Team:Bielefeld-Germany/Results/trametis
From 2012.igem.org
(→TVEL0 Activity Tests) |
|||
Line 12: | Line 12: | ||
<p class="more"> | <p class="more"> | ||
</html> | </html> | ||
- | + | [http://www.sigmaaldrich.com/catalog/product/sigma/51639?lang=de®ion=DE TVEL0] was characterized in terms of its activity to establish activity test protocols and to create a standard which can be used as a reference. | |
Line 21: | Line 21: | ||
==TVEL0 Activity Tests== | ==TVEL0 Activity Tests== | ||
- | |||
- | |||
===Initial Activity Test=== | ===Initial Activity Test=== | ||
[[File:Bielefeld2012_TVEL0_initial.jpg|thumbnail|450px|left|'''Figure 1:''']] | [[File:Bielefeld2012_TVEL0_initial.jpg|thumbnail|450px|left|'''Figure 1:''']] | ||
+ | To standardize activity test methods used for this project a [http://www.sigmaaldrich.com/catalog/product/sigma/51639?lang=de®ion=DE laccase from ''Trametes versicolor''] (TVEL0) was used. The optimal composition for activity measurements contains 140 µL of 0.03 mg mL<sup>-1</sup> concentrated TVEL0 laccase solution, 100 mM sodium acetate buffer, 0.1 mM ABTS, ad 200 µL H<sub>2</sub>O. With this approach activity in oxidizing ABTS of TVEL0 was measured over a time period of 5 minutes at 25°C. The saturation of the reaction was reached after 3 minutes when ~80% ABTS got oxidized (see figure 1). This result proofed the activity measurement method and can therefor be used as a positive control. | ||
+ | |||
+ | ===Optimal pH of TVEL0=== | ||
+ | |||
+ | |||
<html> | <html> |
Revision as of 18:56, 26 September 2012
Summary
[http://www.sigmaaldrich.com/catalog/product/sigma/51639?lang=de®ion=DE TVEL0] was characterized in terms of its activity to establish activity test protocols and to create a standard which can be used as a reference.
Contents |
TVEL0 Activity Tests
Initial Activity Test
To standardize activity test methods used for this project a [http://www.sigmaaldrich.com/catalog/product/sigma/51639?lang=de®ion=DE laccase from Trametes versicolor] (TVEL0) was used. The optimal composition for activity measurements contains 140 µL of 0.03 mg mL-1 concentrated TVEL0 laccase solution, 100 mM sodium acetate buffer, 0.1 mM ABTS, ad 200 µL H2O. With this approach activity in oxidizing ABTS of TVEL0 was measured over a time period of 5 minutes at 25°C. The saturation of the reaction was reached after 3 minutes when ~80% ABTS got oxidized (see figure 1). This result proofed the activity measurement method and can therefor be used as a positive control.
Optimal pH of TVEL0
55px |