Team:ETH Zurich/Experiments 3
From 2013.igem.org
Line 35: | Line 35: | ||
<td>Magenta glucuronide</td> | <td>Magenta glucuronide</td> | ||
<td>6-chloro-3-indolyl-beta-D-glucuronide-cycloheylammonium salt</td> | <td>6-chloro-3-indolyl-beta-D-glucuronide-cycloheylammonium salt</td> | ||
- | <td style="color: | + | <td style="color:Salmon (pinkish orange)">Red[[Image:rot.png|600px|left|thumb|]]</td> |
</tr> | </tr> | ||
<tr> | <tr> |
Revision as of 21:15, 28 September 2013
Contents |
Enzyme-substrate reactions
The enzyme substrate reactions take less than 5 minutes and are visible by eye.
Our minesweeper is becoming better and better so keep on track for updates !
In Figure 1 you can see the different hydrolases we use.
Hydrolase | Complementary substrate / IUPAC name | Visible color | Concentration[M] | Time(s)for colorimetric response | ||
---|---|---|---|---|---|---|
LacZ | Beta-Galactosidase | X-Gal | 5-Bromo-4chloro-3-indolyl-beta-galactopyranoside | Blue | ||
LacZ | Beta-Galactosidase | Green-beta-D-Gal | N-Methyl-3-indolyl-beta-D_galactopyranoside | Green | ||
GusA | Beta-glucuronidase | Magenta glucuronide | 6-chloro-3-indolyl-beta-D-glucuronide-cycloheylammonium salt | Red | ||
PhoA | Alkaline phosphatase | pNPP | 4-Nitrophenylphosphatedi(tris) salt | Yellow | ||
Aes | Acetyl esterase | Magenta butyrate | 5-bromo-6-chloro-3-indoxyl butyrate | Magenta | ||
NagZ | Glycoside hydrolase | X-glucosaminide X-Glucnac | 5-bromo-4-chloro-3-indolyl-N-acetyl-beta-D-glucosaminide | Blue |
Beta-galactosidase(LacZ)
Chemical conversion
Colorimetric response in liquid culture
Colorimetric response on LB-Agar
Alkaline phosphatase(PhoA)
Chemical conversion
Colorimetric response in liquid culture
Colorimetric response on LB-Agar
Acetyl esterase(Aes)
Chemical conversion
Colorimetric response in liquid culture
Colorimetric response on LB-Agar
Beta-glucuronidase(GusA)
Chemical conversion
Colorimetric response in liquid culture
Colorimetric response on LB-Agar
β-N-Acetylglucosaminidase(NagZ)
Chemical conversion
Colorimetric response in liquid culture
Colorimetric response on LB-Agar
Do the substrates and enzymes cross-react ?
An enzyme-substrate test matrix (Figure 6) was established to test each substrate against each enzyme. The results were as expected (Figure 6.2) and no cross reaction is visible. The NagZ-X - glucosaminide X-Glunac reveal some difficulties in the liquid culture as well as on the agar plate.