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Literature summary for 1.97.1.4 extracted from

  • Zhang, Y.; Dai, Z.; Krivoruchko, A.; Chen, Y.; Siewers, V.; Nielsen, J.
    Functional pyruvate formate lyase pathway expressed with two different electron donors in Saccharomyces cerevisiae at aerobic growth (2015), FEMS Yeast Res., 15, fov024 .
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene pflA, coexpression of pyruvate formate lyase (PFL, gene pflB, UniProt ID P09373 ) with the pyruvate formate lyase activating enzyme, as well as with appropriate electron donors flavodoxin and ferredoxin (encoded by genes fldA and fdx, respectively), in a non-ethanol-producing Saccharomyces cerevisiae strain IMI076 lacking pyruvate decarboxylase and having a reduced glucose uptake rate due to a mutation in the transcriptional regulator Mth1, IMI076 (Pdc- MTH1-DELTAT ura3-52), plasmid maps, overview. Reduced flavodoxin is the preferred electron donor for PFL, but coexpression of either of the electron donors has a positive effect on growth under aerobic conditions. Subcloning in Escherichia coli strain DH5alpha. The PFL pathway can be functional at aerobic growth conditions in yeast when coexpressed with appropriate electron donors Escherichia coli

Engineering

Protein Variants Comment Organism
additional information recombinant coexpression of the enzyme with pyruvate format lyase and flavodoxin or ferredoxin in Saccharomyces cerevisiae leads to over 20fold increased expression of endogenous formate dehydrogenases FDH1 and FDH2 Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
S-adenosyl-L-methionine + dihydroflavodoxin + [formate C-acetyltransferase]-glycine Escherichia coli
-
5'-deoxyadenosine + L-methionine + flavodoxin semiquinone + [formate C-acetyltransferase]-glycin-2-yl radical
-
?

Organism

Organism UniProt Comment Textmining
Escherichia coli P0A9N4
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information PFL-AE utilizes S-adenosylmethionine (SAM) and reduced flavodoxin as the cosubstrates to generate a 5'-deoxyadenosyl radical, which then activates PFL by abstracting a hydrogen atom from residue G734 Escherichia coli ?
-
?
S-adenosyl-L-methionine + dihydroflavodoxin + [formate C-acetyltransferase]-glycine
-
Escherichia coli 5'-deoxyadenosine + L-methionine + flavodoxin semiquinone + [formate C-acetyltransferase]-glycin-2-yl radical
-
?

Synonyms

Synonyms Comment Organism
PFL-AE
-
Escherichia coli
PflA
-
Escherichia coli
pyruvate formate lyase activating enzyme
-
Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
Ferredoxin can partly substitute for flavodoxin Escherichia coli
flavodoxin preferred cofactor Escherichia coli
additional information reduced flavodoxin serves as an electron donor and SAM as a cosubstrate for PFL-AE to generate a 5'-deoxyadenosyl radical, which is responsible for PFL activation Escherichia coli
S-adenosyl-L-methionine
-
Escherichia coli