Purification and Characterization of the Glucose Oxidase from Penicillium notatum

Abstract

This study aims to purification and characterization of the glucose oxidase enzyme from Penicillium notatum, the enzyme was purified by ammonium sulfate precipitation (60%), dialysis and gel filtration chromatography using sephadex G-200, A trial for the purification of glucose oxidase using gel filtration technique resulted in one type of glucose oxidase with specific activity of (62.382 U/mg) with (7.385 folds) purification. the purified glucose oxidase had a maximum activity at pH = 5.5, 45 °C, glucose oxidase was stable with pH values ranging between (5 – 6) and the enzyme was maintained the activity when it incubated into (25 -35) °C for 15 minutes, analyses of the glucose oxidase for molecular weight was carried out by PAGE and SDS-PAGE electrophoresis, which revealed 78 KDa, also molecular weight of the glucose oxidase was achieved by gel filtration technique and was found 87 KDa this means that enzyme consisting of only one subunit, the Km and Vmax value of glucoamylase (B) were (19.6 mM, 7.5 mM/min ) respectively using different concentration of glucose.

Characterization of glucose Oxidase from Penicillium chrysogenum MDC 8358: Prospects for application in food industry

“…Aspergillus tubingensis [20], Penicillium amagasakiense [21], Penicillium notatum [1,[22][23], Penicillium variabile P16 [24], Penicillium funiculosum [25][26], Penicillium sp.…”

Enzyme production ofd-gluconic acid and glucose oxidase: successful tales of cascade reactions

“…30,32 For commercial applications, glucose oxidase has been produced and purified from fungi, especially from Aspergillus and Penicillium species. 19,[33][34][35][36][37][38][39][40][41][42][43][44][45] The great success of these two species is mainly due to their metabolic versatility and their recognition as GRAS (generally recognized as safe). 19,38 Among the Aspergillus species, remarkable attention has been given to A. niger over the last 60 years.…”

“…84 In general, fungal glucose oxidases have an isoelectric point in the range of pH 4.0-5.0, 16 optimal activity in the pH range from 4.0 to 7.0, 14,15,29,31,33,40,75,84 and optimal temperature in the range of 40-60°C. 15,29,33,40,75,84 Glucose oxidase from A. niger does not require metal ions to express its activity; however, it is inhibited by Ag + , Hg 2+ , Cu 2+ , Mg 2+ and Ca 2+ . 19 Very importantly in the context of this review, hydrogen peroxide is a strong deactivating agent of glucose oxidase.…”

Development of an Effective and Economic Biosensor for Diabetic Blood Monitoring Based on MWCNTs, Artificial Redox Mediator Ferrocene, Nafion Polymer and a Local Extracted and Purified Glucose Oxidase Enzyme from Penicillium Notatum F-158 Fungus

“…Purification of glucose oxidase enzyme.-The extracted enzyme was then purified using ammonium sulfate precipitation, dialysis and gel filtration chromatography 29 as described in the (ESM).…”

“…28 One enzyme activity unit (U) was defined as the amount of enzyme releasing 1 μmol of H 2 O 2 from the substrate in 1 min under standard assay conditions. 29 In this assay, glucose was catalyzed by extracted GOx and converted to gluconic acid and H 2 O 2 . The formed H 2 O 2 reacted with 4-aminoantipyrine and phenol, producing a red color antipyrilquinoneimine dye in the presence of horseradish peroxidase (HRP) according to the following reactions: 28 β δ…”