Dan Tawfik lab: Directed evolution: Difference between revisions
Jump to navigation
Jump to search
No edit summary |
m Directed evolution moved to Dan Tawfik lab: Directed evolution: To honor Dan Tawfik and make the title fit better to the content |
||
| (2 intermediate revisions by one other user not shown) | |||
| Line 1: | Line 1: | ||
<StructureSection load='Crdes1.pdb' size=' | <StructureSection load='Crdes1.pdb' size='450' frame='true' side='right' scene='3iio/Int/1' > | ||
== I) Kemp eliminase == | == I) Kemp eliminase == | ||
A series of [http://en.wikipedia.org/wiki/Protein_design computationally designed enzymes] that catalyze the Kemp elimination have described. Kemp eliminase (KE07) has <scene name='3iio/Int/10'>TIM barrel scaffold</scene>. The Kemp elimination of <scene name='3iio/Int/4'>5-nitrobenzisoxazole</scene> was chosen as a model reaction for [http://en.wikipedia.org/wiki/Proton#Hydrogen_as_proton proton (H)] transfer from [http://en.wikipedia.org/wiki/Carbon carbon], simultaneously with the cut of the [http://en.wikipedia.org/wiki/Nitrogen nitrogen]–[http://en.wikipedia.org/wiki/Oxygen oxygen] (N-O) bond, resulting in <scene name='3iio/Int/8'>cyanophenol product</scene>. Such reaction is a critical step in many [http://en.wikipedia.org/wiki/Enzyme enzymatic reactions]. The [http://en.wikipedia.org/wiki/Enzyme_catalysis catalytic] base (E101), the general acid/[http://en.wikipedia.org/wiki/Hydrogen_bond H-bond] donor (K222), and the stacking residue (W50) make interactions with the 5-nitrobenzisoxazole at the <scene name='3iio/Int/9'>active site of KE07</scene>. [http://en.wikipedia.org/wiki/Directed_evolution Directed evolution] can significantly improve the stability, expression and activity of enzymes. In the catalytically improved directed evolutionary variants of KE07 containing the <scene name='3iio/Ali/7'>Ile7Asp mutation</scene>, Asp7 breaks the Glu101–Lys222 salt bridge (for example [[3iiv]], chain A is shown). | A series of [http://en.wikipedia.org/wiki/Protein_design computationally designed enzymes] that catalyze the Kemp elimination have described. Kemp eliminase (KE07) has <scene name='3iio/Int/10'>TIM barrel scaffold</scene>. The Kemp elimination of <scene name='3iio/Int/4'>5-nitrobenzisoxazole</scene> was chosen as a model reaction for [http://en.wikipedia.org/wiki/Proton#Hydrogen_as_proton proton (H)] transfer from [http://en.wikipedia.org/wiki/Carbon carbon], simultaneously with the cut of the [http://en.wikipedia.org/wiki/Nitrogen nitrogen]–[http://en.wikipedia.org/wiki/Oxygen oxygen] (N-O) bond, resulting in <scene name='3iio/Int/8'>cyanophenol product</scene>. Such reaction is a critical step in many [http://en.wikipedia.org/wiki/Enzyme enzymatic reactions]. The [http://en.wikipedia.org/wiki/Enzyme_catalysis catalytic] base (E101), the general acid/[http://en.wikipedia.org/wiki/Hydrogen_bond H-bond] donor (K222), and the stacking residue (W50) make interactions with the 5-nitrobenzisoxazole at the <scene name='3iio/Int/9'>active site of KE07</scene>. [http://en.wikipedia.org/wiki/Directed_evolution Directed evolution] can significantly improve the stability, expression and activity of enzymes. In the catalytically improved directed evolutionary variants of KE07 containing the <scene name='3iio/Ali/7'>Ile7Asp mutation</scene>, Asp7 breaks the Glu101–Lys222 salt bridge (for example [[3iiv]], chain A is shown). | ||