EG10804

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Author Information

Anjini Karthik

Basic Information

  • ID: EG10804
  • Name: pykF
  • Organism: E. coli
  • UniProt ID: P0AD61 [1]
  • Description: The gene pykF codes for pyruvate kinase I, an enzyme that is essential for glycolysis, a pathway that generates ATP. The formation of pyruvate and ATP with the help of pyruvate kinase I is the last and crucial step in the pathway and is irreversible under physiological conditions. This gene works with a sister gene, pykA, which codes for pyruvate kinase II; but the two enzymes are not interchangeable, as they are activated by different substrates. Pyruvate kinase I has demonstrated activation by fructose and is often targeted in metabolic engineering due to the key role it plays in controlling metabolic flux in glycolysis [[2]]. Thus, this gene must function for cellular respiration to occur, the main method for a cell to extract and use energy.
  • DNA Length: 1413 base pairs.
  • DNA sequence:

ATG AAA AAA ACA AAA ATT GTC TGC ACG ATT GGT CCC AAG ACT GAA TCT GAA GAG ATG TTG GCT AAG ATG CTT GAT GCG GGT ATG AAC GTC ATG CGT TTA AAT TTC TCT CAT GGT GAT TAC GCC GAG CAC GGT CAG CGC ATC CAG AAT CTG CGC AAC GTG ATG AGT AAG ACT GGC AAG ACC GCT GCT ATT CTT TTA GAC ACG AAA GGG CCA GAA ATC CGC ACC ATG AAA CTG GAA GGG GGA AAC GAT GTC AGC TTA AAG GCA GGT CAG ACC TTT ACC TTC ACC ACT GAC AAG AGT GTG ATT GGG AAT TCG GAA ATG GTT GCA GTG ACA TAT GAG GGT TTT ACG ACA GAC TTA TCC GTC GGT AAC ACC GTG TTG GTC GAT GAT GGC TTA ATC GGA ATG GAA GTC ACT GCA ATC GAA GGT AAT AAA GTC ATC TGT AAG GTA CTT AAC AAC GGG GAT TTG GGC GAG AAT AAG GGT GTC AAT TTG CCG GGA GTA AGC ATC GCG CTG CCT GCA CTG GCT GAA AAA GAC AAA CAA GAT TTA ATT TTC GGC TGC GAG CAG GGT GTG GAT TTC GTA GCA GCC TCG TTC ATC CGT AAA CGT TCA GAT GTC ATC GAA ATC CGC GAA CAT CTG AAA GCC CAT GGA GGC GAG AAC ATT CAT ATC ATT TCT AAG ATC GAA AAT CAG GAG GGA TTA AAT AAC TTC GAC GAG ATT TTA GAG GCA AGT GAT GGT ATT ATG GTG GCG CGC GGC GAT TTG GGT GTC GAG ATT CCT GTA GAG GAG GTT ATC TTT GCA CAG AAG ATG ATG ATC GAA AAG TGT ATT CGC GCA CGT AAG GTG GTT ATC ACT GCG ACC CAA ATG CTT GAT TCC ATG ATT AAG AAC CCA CGT CCA ACA CGC GCT GAG GCA GGC GAC GTT GCG AAT GCC ATC TTA GAC GGC ACA GAT GCC GTG ATG CTG AGT GGA GAG AGT GCA AAG GGG AAA TAC CCT TTA GAA GCA GTC AGT ATC ATG GCA ACT ATC TGT GAA CGC ACC GAT CGT GTA ATG AAC TCA CGT TTG GAA TTT AAC AAC GAC AAC CGT AAA CTG CGC ATT ACG GAA GCG GTA TGC CGT GGC GCG GTC GAA ACT GCC GAA AAA CTT GAT GCG CCC TTG ATT GTT GTT GCA ACA CAG GGT GGC AAG TCC GCA CGC GCA GTC CGT AAG TAT TTC CCC GAC GCA ACG ATT CTG GCG CTT ACC ACA AAC GAA AAG ACA GCA CAT CAA CTG GTA TTA TCG AAG GGC GTT GTT CCG CAA TTA GTA AAG GAG ATT ACA TCA ACC GAT GAC TTC TAT CGC TTG GGC AAG GAA CTT GCA CTG CAA AGT GGT CTT GCA CAC AAG GGG GAT GTC GTG GTA ATG GTG AGT GGC GCC CTG GTC CCT TCA GGA ACG ACG AAC ACA GCA TCT GTG CAC GTC TTA TAA

  • Amino Acid length: 470 amino acids.
  • Amino Acid sequence:

MKKTKIVCTI GPKTESEEML AKMLDAGMNV MRLNFSHGDY AEHGQRIQNL RNVMSKTGKT AAILLDTKGP EIRTMKLEGG NDVSLKAGQT FTFTTDKSVI GNSEMVAVTY EGFTTDLSVG NTVLVDDGLI GMEVTAIEGN KVICKVLNNG DLGENKGVNL PGVSIALPAL AEKDKQDLIF GCEQGVDFVA ASFIRKRSDV IEIREHLKAH GGENIHIISK IENQEGLNNF DEILEASDGI MVARGDLGVE IPVEEVIFAQ KMMIEKCIRA RKVVITATQM LDSMIKNPRP TRAEAGDVAN AILDGTDAVM LSGESAKGKY PLEAVSIMAT ICERTDRVMN SRLEFNNDNR KLRITEAVCR GAVETAEKLD APLIVVATQG GKSARAVRKY FPDATILALT TNEKTAHQLV LSKGVVPQLV KEITSTDDFY RLGKELALQS GLAHKGDVVV MVSGALVPSG TTNTASVHVL*

Function and Homologs

  • Product: Pyruvate kinase I
  • Closest homologous proteins: The top (max three) homologous proteins to this protein, as identified by BLAST searches.
    • Pyruvate kinase I (Proteobacteria), 956/100%/0.0/100%, [3]
    • Pyruvate kinase (Enterobacteriacaea), 954/100%/0.0/100%, [4]
    • Pyruvate kinase II (Proteobacteria), 280/99%/1e-87/37%, [5]
  • Equivalent E. coli / JCVI functional protein: MMSYN1_0221.

Expression

  • Expression Level: High
  • Expression Level Hypothesis: The expression level of pykF is high in E. coli, as opposed to low in its homolog. This phenomenon could be observed as a result of the fact that E. coli depends on many different sources of fuel (it can grow using many different compounds), and pykF is more versatile with fuel sources that aren't glucose.
  • Expression Level References and Description: This information is based on the E. coli Proteomic Expression dataset.
  • Expression Time: Early to middle
  • Expression Level Hypothesis: pykF is not necessary until the organism has already taken in a source of fuel (such as a sugar). Intake must happen first. However, glycolysis is the first step in the photosynthetic pathway, so this gene overall should be expressed in the early to middle stage.
  • Expression Time References and Description: Resources used were the same as those cited in the “Basic Information/Description” section.

Gene Context

  • Other Components: pykA, which codes for pyruvate kinase II, is one other gene needed for the module to work— there was no explicit E. coli pykA gene found in this database.
  • Possible Dependencies: hprK: Sugars must be present along with molecules like ADP to enable pyruvate kinase to convert the sugar source into pyruvate and energy in the form of ATP.
  • Process: Cellular respiration
    • Inputs: Fructose 1,6-bisphosphate, ADP
    • Outputs: Pyruvate, ATP
    • Resources: Resources used were the same as those cited in the “Basic Information/Description” section.

Construct

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  • Synthesis Score: The synthesis score: 1, 2,3
  • Predicted Translation Rate: Prediction of construct translation rate from the RBS calculator
  • Design Notes and Details:
  • GenBank File: A link to the GenBank file. file