kinetic_properties_of_glucose_6_phosphate_dehydrogenase_from_parsley__Petroselinum_hortense__leaves_ L2 - https://www.tandfonline.com/doi/full/10.1081/PB-120004129 DB - PRIME DP - Unbound Medicine ER -