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Hi,
I have another question too. I am working on Cellulases from Bacteria. Is it possible that i centrifuge the culture medium in eppendorphs to get the crude enzyme but use this crude enzyme next day to perform assay for different parameters in its charaterization? If i keep this crude enzyme in refrigerator for a night, will it change the nature of the enzyme?
San.
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Conversion of Absorbance into enzyme units
05 August 2012 - 06:49 AM
Hi all,
I'm working on cellulases from bacteria. I've taken O.D after several parameters to check the enzyme activity. I'm using Sodium Acetate buffer of pH 5 (0.1 M) in my enzyme assay. I take 0.2 ml crude enzyme, 0.5 ml substrate buffer (substrate prepared in Acetate buffer of pH 5 (0.1M)) ad 0.3 ml of Acetate buffer as stated above. It makes up the reaction mixture 1 ml in total. I incubate it at 50 C for 15 mins, then add 3 ml DNS and then boil for 5 mins, cool down and take the Absorbance. For example, if Spec give the reading as 0.375. How would i convert it into enzyme units?
I have tried to search it on internet but couldnt find anything understandable.
Thanks in advance
San.
I'm working on cellulases from bacteria. I've taken O.D after several parameters to check the enzyme activity. I'm using Sodium Acetate buffer of pH 5 (0.1 M) in my enzyme assay. I take 0.2 ml crude enzyme, 0.5 ml substrate buffer (substrate prepared in Acetate buffer of pH 5 (0.1M)) ad 0.3 ml of Acetate buffer as stated above. It makes up the reaction mixture 1 ml in total. I incubate it at 50 C for 15 mins, then add 3 ml DNS and then boil for 5 mins, cool down and take the Absorbance. For example, if Spec give the reading as 0.375. How would i convert it into enzyme units?
I have tried to search it on internet but couldnt find anything understandable.
Thanks in advance
San.
