Biochemistry Question

[Missserica]

Hello there,

I have this problem from my biochemistry class that I need some help with. Hopefully you can help me .. I feel like this is not a hard question, it's just worded weird and I don't know how to answer it.

Enzyme X is a highly pigmented protein that imparts the characteristic color to certain blue-green algae. It also facilitates a reaction necessary to the survival of this species; we can follow the progress of this reaction by measuring the conversion of Substance X to Substance Y.

1. Given a pure preparation of these algae, and the required supplies and equipment, devise and outline an empirical procedure for purifying Enzyme X.

Photosynthesis is not involved in the answer..

Ok, any help would be mucho appreciated.

Thanks.

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[captaintripps]

Hello there,

I have this problem from my biochemistry class that I need some help with. Hopefully you can help me .. I feel like this is not a hard question, it's just worded weird and I don't know how to answer it.

Enzyme X is a highly pigmented protein that imparts the characteristic color to certain blue-green algae. It also facilitates a reaction necessary to the survival of this species; we can follow the progress of this reaction by measuring the conversion of Substance X to Substance Y.

1. Given a pure preparation of these algae, and the required supplies and equipment, devise and outline an empirical procedure for purifying Enzyme X.

Photosynthesis is not involved in the answer..

Ok, any help would be mucho appreciated.

Thanks.

I think this question is an exercise in understanding some methods of protein purification which may have been discussed in your class. The protein/enzyme has 2 useful properties that can be exploited in the process of purifying; it is pigmented so it can be visualized during procedures (indicates presence of protein in the fraction) and the reaction catalyzed can be monitored as indicated X>Y can be measured (indicates that protein is still native). So using conventional techniques of biochemical purification - cell fractionation, size exclusion chromatography, affinity chromatography etc., and then protein analysis by polyacrylamide electrophoresis and coomassie staining a purified protein can be obtained in theory. The pigment and reaction allow two distinct ways to monitor the presence of the protein during purification.
Hope this helps, just review some purification methods in a biochem book and ask if there is an antibody available, this will be useful also.