Coding

Part:BBa_K857000

Designed by: yale yuen   Group: iGEM12_UCMerced_Bobcats   (2012-10-02)


acetaldehyde dehydrogenase

Coding protein for acetaldehyde dehydrogenase convert acetaldehyde into acetyl-CoA.

CH3CHO + NAD+ + CoA → acetyl-CoA + NADH + H+

This part was designed by the 2012 iGEM UC Merced team and it is a gene that is involved in the E. Coli glucose metabolism pathway. Our team characterized this part by performing an in silico knockout of this gene. Using quantitative analysis, we discovered that its deletion does not affect the growth of E. Coli when under optimal conditions to produce a 4:1 hydrogen production to glucose uptake ratio. More importantly, we identified possible non-lethal gene deletions that may help maximize hydrogen production in the dark fermentation process.

First, we measured the growth of E. Coli bacteria under the conditions in which the insertion of mhpF allows the production of hydrogen to glucose uptake ratio to be 4:1. This was done using the Constraint Model Based Reconstruction and Analysis (COBRA) toolbox on Matlab platform which allowed us to constrain the uptake of glucose and hydrogen release to a ratio of 4:1 on our E. Coli glucose metabolism model. Our results were performed using Flux Balance Analysis (FBA) on the Core E. Coli Model by the Systems Biology Research Group at the University of California generated from the current gene expression data on E. coli. T--NYU_Abu_Dhabi--characterization-graph1.png

Figure 1. Demonstrates the growth of E.coli when subject to the hydrogen release to glucose uptake requirement.

Our results showed that when the MhpF gene is introduced, E. Coli growth is higher under anaerobic conditions than aerobic conditions when under stress requirement to produce a 4:1 hydrogen to glucose ratio. This was achieved by setting oxygen flux values to both unlimited and zero for aerobic and anaerobic conditions respectively, whilst measuring E. coli’s growth in rate per hour. Growth measurements were done as a quantitative measure of how E.coli performs against the gene modification added to its metabolism.

Furthermore, our analysis from Flux Balance Analysis (FBA) validated team UC Merced hypothesis that removing IdhA, pfIB and adhE from E. coli fermentation pathway and replacing it with mhpF, pyruvate decarboxylase and ferredoxin oxidoreductase. This was achieved by performing in silico knockouts of IdhA, pfIB and adhE on the E. coli model and multiple gene insertions of mhpF, pyruvate decarboxylate and ferredoxin and looking at how the shift in metabolism network improve hydrogen yield under anaerobic condition.

1190px-T--NYU_Abu_Dhabi--character-graph2.png T--NYU_Abu_Dhabi--character-graph3.png

Figure 2.Shift in metabolism flux network when E. coli model is subjected to hydrogen release to glucose uptake 4:1 ratio under aerobic conditions (left) and anaerobic conditions(right).

Next, we analyzed the effect of deleting the mhpF gene on the growth rate of E. Coli (in biomass/hour). The quantitative analysis shows that the growth remained the same, which indicates that the mhpF gene does not affect the growth rate of E. Coli. This served as motivation to perform more single gene deletions on the E.coli model. We analyzed the effect of creating knockouts of all the genes involved in glucose metabolism on the bacteria by examining the lethality of deleting each gene. Team UC Merced attempted to create knockouts of three genes, and below we present suggestions regarding the genes in the dark fermentation process that can be deleted without being lethal. Deletions of genes in the dark fermentation process, a form of anaerobic conversion would facilitate the conversion of organic substrate to biohydrogen. (see Table 1) For instance our analysis from the gene deletion suggested that the deletion zwf gene( glucose-6-phosphate 1-dehydrogenase) which is principal in the pentose pathway would be lethal to the E. coli. It turns out that zwf and mphF gene is overexpressed in the dark fermentation process. Even though both are very necessary for hydrogen production, mphF deletion would not result in much difference in the flux. Further our metabolic network flux predicts that the overexpression of zwf leads to the diversion of carbon flux through the PP pathway and hence result in an increase in the efficiency of the production of H2. Finally, a robustness analysis was performed to verify the validity of the results by setting fixed amounts of glucose and hydrogen and measuring the growth as a function of the oxygen concentration. This analysis was performed to demonstrate the strength of our model's prediction. The linear graph predicts a linear rise in growth with an intermittent increase in glucose intake. This agrees and supports our previous data which suggested that mphF is implied in the increase in growth of E.coli when subject to the condition stipulated above.

T--NYU_Abu_Dhabi--character-graph4.jpeg

In sum , using metabolic modeling allow for in silico gene knockouts and metabolic analysis that unveiled non lethal gene deletions and insertion that would prove useful for the generation of biohydrogen from E.coli. Using metabolic modeling, it is easier to generate genetic changes that would optimize hydrogen production from E.coli. graph robustness


Finally, a robustness analysis was performed to verify the validity of the results by setting fixed amounts of glucose and hydrogen and measuring the growth as a function of the oxygen concentration. Table 1: Possible gene deletions and their measure effects on E. coli.

 

<tbody> </tbody>

LOCUS

GENE

PRODUCT

FUNCTION

NOTE

LETHALITY

b0008

talB

transaldolase B

enzyme; Central intermediary metabolism:Non-oxidative branch, pentose pathway

 

not lethal

b0118

acnB

aconitate hydratase 2; aconitase B;2-methyl-cis-aconitate hydratase

enzyme; Energy metabolism, carbon: TCA cycle

aconitate hydrase B

not lethal

b0351

mhpF

acetaldehyde-CoA dehydrogenase II, NAD-binding

enzyme; Degradation of small molecules: Carboncompounds

acetaldehyde dehydrogenase

not lethal

b0356

frmA

alcohol dehydrogenase class III;glutathione-dependent formaldehyde dehydrogenase

enzyme; Energy metabolism, carbon:Fermentation

alcohol dehydrogenase class III; formaldehydedehydrogenase, glutathione-dependent

not lethal

b0451

amtB

ammonium transporter

putative transport; Central intermediarymetabolism: Pool, multipurpose conversions

probable ammonium transporter

not lethal

b0474

adk

adenylate kinase

enzyme; Purine ribonucleotide biosynthesis

adenylate kinase activity; pleiotropic effects onglycerol-3-phosphate acyltransferase activity

not lethal

b0485

glsA

glutaminase 1

enzyme; l-glutamine catabolism

putative glutaminase

not lethal

b0733

cydA

cytochrome d terminal oxidase, subunit I

enzyme; Energy metabolism, carbon: Electrontransport

cytochrome d terminal oxidase, polypeptide subunitI

not lethal

b0734

cydB

cytochrome d terminal oxidase, subunit II

enzyme; Energy metabolism, carbon: Electrontransport

cytochrome d terminal oxidase polypeptide subunitII

not lethal

b0755

gpmA

phosphoglyceromutase 1

enzyme; Energy metabolism, carbon: Glycolysis

 

not lethal

b0809

glnQ

glutamine transporter subunit

transport; Transport of small molecules: Aminoacids, amines

ATP-binding component of glutamine high-affinitytransport system

not lethal

b0810

glnP

glutamine transporter subunit

transport; Transport of small molecules: Aminoacids, amines

glutamine high-affinity transport system; membranecomponent

not lethal

b0811

glnH

glutamine transporter subunit

transport; Transport of small molecules: Aminoacids, amines

periplasmic glutamine-binding protein; permease

not lethal

b0875

aqpZ

aquaporin Z

transport; Transport of small molecules: Other

transmembrane water channel; aquaporin Z

not lethal

b0902

pflA

pyruvate formate-lyase 1-activating enzyme;[formate-C-acetyltransferase 1]-activating enzyme; PFLactivase

enzyme; Energy metabolism, carbon: Anaerobicrespiration

 

not lethal

b0903

pflB

formate C-acetyltransferase 1, anaerobic;pyruvate formate-lyase 1

enzyme; Energy metabolism, carbon: Anaerobicrespiration

formate acetyltransferase 1

not lethal

b0904

focA

formate channel

putative transport; Degradation of smallmolecules: Carbon compounds

probable formate transporter (formate channel 1)

not lethal

b0978

cbdA

cytochrome bd-II oxidase, subunit I

putative enzyme; Energy metabolism, carbon:Electron transport

 

not lethal

b0979

cbdB

cytochrome bd-II oxidase, subunit II

putative enzyme; Energy metabolism, carbon:Electron transport

probable third cytochrome oxidase, subunit II

not lethal

b1101

ptsG

fused glucose-specific PTS enzymes: IIBcomponent/IIC component

transport; Transport of small molecules:Carbohydrates, organic acids, alcohols

PTS system, glucose-specific IIBC component

not lethal

b1241

adhE

fused acetaldehyde-CoAdehydrogenase/iron-dependent alcoholdehydrogenase/pyruvate-formate lyase deactivase

enzyme; Energy metabolism, carbon:Fermentation

CoA-linked acetaldehyde dehydrogenase andiron-dependent alcohol dehydrogenase;pyruvate-formate-lyase deactivase

not lethal

b1276

acnA

aconitate hydratase 1; aconitase A

enzyme; Energy metabolism, carbon: TCA cycle

aconitate hydrase 1

not lethal

b1297

puuA

glutamate--putrescine ligase

enzyme; Degradation of small molecules: Carboncompounds

putative glutamine synthetase

not lethal

b1380

ldhA

fermentative D-lactate dehydrogenase,NAD-dependent

enzyme; Energy metabolism, carbon:Fermentation

 

not lethal

b1478

adhP

ethanol-active dehydrogenase/acetaldehyde-activereductase

enzyme; Energy metabolism, carbon: Anaerobicrespiration

alcohol dehydrogenase

not lethal

b1479

maeA

malate dehydrogenase, decarboxylating,NAD-requiring; malic enzyme

enzyme; Central intermediary metabolism:Gluconeogenesis

NAD-linked malate dehydrogenase (malic enzyme)

not lethal

b1524

glsB

glutaminase 2

enzyme; l-glutamine catabolism

putative glutaminase

not lethal

b1602

pntB

pyridine nucleotide transhydrogenase, betasubunit

enzyme; Central intermediary metabolism: Pool,multipurpose conversions

 

not lethal

b1603

pntA

pyridine nucleotide transhydrogenase, alphasubunit

enzyme; Central intermediary metabolism: Pool,multipurpose conversions

 

not lethal

b1611

fumC

fumarate hydratase (fumarase C),aerobic ClassII

enzyme; Energy metabolism, carbon: TCA cycle

fumarase C

not lethal

b1612

fumA

fumarate hydratase (fumarase A), aerobic ClassI

enzyme; Energy metabolism, carbon: TCA cycle

fumarase A isozyme

not lethal

b1621

malX

maltose and glucose-specific PTS enzyme IIBcomponent and IIC component

transport; Transport of small molecules:Carbohydrates, organic acids, alcohols

PTS system, maltose and glucose-specific IIABCcomponent

not lethal

b1676

pykF

pyruvate kinase I

enzyme; Energy metabolism, carbon: Glycolysis

pyruvate kinase I (formerly F), fructosestimulated

not lethal

b1702

ppsA

phosphoenolpyruvate synthase

enzyme; Central intermediary metabolism:Gluconeogenesis

 

not lethal

b1723

pfkB

6-phosphofructokinase II

enzyme; Energy metabolism, carbon: Glycolysis

6-phosphofructokinase II; suppressor of pfkA

not lethal

b1773

ydjI

putative aldolase

putative enzyme; Not classified

 

not lethal

b1812

pabB

aminodeoxychorismate synthase, subunit I

enzyme; Biosynthesis of cofactors, carriers:Folic acid

p-aminobenzoate synthetase, component I

not lethal

b1817

manX

fused mannose-specific PTS enzymes: IIAcomponent/IIB component

enzyme; Transport of small molecules:Carbohydrates, organic acids, alcohols

PTS enzyme IIAB, mannose-specific

not lethal

b1818

manY

mannose-specific enzyme IIC component of PTS

transport; Transport of small molecules:Carbohydrates, organic acids, alcohols

PTS enzyme IIC, mannose-specific

not lethal

b1819

manZ

mannose-specific enzyme IID component of PTS

transport; Transport of small molecules:Carbohydrates, organic acids, alcohols

PTS enzyme IID, mannose-specific

not lethal

b1849

purT

phosphoribosylglycinamide formyltransferase 2

enzyme; Purine ribonucleotide biosynthesis

 

not lethal

b1854

pykA

pyruvate kinase II

enzyme; Energy metabolism, carbon: Glycolysis

pyruvate kinase II, glucose stimulated

not lethal

b2097

fbaB

fructose-bisphosphate aldolase class I

   

not lethal

b2133

dld

D-lactate dehydrogenase, FAD-binding, NADHindependent

enzyme; Energy metabolism, carbon: Aerobicrespiration

D-lactate dehydrogenase, FAD protein, NADHindependent

not lethal

b2296

ackA

acetate kinase A and propionate kinase 2

enzyme; Energy metabolism, carbon: Electrontransport

acetate kinase

not lethal

b2297

pta

phosphate acetyltransferase

enzyme; Degradation of small molecules: Carboncompounds

phosphotransacetylase

not lethal

b2417

crr

glucose-specific enzyme IIA component of PTS

enzyme; Transport of small molecules:Carbohydrates, organic acids, alcohols

PTS system, glucose-specific IIA component

not lethal

b2458

eutD

phosphate acetyltransferase

putative enzyme; Degradation of smallmolecules: Amines

ethanolamine utilization; homolog of Salmonellaacetyl/butyryl P transferase

not lethal

b2463

maeB

malic enzyme: putativeoxidoreductase/phosphotransacetylase

putative enzyme; Not classified

putative multimodular enzyme

not lethal

b2464

talA

transaldolase A

enzyme; Central intermediary metabolism:Non-oxidative branch, pentose pathway

 

not lethal

b2465

tktB

transketolase 2, thiamine triphosphate-binding

enzyme; Central intermediary metabolism:Non-oxidative branch, pentose pathway

transketolase 2 isozyme

not lethal

b2492

focB

putative formate transporter

putative transport; Transport of smallmolecules: Carbohydrates, organic acids, alcohols

probable formate transporter (formate channel 2)

not lethal

b2579

grcA

autonomous glycyl radical cofactor

putative enzyme; Energy metabolism, carbon:Anaerobic respiration

putative formate acetyltransferase

not lethal

b2587

kgtP

alpha-ketoglutarate transporter

transport; Transport of small molecules:Carbohydrates, organic acids, alcohols

alpha-ketoglutarate permease

not lethal

b2914

rpiA

ribose 5-phosphate isomerase, constitutive

enzyme; Central intermediary metabolism:Non-oxidative branch, pentose pathway

ribosephosphate isomerase, constitutive

not lethal

b2925

fbaA

fructose-bisphosphate aldolase, class II

enzyme; Energy metabolism, carbon: Glycolysis

 

not lethal

b2935

tktA

transketolase 1, thiamine triphosphate-binding

enzyme; Central intermediary metabolism:Non-oxidative branch, pentose pathway

transketolase 1 isozyme

not lethal

b2975

glcA

glycolate transporter

putative transport; Not classified

putative permease

not lethal

b2976

glcB

malate synthase G

enzyme; Central intermediary metabolism:Glyoxylate bypass

 

not lethal

b2987

pitB

phosphate transporter

transport; Transport of small molecules:Anions

low-affinity phosphate transport

not lethal

b3114

tdcE

pyruvate formate-lyase 4/2-ketobutyrateformate-lyase

putative enzyme; Energy metabolism, carbon:Anaerobic respiration

probable formate acetyltransferase 3

not lethal

b3115

tdcD

propionate kinase/acetate kinase C, anaerobic

putative enzyme; Not classified

propionate kinase/acetate kinase II, anaerobic

not lethal

b3212

gltB

glutamate synthase, large subunit

enzyme; Central intermediary metabolism: Pool,multipurpose conversions

 

not lethal

b3213

gltD

glutamate synthase, 4Fe-4S protein, smallsubunit

enzyme; Central intermediary metabolism: Pool,multipurpose conversions

glutamate synthase, small subunit

not lethal

b3386

rpe

D-ribulose-5-phosphate 3-epimerase

enzyme; Central intermediary metabolism:Non-oxidative branch, pentose pathway

 

not lethal

b3403

pck

phosphoenolpyruvate carboxykinase [ATP]

enzyme; Central intermediary metabolism:Gluconeogenesis

 

not lethal

b3493

pitA

phosphate transporter, low-affinity; telluriteimporter

transport; Transport of small molecules:Anions

low-affinity phosphate transport

not lethal

b3528

dctA

C4-dicarboxylic acid, orotate and citratetransporter

transport; Transport of small molecules:Carbohydrates, organic acids, alcohols

uptake of C4-dicarboxylic acids

not lethal

b3603

lldP

L-lactate permease

transport; Transport of small molecules:Carbohydrates, organic acids, alcohols

 

not lethal

b3612

gpmM

phosphoglycero mutase III, cofactor-independent

putative enzyme; Not classified

putative 2,3-bisphosphoglycerate-independentphosphoglycerate mutase

not lethal

b3739

atpI

ATP synthase, membrane-bound accessory factor

enzyme; ATP-proton motive forceinterconversion

membrane-bound ATP synthase, dispensable protein,affects expression of atpB

not lethal

b3870

glnA

glutamine synthetase

enzyme; Amino acid biosynthesis: Glutamine

 

not lethal

b3916

pfkA

6-phosphofructokinase I

enzyme; Energy metabolism, carbon: Glycolysis

 

not lethal

b3925

glpX

fructose 1,6-bisphosphatase II

phenotype; Not classified

not required for growth on glycerol

not lethal

b3951

pflD

putative glycine radical domain-containingpyruvate formate-lyase

enzyme; Energy metabolism, carbon: Anaerobicrespiration

formate acetyltransferase 2

not lethal

b3952

pflC

putative [formate-C-acetyltransferase2]-activating enzyme; pyruvate formate-lyase 1-activatingenzyme

putative enzyme; Energy metabolism, carbon:Anaerobic respiration

probable pyruvate formate lyase activating enzyme2

not lethal

b3962

sthA

pyridine nucleotide transhydrogenase, soluble

putative enzyme; Not classified

 

not lethal

b4014

aceB

malate synthase A

enzyme; Central intermediary metabolism:Glyoxylate bypass

 

not lethal

b4015

aceA

isocitrate lyase

enzyme; Central intermediary metabolism:Glyoxylate bypass

 

not lethal

b4077

gltP

glutamate/aspartate:proton symporter

transport; Transport of small molecules: Aminoacids, amines

glutamate-aspartate symport protein

not lethal

b4090

rpiB

ribose 5-phosphate isomerase B/allose6-phosphate isomerase

enzyme; Central intermediary metabolism:Non-oxidative branch, pentose pathway

ribose 5-phosphate isomerase B

not lethal

b4122

fumB

anaerobic class I fumarate hydratase (fumaraseB)

enzyme; Energy metabolism, carbon: TCA cycle

fumarase Bisozyme

not lethal

b4151

frdD

fumarate reductase (anaerobic), membrane anchorsubunit

enzyme; Energy metabolism, carbon: Anaerobicrespiration

fumarate reductase, anaerobic, membrane anchorpolypeptide

not lethal

b4152

frdC

fumarate reductase (anaerobic), membrane anchorsubunit

enzyme; Energy metabolism, carbon: Anaerobicrespiration

fumarate reductase, anaerobic, membrane anchorpolypeptide

not lethal

b4153

frdB

fumarate reductase (anaerobic), Fe-S subunit

enzyme; Energy metabolism, carbon: Anaerobicrespiration

fumarate reductase, anaerobic, iron-sulfur proteinsubunit

not lethal

b4154

frdA

anaerobic fumarate reductase catalytic andNAD/flavoprotein subunit

enzyme; Energy metabolism, carbon: Anaerobicrespiration

fumarate reductase, anaerobic, flavoproteinsubunit

not lethal

b4232

fbp

fructose-1,6-bisphosphatase I

enzyme; Central intermediary metabolism:Gluconeogenesis

 

not lethal

b4301

sgcE

putative epimerase

putative enzyme; Not classified

 

not lethal

b4395

ytjC

phosphatase

enzyme; Not classified

phosphoglyceromutase 2

not lethal

b0114

aceE

pyruvate dehydrogenase, decarboxylase componentE1, thiamine triphosphate-binding

enzyme; Energy metabolism, carbon: Pyruvatedehydrogenase

pyruvate dehydrogenase (decarboxylase component)

lethal

b0115

aceF

pyruvate dehydrogenase,dihydrolipoyltransacetylase component E2

enzyme; Energy metabolism, carbon: Pyruvatedehydrogenase

pyruvate dehydrogenase (dihydrolipoyltransacetylasecomponent)

lethal

b0116

lpd

dihydrolipoyl dehydrogenase; E3 component ofpyruvate and 2-oxoglutarate dehydrogenases complexes;glycine cleavage system L protein; dihydrolipoamidedehydrogenase

enzyme; Energy metabolism, carbon: Pyruvatedehydrogenase

lipoamide dehydrogenase (NADH); component of2-oxodehydrogenase and pyruvate complexes; L-protein ofglycine cleavage complex

lethal

b0720

gltA

citrate synthase

enzyme; Energy metabolism, carbon: TCA cycle

 

lethal

b0721

sdhC

succinate dehydrogenase, membrane subunit, bindscytochrome b556

enzyme; Energy metabolism, carbon: TCA cycle

succinate dehydrogenase, cytochrome b556

lethal

b0722

sdhD

succinate dehydrogenase, membrane subunit, bindscytochrome b556

enzyme; Energy metabolism, carbon: TCA cycle

succinate dehydrogenase, hydrophobic subunit

lethal

b0723

sdhA

succinate dehydrogenase, flavoprotein subunit

enzyme; Energy metabolism, carbon: TCA cycle

 

lethal

b0724

sdhB

succinate dehydrogenase, FeS subunit

enzyme; Energy metabolism, carbon: TCA cycle

succinate dehydrogenase, iron sulfur protein

lethal

b0726

sucA

2-oxoglutarate decarboxylase, thiaminetriphosphate-binding

enzyme; Energy metabolism, carbon: TCA cycle

2-oxoglutarate dehydrogenase (decarboxylasecomponent)

lethal

b0727

sucB

dihydrolipoyltranssuccinase

enzyme; Energy metabolism, carbon: TCA cycle

2-oxoglutarate dehydrogenase(dihydrolipoyltranssuccinase E2 component)

lethal

b0728

sucC

succinyl-CoA synthetase, beta subunit

enzyme; Energy metabolism, carbon: TCA cycle

 

lethal

b0729

sucD

succinyl-CoA synthetase, NAD(P)-binding, alphasubunit

enzyme; Energy metabolism, carbon: TCA cycle

succinyl-CoA synthetase, alpha subunit

lethal

b0767

pgl

6-phosphogluconolactonase

orf; Not classified

putative isomerase

lethal

b1136

icd

isocitrate dehydrogenase; e14 prophageattachment site; tellurite reductase

enzyme; Energy metabolism, carbon: TCA cycle;Phage or Prophage Related

isocitrate dehydrogenase, specific for NADP+

lethal

b1761

gdhA

glutamate dehydrogenase, NADP-specific

enzyme; Amino acid biosynthesis: Glutamate

NADP-specific glutamate dehydrogenase

lethal

b1779

gapA

glyceraldehyde-3-phosphate dehydrogenase A

enzyme; Energy metabolism, carbon: Glycolysis

 

lethal

b1852

zwf

glucose-6-phosphate 1-dehydrogenase

enzyme; Energy metabolism, carbon: Oxidativebranch, pentose pathway

 

lethal

b2029

gnd

6-phosphogluconate dehydrogenase,decarboxylating

enzyme; Energy metabolism, carbon: Oxidativebranch, pentose pathway

 

lethal

b2276

nuoN

NADH:ubiquinone oxidoreductase, membrane subunitN

enzyme; Energy metabolism, carbon: Aerobicrespiration

NADH dehydrogenase I chain N

lethal

b2277

nuoM

NADH:ubiquinone oxidoreductase, membrane subunitM

enzyme; Energy metabolism, carbon: Aerobicrespiration

NADH dehydrogenase I chain M

lethal

b2278

nuoL

NADH:ubiquinone oxidoreductase, membrane subunitL

enzyme; Energy metabolism, carbon: Aerobicrespiration

NADH dehydrogenase I chain L

lethal

b2279

nuoK

NADH:ubiquinone oxidoreductase, membrane subunitK

enzyme; Energy metabolism, carbon: Aerobicrespiration

NADH dehydrogenase I chain K

lethal

b2280

nuoJ

NADH:ubiquinone oxidoreductase, membrane subunitJ

enzyme; Energy metabolism, carbon: Aerobicrespiration

NADH dehydrogenase I chain J

lethal

b2281

nuoI

NADH:ubiquinone oxidoreductase, chain I

enzyme; Energy metabolism, carbon: Aerobicrespiration

NADH dehydrogenase I chain I

lethal

b2282

nuoH

NADH:ubiquinone oxidoreductase, membrane subunitH

enzyme; Energy metabolism, carbon: Aerobicrespiration

NADH dehydrogenase I chain H

lethal

b2283

nuoG

NADH:ubiquinone oxidoreductase, chain G

enzyme; Energy metabolism, carbon: Aerobicrespiration

NADH dehydrogenase I chain G

lethal

b2284

nuoF

NADH:ubiquinone oxidoreductase, chain F

enzyme; Energy metabolism, carbon: Aerobicrespiration

NADH dehydrogenase I chain F

lethal

b2285

nuoE

NADH:ubiquinone oxidoreductase, chain E

enzyme; Energy metabolism, carbon: Aerobicrespiration

NADH dehydrogenase I chain E

lethal

b2286

nuoC

NADH:ubiquinone oxidoreductase, fused CDsubunit

enzyme; Energy metabolism, carbon: Aerobicrespiration

NADH dehydrogenase I chain C, D

lethal

b2287

nuoB

NADH:ubiquinone oxidoreductase, chain B

enzyme; Energy metabolism, carbon: Aerobicrespiration

NADH dehydrogenase I chain B

lethal

b2288

nuoA

NADH:ubiquinone oxidoreductase, membrane subunitA

enzyme; Energy metabolism, carbon: Aerobicrespiration

NADH dehydrogenase I chain A

lethal

b2415

ptsH

phosphohistidinoprotein-hexosephosphotransferase component of PTS system (Hpr)

enzyme; Transport of small molecules:Carbohydrates, organic acids, alcohols

PTS system protein HPr

lethal

b2416

ptsI

PEP-protein phosphotransferase of PTS system(enzyme I)

enzyme; Transport of small molecules:Carbohydrates, organic acids, alcohols

PEP-protein phosphotransferase system enzyme I

lethal

b2779

eno

enolase

enzyme; Energy metabolism, carbon: Glycolysis

 

lethal

b2926

pgk

phosphoglycerate kinase

enzyme; Energy metabolism, carbon: Glycolysis

 

lethal

b3236

mdh

malate dehydrogenase, NAD(P)-binding

enzyme; Energy metabolism, carbon: TCA cycle

malate dehydrogenase

lethal

b3731

atpC

F1 sector of membrane-bound ATP synthase,epsilon subunit

enzyme; ATP-proton motive forceinterconversion

membrane-bound ATP synthase, F1 sector,epsilon-subunit

lethal

b3732

atpD

F1 sector of membrane-bound ATP synthase, betasubunit

enzyme; ATP-proton motive forceinterconversion

membrane-bound ATP synthase, F1 sector,beta-subunit

lethal

b3733

atpG

F1 sector of membrane-bound ATP synthase, gammasubunit

enzyme; ATP-proton motive forceinterconversion

membrane-bound ATP synthase, F1 sector,gamma-subunit

lethal

b3734

atpA

F1 sector of membrane-bound ATP synthase, alphasubunit

enzyme; ATP-proton motive forceinterconversion

membrane-bound ATP synthase, F1 sector,alpha-subunit

lethal

b3735

atpH

F1 sector of membrane-bound ATP synthase, deltasubunit

enzyme; ATP-proton motive forceinterconversion

membrane-bound ATP synthase, F1 sector,delta-subunit

lethal

b3736

atpF

F0 sector of membrane-bound ATP synthase,subunit b

enzyme; ATP-proton motive forceinterconversion

membrane-bound ATP synthase, F0 sector, subunit b

lethal

b3737

atpE

F0 sector of membrane-bound ATP synthase,subunit c

enzyme; ATP-proton motive forceinterconversion

membrane-bound ATP synthase, F0 sector, subunit c

lethal

b3738

atpB

F0 sector of membrane-bound ATP synthase,subunit a

enzyme; ATP-proton motive forceinterconversion

membrane-bound ATP synthase, F0 sector, subunit a

lethal

b3919

tpiA

triosephosphate isomerase

enzyme; Energy metabolism, carbon: Glycolysis

 

lethal

b3956

ppc

phosphoenolpyruvate carboxylase

enzyme; Energy metabolism, carbon:Fermentation

 

lethal

b4025

pgi

glucosephosphate isomerase

enzyme; Energy metabolism, carbon: Glycolysis

 

lethal



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 349
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 765
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
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