Quantitativly Measuring Enzyme Activity?

[deleted-141596]

For my 2015-2016 project, I will be doing a continuation of my 2014-15 project "Using sugar polymers for plastic alternatives". In the new project I will be trying to improve the sugar bioplastics I made last year by adding different types of minerals/substances that premote enzyme activity (These additives are my independent variable). I will put each sugar bioplastic in a test tube with nutrient broth, bacillus subtilis and an additive. My goal is that these additives will premote more enzyme activity and make the plastics breakdown faster.

My problem is that I am having a hard time finding a way to quantitativly measure the activity of the enzymes. I found that I can use assays for the enzymes I am looking at, however one of the assays (the agarase one) would be too dangerous by ISEF standards. Is there another method I could use? I am looking for α-amylase, laccase, and agarase activity. Thank you for your help in advance!

[deleted-143835]

Hi Danni,

What an interesting idea! It's great to hear that you're continuing to research a topic you've already explored and enjoyed.
One idea off the top of my head for measuring enzyme activity would be to use a UV-Visible Absorption Spectrophotometer. The UV-vis allows you to "watch" enzyme activity by measuring the change in light absorption of the solution as the enzyme catalyzes the reaction. A UV-vis is simple to use, and its graphs are easy to interpret, so this could be an ideal option for you; a good next step would be to see if the UV-vis is "compatible" with those 3 enzymes. However, you would need to have access to this equipment, most often found in university labs. If you were planning on doing an agarase assay, I suppose you would need a lab anyway--please let me know your situation in regards to equipment, and perhaps we can figure out a home test if facilities are an issue!
I'm also curious as to how long these reactions are--that might help you find a good assay to use.

Hope this helps! Please feel free to post back with more questions.

Thanks,
scibuddyAK

[deleted-141596]

scibuddyAK,
That's a great idea! From what I've found, the spectrophotometer method should work with all of the enzymes I'm testing. I was also planning on getting lab space at a local college, so this is definitely a viable method for my project. I may post back with more questions as I do more research. Thank you for your help!

[deleted-143835]

Hi Danni,

So glad to hear that helped! Yes, please feel free to post back with any more questions you may have. Good luck, and let me know how it goes!

Thanks,
scibuddyAK