I am trying to extract glucose oxidase from used mycelium of Aspergillus niger after producing sodium gluconate. How do I get glucose oxidase from it?
The typical glucose oxidase assay involves the presence of peroxidase and a chromogenic substrate (i.e. ABTS). In this way, the hydrogen peroxide produced by GOX is used by the second enzyme to convert ABTS to its oxidized form, presenting a strong absorbtion around 420 nm.
I did as Paolo Zucca mentioned and worked super. Here are the conditions:
The reaction mixture in total volume of 1 ml consisted of 0.1 M sodium phosphate pH 7.0, 5 mM 2,2´-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), 0.15 U horseradish peroxidise, 1 mM potassium hexacyanoferrate(III) (K3[Fe(CN)6]) and 10 U glucose oxidase. The oxidation of ABTS was followed at 725 nm (ε = 19 mM-1cm-1). K3[Fe(CN)6] was used to oxidize the reducing agent cysteine of the medium, which bleached the blue-green oxidized ABTS (ABTS+) (Bergmeyer and Bernt, 1974)
I usually use the coupled enzymatic assay with peroxidase and the chromogen o-dianisidine for estimating glucose oxidase activity and measure it in an spectrophotometer. This methodology is suitable also for microplates and it is a relative low cost assay. If you want the complete protocol, you can contact me by e-mail or by direct message.
The glucose oxidase enzyme (GOx) is an oxido-reductase that catalyzes the oxidation of glucose to hydrogen peroxide and D-glucono-δ-lactone. In cells, it aids in breaking the sugar down into its metabolites. The Glucose Oxidase in sample will recognize d-glucose as a specific substrate leading to proportional color development. The activity of GOx can be easily quantified colorimetrically (λ = 570 nm) or fluorometrically (Ex/Em = 535/587). This assay detects GOx activity as low as 0.01mU with our unit definition.
Hi Ivana Djurdjevic
Could you send me your detailed protocol? my email is: [email protected]
Hello Carlos Diaz-Tufinio
Could you please send me the complete protocol for detection of glucose oxidase.
My e-mail id is [email protected]
Hi Ivana Djurdjevic can you send me the detailed procedure
How to estimate glucose oxidase activity?. Available from: https://www.researchgate.net/post/How_to_estimate_glucose_oxidase_activity [accessed Mar 24, 2015].
Hello Ivana Dejurdjevic
could you please send me your protocol
my e-mail is [email protected]
Hello Ivana Djurdjevic,
Please also send me the detailed procedure and protocol. Will be very grateful for your help.
email: [email protected]
Hello Ivana Djurdjevic,
Could you please also send me the detailed procedure and protocol. Will be very grateful for your help.
email: [email protected]
Hello Carlos Diaz-Tufinio
Could you please send me the detailed protocol for coupled enzymatic assay for detection of glucose oxidase.
my mail-id : [email protected]
Hi Ivana Djurdjevic, could you please send the detailed protocol for glucose oxidase assay. My email address is [email protected]. Thanks.
could you please send the detailed protocol for glucose oxidase assay. My email address is [email protected]. Thanks.
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