Difference between revisions of "Part:BBa K4083004"

Revision as of 08:33, 20 October 2021

nadE gene of Pseudomonas aeruginosa

nadE from Pseudomonas aeruginosa is coding for NAD synthetase which synthesizes NAD+

Usage and Biology

Our team extracted nadE gene from P. aeruginosa to add it into novel plasmid called pRGPDuo2 for P. putida. Apart from nadE gene, planned to add rhlA and rhlB genes that are responsible for rhamnolipid synthesis. Thus, we predicted that dual expression of NAD synthetase and Rhamnosyltrnasferse can allow P. putida to express rhamnolipids in higher rates.

Pseudaminas aeuriginosa - is gram negative bacilus and opportunistic pathogen. NH(3) dependent NAD-synthetase converts deamido-NAD+ to NAD+ by ATP-dependent amidation [1]. ATP + deamido-NAD+ + NH4+ -> AMP + diphosphate + H+ + NAD+

The NAD+ is important for metabolism in organisms. NAD+ can reduce into NADH during cell digestion like glucolysis or Krebs cycle. Thus, more available NAD+ can lead to faster substrate catabolism. Moreover, NADH interacts with the electron transport chain where it releases one electron and one proton. As an electron moves, more protons exit the bacterial membrane which increases the proton gradient. Finally, to reach equilibrium, protons enter the cell by ATP synthase, and one proton can generate up to 3 ATP molecules this way. Thus, one NADH that releases one proton can generate 3 ATP molecules. Finally, NAD+ can interact with NAD kinase to convert into NADP+ which plays a direct role in biosynthesis of rhamnolipids. (Alberts et al., 2015)

Reference:

[1] UniProt. (n.d.). nadE - NH(3)-dependent NAD(+) synthetase - Pseudomonas aeruginosa (strain PA7) - nadE gene & protein. https://www.uniprot.org/uniprot/A6VD32 Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal XhoI site found at 208
Illegal XhoI site found at 475
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 400
Illegal NgoMIV site found at 470
Illegal NgoMIV site found at 691
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K4083004 short</partinfo>
-nadE from Pseudomonas aeruginosa is coding for NAD synthetase which synthesizes NAD+ from deamido-NAD+ by ATP-dependent amidation [1].
+nadE from Pseudomonas aeruginosa is coding for NAD synthetase which synthesizes NAD+
-ATP + deamido-NAD+ + NH4+ -> AMP + diphosphate + H+ + NAD+
-https://static.igem.org/mediawiki/parts/thumb/4/45/BBa_K4083004-NADS_reaction.png/799px-BBa_K4083004-NADS_reaction.png
+===Usage and Biology===
-Reference:
+Our team extracted nadE gene from P. aeruginosa to add it into novel plasmid called pRGPDuo2 for P. putida. Apart from nadE gene, planned to add rhlA and rhlB genes that are responsible for rhamnolipid synthesis. Thus, we predicted that dual expression of NAD synthetase and Rhamnosyltrnasferse can allow P. putida to express rhamnolipids in higher rates.
-[1] UniProt. (n.d.). nadE - NH(3)-dependent NAD(+) synthetase - Pseudomonas aeruginosa (strain PA7) - nadE gene & protein. https://www.uniprot.org/uniprot/A6VD32
 Pseudaminas aeuriginosa - is gram negative bacilus and opportunistic pathogen.
 NH(3) dependent NAD-synthetase converts deamido-NAD+ to NAD+ by ATP-dependent amidation [1].
 ATP + deamido-NAD+ + NH4+ -> AMP + diphosphate + H+ + NAD+
-The ammnoia is obtained by
 https://static.igem.org/mediawiki/parts/thumb/4/45/BBa_K4083004-NADS_reaction.png/799px-BBa_K4083004-NADS_reaction.png
+The NAD+ is important for metabolism in organisms. NAD+ can reduce into NADH during cell digestion like glucolysis or Krebs cycle. Thus, more available NAD+ can lead to faster substrate catabolism.
+Moreover, NADH interacts with the electron transport chain where it releases one electron and one proton. As an electron moves, more protons exit the bacterial membrane which increases the proton gradient. Finally, to reach equilibrium, protons enter the cell by ATP synthase, and one proton can generate up to 3 ATP molecules this way. Thus, one NADH that releases one proton can generate 3 ATP molecules. Finally, NAD+ can interact with NAD kinase to convert into NADP+ which plays a direct role in biosynthesis of rhamnolipids. (Alberts et al., 2015)
-Our team extracted nadE gene from P. aeruginosa to add it into novel plasmid called pRGPDuo2 for P. putida. Apart from nadE gene, planned to add rhlA and rhlB genes that are responsible for rhamnolipid synthesis. Thus, we predicted that dual expression of NAD synthetase and Rhamnosyltrnasferse can allow P. putida to express rhamnolipids in higher rates.
+Reference:
-The NAD+ is important for metabolism in organisms. NAD+ can reduce into NADH during cell digestion like glucolysis or Krebs cycle. Thus, more available NAD+ can lead to faster substrate catabolism.
+[1] UniProt. (n.d.). nadE - NH(3)-dependent NAD(+) synthetase - Pseudomonas aeruginosa (strain PA7) - nadE gene & protein. https://www.uniprot.org/uniprot/A6VD32
-Moreover, NADH interacts with the electron transport chain where it releases one electron and one proton. As an electron moves, more protons exit the bacterial membrane which increases the proton gradient. Finally, to reach equilibrium, protons enter the cell by ATP synthase, and one proton can generate up to 3 ATP molecules this way. Thus, one NADH that releases one proton can generate 3 ATP molecules. Finally, NAD+ can interact with NAD kinase to convert into NADP+ which plays a direct role in biosynthesis of rhamnolipids. (Alberts et al., 2015) <!-- Add more about the biology of this part here
+<!-- Add more about the biology of this part here
 ===Usage and Biology===