WHS Advanced Chemistry Mr Spangler Mycro Remediation How

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WHS Advanced Chemistry Mr. Spangler Mycro Remediation How the production of magnizeperaxadase changes over

WHS Advanced Chemistry Mr. Spangler Mycro Remediation How the production of magnizeperaxadase changes over time Research Question: How does the production of lignin degrading enzymes in pleurotus ostreatus (manganese peroxidase) change over time? Purpose: To help establish a more uniform Xander Burger Hypothesis: The appearance of the manganese peroxidase enzyme in pleurotus ostreatus will follow a curved distribution skewed to the right. The pleurotus will produce manganese peroxidase quickly at the start to break down the substrate as food then slowly produce less as the substrate begins to be consumed. method of mycoremediation that can be followed and reproduced so that the process is more fluid and efficient. Method: I will cultivate pearl oyster mushrooms in a sealed bags with coffee grounds as my Background: My interest in this project The assay: The assay is performed by adding a extract of culture to a buffer solutions with H 2 O 2 and Manganese then put into a spectrophotometer in order to tell the concentration. When oxidised by the peroxidase enzyme, the phenolic dye (phenol red) in the buffer solution, changes color. The concentration of the solution is proportional to the wavelength of the color change (the amount of oxidized phenol red). comes from the incredible and mysterious world of the fungi. Fungi are the foundation of all ecological systems. They are such a key factor for life on earth that some have even said that “fungi cultivated us. ” My specific interest is in the powerful enzyme mechanism that fungi use to rot or breakdown carbon based molecules and use for food. Fungus like the pearl oyster or pleurotus ostreatus, is known for its ability to breakdown tough hydrocarbons. It has been used on oil and chemical spills to breakdown toxins and remediate the environment. The pleurotus is a white rot fungus which means that it consumes the lignin in wood, leaving the wood a lighter color. It uses powerful enzymes to do so. The pleurotus produce an enzyme called manganese peroxidase which is particularly good at removing chlorine atoms from carbon chains like that of aminopyralid (a common herbicide), the reason it is difficult to break down. This makes it a favorite for micro-remediators and make me wonder what else this fungus can do? I am particularly interested in how the appearance of these enzymes changes over time and in the future, how this affect the degradation of hydrocarbons in soil. substrate and a grain spawn for my culture. I will perform an enzyme assay every week for 3 week and see how the concentration changes over time. Spectrophotometer: The spectrophotometer quantitatively measures a material's reflected wavelength. It allows you determine the concentration of a substance based on color. Data: The graph below shows the test results of the pleurotus osteaus from its spawn bag (earliest stage of growth). It has a sharp drop at 600 nm at reaches zero at 620 nm. The absorption at 610 nm is 0. 37. The absorption of light at 610 nm is what determines the concentration of manganese peroxidase. Therefore, sense the abortion is low, 0. 37, there is a low concentration of the enzyme which is to be expected at this early stage of growth. In the second week the absorption almost doubled, jumping to. 62 then drops down to 0 in the third week representing no production of manganese peroxidase. What to Improve on: Sense this was my first time Conclusion: I don’t think I can make any definitive conclusions with my data because of how little I have of it. That said, my data follows the pattern that I predicted in my hypothesis to an extent. It goes up maxing out at week two 0. 62 then goes down in week 3 roughly following a bell curve. From this data I would say that the prime time period to plant for remediation is at 2 weeks but further testing is needed. performing this test I think if I had a few more weeks I could more consistently perform the test. I would also order my mushrooms early and grow them in a more uniform and consistent way. What’s Next? I would love to expand this project by using samples in actual mycro remediation and a bioassay to assess the effectiveness and optimal time of application. Special Thanks to Cliff Bradley- Montana Bio Agriculture References Hofrichter, Martin. “Review: Lignin Conversion by Manganese Peroxidase (Mn. P). ” Science Direct , 2002, www. sciencedirect. com/science/article/pii/S 0141022901005282 “Using Fungi To Clean Up Pollutants. ” Permaculture Magazine, 2 Nov. 2018, www. permaculture. co. uk/readers-solutions/using-fungi-cleanpollutants. . The 4 solutions used in enzyme assay Spectrophotometer results from initial test