Team:Cambridge/ProjectBioluminescence/Luciferase/WikiGeneticsLuxCDABE

From 2010.igem.org

(Difference between revisions)
(Luciferase)
(The Genetics of the Vibrio Lux operon - a short introduction)
Line 2: Line 2:
(Meighen, 1991)
(Meighen, 1991)
-
[[http://www.ncbi.nlm.nih.gov/pmc/articles/PMC372803/?tool=pubmed|Molecular biology of bacterial bioluminescence, Meighen, 1991]] - a very complete article on the nature of bioluminescence in this system.
+
[http://www.ncbi.nlm.nih.gov/pmc/articles/PMC372803/?tool=pubmed|Molecular biology of bacterial bioluminescence, Meighen, 1991] - a very complete article on the nature of bioluminescence in this system.
==Operon Structure==
==Operon Structure==
Line 18: Line 18:
*lux D codes for acyl-transferase
*lux D codes for acyl-transferase
*lux E makes the proteins needed for the enzyme acyl-protein synthetase.
*lux E makes the proteins needed for the enzyme acyl-protein synthetase.
 +
 +
Although the lux operon encodes the enzymes necessary for the bacteria to glow, bioluminescence is regulated by autoinduction and quorum sensing (the luxRI genes)
==Luciferase==
==Luciferase==
Line 34: Line 36:
  Reaction:  
  Reaction:  
-
  R-COOH + ATP + NADPH ------> R-CHO + AMP + PPi + NADP+      (Winfrey et al., 1997).  
+
  R-COOH + ATP + NADPH ------> R-CHO + AMP + PPi + NADP+      (Winfrey et al, 1997).  
-
Although the lux operon encodes the enzymes necessary for the bacteria to glow, bioluminescence is regulated by autoinduction and quorum sensing.
+
==LuxR and LuxI==

Revision as of 15:38, 19 July 2010

Contents

The Genetics of the Vibrio Lux operon - a short introduction

(Meighen, 1991) biology of bacterial bioluminescence, Meighen, 1991 - a very complete article on the nature of bioluminescence in this system.

Operon Structure

  • LuxCDABE => Emission system
  • LuxRI => Regulation system (several external and intrinsic factors appear to induce and inhibit the transcription of this gene set)


The lux operon has a known gene sequence of luxCDAB(F)E:

lux A and lux B code for the components of luciferase

lux CDE codes for a fatty acid reductase complex that makes the fatty acids necessary for the luciferase mechanism:

  • Lux C codes for the enzyme acyl-reductase
  • lux D codes for acyl-transferase
  • lux E makes the proteins needed for the enzyme acyl-protein synthetase.

Although the lux operon encodes the enzymes necessary for the bacteria to glow, bioluminescence is regulated by autoinduction and quorum sensing (the luxRI genes)

Luciferase

Luciferase produces blue/green light through the oxidation of reduced flavin mononucleotide (a standard biomolecule) and a long chain aldehyde by diatomic oxygen.

Reaction: 
FMNH2 + O2 + R-CHO ------> FMN + R-COOH + H2O + Light       (Silverman et al, 1984)

In order to generate the aldehyde needed in the reaction above, three additional enzymes are needed. The fatty acids needed for the reaction are pulled out from the fatty acid biosynthesis pathway by the enzyme acyl-transferase.

Acyl-trasferase reacts with acyl-ACP (from Fatty Acid pathway) to release R-COOH, a free fatty acid.

R-COOH is reduced by a two-enzyme system to an adehyde

Reaction: 
R-COOH + ATP + NADPH ------> R-CHO + AMP + PPi + NADP+       (Winfrey et al, 1997). 

LuxR and LuxI