Team:ETH Zurich/Experiments 4
From 2013.igem.org
What about the hydolases ? How do they work and where do they come from ? Why do we use hydrolases ?
These enzymes have features that make them attractive as reporters. They are known to be relatively stable, exhibit activity under different conditions (e.g. pH and temperature), and catalyze the cleavage of various colorimetric and fluorescent substrates, which is ideal for visual screening (Kiernan, 2007).
Contents |
LacZ : beta-galactosidase
PhoA : Alkaline phosphatase
Aes : Carboxyl esterase
Nagz : Glycoside hydrolase
GusA : β-glucuronidase
The E. coli β-glucuronidase (GusA) is a tetrameric enzyme that hydrolyzes a wide variety of β-glucuronides (e.g. 6-chloro-3-indolyl-beta-D-glucuronide-cycloheylammonium salt). β-glucuronidase has no cofactors, nor any ionic requirements. The enzyme can tolerate a wide range of pH, with an optimum between 5.0 and 7.8, and it is reasonably resistant to thermal inactivation with a half-life at 55 about two hours (Jefferson, 1995).