EXPERIMENTS TO EXTRACT PROTEASE FROM FRUIT AND DETERMINE PROTEIN CONCENTRATION IN MUSKMELON
There are some word document that need to be emailed to you as research paper instruction material. you will need to used all the documents attach as guideline. please organise it as: Introduction, method(including calculations), results, discussion.
AIM
1. Set up a protease assay using a known protease sample (Neutrase ) and a previously published method.
2. Assay a cell free extract from a fruit sample for protease activity using pineapple as a control.
3. Calculate yield and specific activity for this sample.
4. You will investigate, by your own design, a suitable purification step or initial extraction method for this protease.
5. Analysis of your results.
Introduction 15% (IN LOGICAL FLUENT ORDER)
– Setting the scene and introducing the aim through coherent, relevant background information that quickly focuses on the specific topic and aim. Has to be clear and straight to the point. Reference scientific source used to find information.
– Focus on Protease Assays, Lowry assay, Purification methods for Muskmelon from public literature, protease in fruit; MUSKMELON (chosen fruit), pineapple (positive control)
– Talk about the protease enzyme itself and the enzyme reaction
– Talk about the purification methods used (include details of the protease assay
– Use primary source material, (theories, and literature) add any addition bits you think is relevant
– Work should be coherent and well-structured and meet aim and objective of the experiments.
Method (experimental design) 20% (IN LOGICAL FLUENT ORDER)
• Use appropriate detail and level of accuracy on design: one or two sentences.
• Make sure method is laid out so someone can follow and repeat
-You will be using pineapple as a positive control as the protease within this fruit is very well characterised and known to be very stable.
• Talk about how fruit extracts where prepared (fig 1)
• Talk about protease assay (2 Assays; standard assay, enzyme assay – Fig 2), (Volume of assay, concentration of reagents in assay, pH, temperature of assay, time; when and how is assay stopped, absorbance/wavelength; spectrophotometer, calculation of V0; as Abs/10mins)
• Talk about Lowry method (fig 3) (volumes, range of BSA, absorbance/wavelength from spectrophotometer)
• What is the function of 120mg/ml standard
• Talk about how to make the buffer (use buffer sheet and find optimum pH for buffer for this experiment from research)
Results 25% (IN LOGICAL COHERENT ORDER) ON EXCEL publication
• Use appropriate figures and tables and units to represent data and must have adequate detail of how data was produced and calculations achieved.
• Use High quality of text based description presented in logical order, showing clarity accuracy of data.
• For Lowry Protein determination results: table, figure (calibration curve graph to find unknown for dilutions, consider best way to plots data, whether to plot mean standard deviation/median/all points, find equation of line of best fit, gradient, R2 ), descriptive text to explain table and figures
• For Initial enzyme assay using Neutrase present: table and description text, find Absorbance change over time.
• For Fruit Extracts use: tables, Figure, descriptive text for table and figure
• For purification step use: table Figure and descriptive text table and figure
Calculations – useful equations:
Enzyme activity= total activity/ total volume (units/ml)
Specific enzyme activity = enzyme activity/protein concentration (units/mg)
Rate of reaction = change in absorbance/time
Discussion 30%
This should be extensive consideration and interpretation of the results. Must include significance and application of the results in wider context. Should not just be a description of the findings (which should be in the results). Results should be discussed with respect to relevant scientific resources (books/journal articles). Should not simply be a restating of the literature given in the introduction, but rather using the literature to explain (and possibly extend) specific aspects of the results. Should be reflective – consideration of the experimental design with suggestions for improvement for follows up experiments.
Presentation and referencing: ~ 10 %
Clear and concise scientific English with high standard of spelling and grammar. Correct referencing style
Example type sources to use: chitturi S 2013 studies on protein content protease activity anti oxidase potential and web sites like; United States department of agriculture.
THERE ARE SOME ATTACHMENTS THAT COMES WITH THIS WORK THAT IS ESSENTIAL AS GUILDLINE FOR THE WRITING.
PLEASE ensure you understand the nature of the laboratory research paper that is to be written. Please use the attached matterial i have provided fo guldlines to how the paper should be written and also using research sources for background knowlege of work and to support your work. Take into concideration the tables and Figures(graphs) that need to be made to represent results. furthermore, please take into consideration the results data and calculations that needs to provided in this work. i put trust in you that you will reflect this piece of work well. Any details or questions please feel free to contact me to ask. Many Thanks Kind Regards
There are some word document that need to be emailed to you as research paper instruction material. you will need to used all the documents attach as guideline. please organise it as: Introduction, method(including calculations), results, discussion.
AIM
1. Set up a protease assay using a known protease sample (Neutrase ) and a previously published method.
2. Assay a cell free extract from a fruit sample for protease activity using pineapple as a control.
3. Calculate yield and specific activity for this sample.
4. You will investigate, by your own design, a suitable purification step or initial extraction method for this protease.
5. Analysis of your results.
Introduction 15% (IN LOGICAL FLUENT ORDER)
– Setting the scene and introducing the aim through coherent, relevant background information that quickly focuses on the specific topic and aim. Has to be clear and straight to the point. Reference scientific source used to find information.
– Focus on Protease Assays, Lowry assay, Purification methods for Muskmelon from public literature, protease in fruit; MUSKMELON (chosen fruit), pineapple (positive control)
– Talk about the protease enzyme itself and the enzyme reaction
– Talk about the purification methods used (include details of the protease assay
– Use primary source material, (theories, and literature) add any addition bits you think is relevant
– Work should be coherent and well-structured and meet aim and objective of the experiments.
Method (experimental design) 20% (IN LOGICAL FLUENT ORDER)
• Use appropriate detail and level of accuracy on design: one or two sentences.
• Make sure method is laid out so someone can follow and repeat
-You will be using pineapple as a positive control as the protease within this fruit is very well characterised and known to be very stable.
• Talk about how fruit extracts where prepared (fig 1)
• Talk about protease assay (2 Assays; standard assay, enzyme assay – Fig 2), (Volume of assay, concentration of reagents in assay, pH, temperature of assay, time; when and how is assay stopped, absorbance/wavelength; spectrophotometer, calculation of V0; as Abs/10mins)
• Talk about Lowry method (fig 3) (volumes, range of BSA, absorbance/wavelength from spectrophotometer)
• What is the function of 120mg/ml standard
• Talk about how to make the buffer (use buffer sheet and find optimum pH for buffer for this experiment from research)
Results 25% (IN LOGICAL COHERENT ORDER) ON EXCEL publication
• Use appropriate figures and tables and units to represent data and must have adequate detail of how data was produced and calculations achieved.
• Use High quality of text based description presented in logical order, showing clarity accuracy of data.
• For Lowry Protein determination results: table, figure (calibration curve graph to find unknown for dilutions, consider best way to plots data, whether to plot mean standard deviation/median/all points, find equation of line of best fit, gradient, R2 ), descriptive text to explain table and figures
• For Initial enzyme assay using Neutrase present: table and description text, find Absorbance change over time.
• For Fruit Extracts use: tables, Figure, descriptive text for table and figure
• For purification step use: table Figure and descriptive text table and figure
Calculations – useful equations:
Enzyme activity= total activity/ total volume (units/ml)
Specific enzyme activity = enzyme activity/protein concentration (units/mg)
Rate of reaction = change in absorbance/time
Discussion 30%
This should be extensive consideration and interpretation of the results. Must include significance and application of the results in wider context. Should not just be a description of the findings (which should be in the results). Results should be discussed with respect to relevant scientific resources (books/journal articles). Should not simply be a restating of the literature given in the introduction, but rather using the literature to explain (and possibly extend) specific aspects of the results. Should be reflective – consideration of the experimental design with suggestions for improvement for follows up experiments.
Presentation and referencing: ~ 10 %
Clear and concise scientific English with high standard of spelling and grammar. Correct referencing style
Example type sources to use: chitturi S 2013 studies on protein content protease activity anti oxidase potential and web sites like; United States department of agriculture.
THERE ARE SOME ATTACHMENTS THAT COMES WITH THIS WORK THAT IS ESSENTIAL AS GUILDLINE FOR THE WRITING.
PLEASE ensure you understand the nature of the laboratory research paper that is to be written. Please use the attached matterial i have provided fo guldlines to how the paper should be written and also using research sources for background knowlege of work and to support your work. Take into concideration the tables and Figures(graphs) that need to be made to represent results. furthermore, please take into consideration the results data and calculations that needs to provided in this work. i put trust in you that you will reflect this piece of work well. Any details or questions please feel free to contact me to ask. Many Thanks Kind Regards
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