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Literature summary for 1.1.1.331 extracted from

  • Kuo, H.J.; Wei, Z.Y.; Lu, P.C.; Huang, P.L.; Lee, K.T.
    Bioconversion of pinoresinol into matairesinol by use of recombinant Escherichia coli (2014), Appl. Environ. Microbiol., 80, 2687-2692 .
    View publication on PubMedView publication on EuropePMC
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Cloned(Commentary)

Cloned (Comment) Organism
gene sdh-PpH, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli strain M15. Genes plr-PpH, encoding pinoresinol-lariciresinol reductase (PLR), and sdh-PpH are linked to form two bifunctional fusion genes, plr-sdh and sdh-plr, which are expressed as functional His-tagged proteins in Escherichia coli strain M15. Establishment of a high-efficiency system for the conversion of (+)-pinoresinol to (-)-matairesinol in Escherichia coli expressing a Sdh-PpH fusion protein Dysosma pleiantha

Protein Variants

Protein Variants Comment Organism
additional information the two genes, termed plr-PpH and sdh-PpH, encoding pinoresinol-lariciresinol reductase (PLR) and secoisolariciresinol dehydrogenase (SDH), are linked to form two bifunctional fusion genes, plr-sdh and sdh-plr, which are expressed in Escherichia coli and purified. The proteins are linked via a (GGGGS)4 protein linker to maintain flexibility. Bioconversion in vitro at 22°C for 60 min shows that the conversion efficiency of fusion protein PLR-SDH is higher than that of the mixture of recombinant PLR and reacombinant SDH. The percent conversion of (+)-pinoresinol to matairesinol is 49.8% using PLR-SDH and only 17.7% using a mixture of rPLR and rSDH. Conversion of (+)-pinoresinol by fusion protein SDH-PLR stops at the intermediate product, secoisolariciresinol. In vivo, (+)-pinoresinol is completely converted to matairesinol by living recombinant Escherichia coli expressing PLR-SDH without addition of cofactors Dysosma pleiantha

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.231
-
 (-)-secoisolariciresinol pH 8.8, 20°C, recombinant His-tagged enzyme Dysosma pleiantha

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(-)-secoisolariciresinol + 2 NAD+ Dysosma pleiantha
-
 (-)-matairesinol + 2 NADH + 2 H+
-
 ?

Organism

Organism UniProt Comment Textmining
Dysosma pleiantha W8NQ72 i.e Dysosma pleiantha
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme SDH and bifunctional fusion enzyme PLR-SDH from Escherichia coli Dysosma pleiantha

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(-)-secoisolariciresinol + 2 NAD+
-
 Dysosma pleiantha (-)-matairesinol + 2 NADH + 2 H+
-
 ?

Subunits

Subunits Comment Organism
? x * 34000, recombinant His-tagged enzyme, SDS-PAGE, x * 65000, recombinant His-tagged fusion enzyme PLR-SDH , SDS-PAGE Dysosma pleiantha

Synonyms

Synonyms Comment Organism
SDH
-
 Dysosma pleiantha
sdh-PpH
-
 Dysosma pleiantha
secoisolariciresinol dehydrogenase
-
 Dysosma pleiantha

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
20
-
 recombinant His-tagged Sdh Dysosma pleiantha
22
-
 recombinant His-tagged fusion enzyme Sdh-PpH Dysosma pleiantha

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 recombinant His-tagged fusion enzyme Sdh-PpH Dysosma pleiantha
8.8
-
 recombinant His-tagged Sdh Dysosma pleiantha

Cofactor

Cofactor Comment Organism Structure
NAD+ the enzyme contains a conserved NAD+ binding domain, GxGGxG Dysosma pleiantha