Difference between revisions of "Part:BBa K398029"
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This part was characterized at 37º C and pH=9.5, using recombinant strains carrying this part on the plasmid pSB1A2 by TU Delft 2010 iGEM team.  
 
This part was characterized at 37º C and pH=9.5, using recombinant strains carrying this part on the plasmid pSB1A2 by TU Delft 2010 iGEM team.  
  
 +
===Results===
 
[http://2010.igem.org/Team:TU_Delft#page=Project/alkane-degradation/results TU Delft 2010 results] suggest that the cell extracts obtained from the recombinant strain ''E. coli'' 029A, which expresses this part, have a dodecanal dehydrogenase activity twice as high as the control strain (''E. coli'' with the empty plasmid pSB1A2). This activity is equivalent to 33.98% of the cell extract of ''Pseudomonas putida'' growing on octane as sole carbon source.
 
[http://2010.igem.org/Team:TU_Delft#page=Project/alkane-degradation/results TU Delft 2010 results] suggest that the cell extracts obtained from the recombinant strain ''E. coli'' 029A, which expresses this part, have a dodecanal dehydrogenase activity twice as high as the control strain (''E. coli'' with the empty plasmid pSB1A2). This activity is equivalent to 33.98% of the cell extract of ''Pseudomonas putida'' growing on octane as sole carbon source.
  
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[[Image:TUDelftALDH_final.jpg|600px|thumb|center|Comparison of ALDH activities in the different strains tested in this study]]
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[[Image:TUDelftALDH_final.jpg|500px|thumb|left|Comparison of ALDH activities in the different strains tested in this study]]
  
  

Revision as of 16:56, 27 October 2010

ALDH generator (low expression)

Figure 1:Complete Alkane degradation pathway, ALDH is the 3rd step herein

ALDH is an aldehyde deyhydrogenase that facilitates the third step in alkane degradation, from n-alkanals to n-alkanoic acids (see figure 1), which can then be further degraded through β-oxidation

The [http://2010.igem.org/Team:TU_Delft#page=Project/alkane-degradation characterization of BBa_K398029] has been described on the TU Delft iGEM Team 2010 wiki.

Usage and Biology

Aldehyde dehydrogenase from the thermophile Geobacillus thermoleovorans B23. It functions as an octamer, requiring NAD+ as coenzyme. The optimum condition for activity lies at temperatures between 50 and 55 degrees celsius and and a pH of 10. You can see our original plasmid map below.

Comparison of ALDH activities in the different cell extracts tested in our study.

Characterization

This part was characterized at 37º C and pH=9.5, using recombinant strains carrying this part on the plasmid pSB1A2 by TU Delft 2010 iGEM team.

Results

[http://2010.igem.org/Team:TU_Delft#page=Project/alkane-degradation/results TU Delft 2010 results] suggest that the cell extracts obtained from the recombinant strain E. coli 029A, which expresses this part, have a dodecanal dehydrogenase activity twice as high as the control strain (E. coli with the empty plasmid pSB1A2). This activity is equivalent to 33.98% of the cell extract of Pseudomonas putida growing on octane as sole carbon source.

Comparison of ALDH activities in the different strains tested in this study



Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 37
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 37
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 37
  • 1000
    COMPATIBLE WITH RFC[1000]


[1] Kato, T., et al., Gene cloning and characterization of an aldehyde dehydrogenase from long-chain alkane-degrading Geobacillus thermoleovorans B23. Extremophiles, 2010. 14(1): p. 33-39.