Sandbox323: Difference between revisions
From Proteopedia
Jump to navigationJump to search
No edit summary |
No edit summary |
||
| Line 98: | Line 98: | ||
A linear equation of y=9.275x10^-7x+1x10^-4 with a coefficient of determination of 0.8492 was calculated using linear regression. | A linear equation of y=9.275x10^-7x+1x10^-4 with a coefficient of determination of 0.8492 was calculated using linear regression. | ||
[[Image:Tablebeerlaw.png| | [[Image:Tablebeerlaw.png|600px|left|thumb|Beer-Lambert Law sample calculation]] | ||
[[Image:Concentrationcoldmolarabsorb12000ph7.png|500px|left|thumb|]] | [[Image:Concentrationcoldmolarabsorb12000ph7.png|500px|left|thumb|]] | ||
| Line 104: | Line 104: | ||
Absorbance values can be transformed to units of concentration via the Beer-Lambert law. We must accept the approximation of the Molar Extinction Coefficient for PNPA hydrolysis at 428.7nm as 12000 M^-1 cm^-1. An example calculation is supplied in the table. Graphing time versus concentration and determining the slope of the line yields the enzyme's velocity in M/min. 1mg/mL of PNPA is saturating conditions which implies the Vmax is also the slope. The reaction volume total times Vmax yields Units of Enzyme Activity. This value can be used as a relative comparison tool for enzyme performance in given conditions. | Absorbance values can be transformed to units of concentration via the Beer-Lambert law. We must accept the approximation of the Molar Extinction Coefficient for PNPA hydrolysis at 428.7nm as 12000 M^-1 cm^-1. An example calculation is supplied in the table. Graphing time versus concentration and determining the slope of the line yields the enzyme's velocity in M/min. 1mg/mL of PNPA is saturating conditions which implies the Vmax is also the slope. The reaction volume total times Vmax yields Units of Enzyme Activity. This value can be used as a relative comparison tool for enzyme performance in given conditions. | ||
[[Image:Iceenzymeefficiencyscinotation.png| | [[Image:Iceenzymeefficiencyscinotation.png|600px|left|thumb|Enzyme Activity at neutral pH at cold temperature with Chloride cofactor]] | ||
The cold 4 degrees C reaction was changed to room temperature 20 degrees C. All other conditions remained constant to evaluate the effect of temperature on enzyme. Temperature increase negatively affects enzyme performance. Considering the cold loving nature of Chitinophaga pinensis, the enzyme being more active at a lower temperature is a reasonable conclusion. | The cold 4 degrees C reaction was changed to room temperature 20 degrees C. All other conditions remained constant to evaluate the effect of temperature on enzyme. Temperature increase negatively affects enzyme performance. Considering the cold loving nature of Chitinophaga pinensis, the enzyme being more active at a lower temperature is a reasonable conclusion. | ||
[[Image:Roomtempconcentrationtime.png|600px|left|thumb|Enzyme Activity at neutral pH at room temperature with Chloride cofactor]] | |||
[[Image:Sigfigroomtempenzymeactivity.png|600px|left|thumb|Enzyme Activity at neutral pH at room temperature with Chloride cofactor]] | |||
[[Image:Sigfigroomtempenzymeactivity.png| | |||
[[Image:Enzyme units percentage increase.png| | [[Image:Enzyme units percentage increase.png|600px|left|thumb|4 degrees C yields 42.2% increase in Units of Enzyme Activity цmol/minute]] | ||
===Discussion and Conclusion=== | ===Discussion and Conclusion=== | ||
