Difference between revisions of "Part:BBa K398029"
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__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K398029 short</partinfo> | <partinfo>BBa_K398029 short</partinfo> | ||
| − | + | [[Image:TUDelft_Alkane_degradation_route.png|right|300px|thumb|'''Figure 1:'''Complete Alkane degradation pathway, ALDH is the 3rd step herein]] | |
| − | ALDH | + | 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. | 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. | ||
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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. | 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. | ||
| − | [[Image:TUDelftALDH_map.jpg| | + | [[Image:TUDelftALDH_map.jpg|400px|thumb|right|Comparison of ALDH activities in the different cell extracts tested in our study.]] |
===Characterization=== | ===Characterization=== | ||
Revision as of 16:54, 27 October 2010
ALDH generator (low expression)
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.
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.
[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.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 37
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 37
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 37
- 1000COMPATIBLE 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.