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Literature summary for 4.2.3.150 extracted from

  • Tomita, T.; Kim, S.Y.; Teramoto, K.; Meguro, A.; Ozaki, T.; Yoshida, A.; Motoyoshi, Y.; Mori, N.; Ishigami, K.; Watanabe, H.; Nishiyama, M.; Kuzuyama, T.
    Structural insights into the CotB2-catalyzed cyclization of geranylgeranyl diphosphate to the diterpene cyclooctat-9-en-7-ol (2017), ACS Chem. Biol., 12, 1621-1628 .
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene cotB2, recombinant expression of His-tagged mutant enzymes in Escherichia coli strain BL21(DE3) Streptomyces melanosporofaciens

Protein Variants

Protein Variants Comment Organism
F107A site-directed mutagenesis, the mutant follows a different reaction mechanism compared to wild-type and produces the different product cembrane A instead of cyclooctat-9-en-7-ol Streptomyces melanosporofaciens
additional information mutational analysis of the atypical aspartate-rich motif of CotB2 (EC 4.2.3.146). The substrate specificity of the CotB2 mutants can be completely changed. Proposed cyclization mechanism for CotB2 and its mutants, overview Streptomyces melanosporofaciens
W186L site-directed mutagenesis, the mutant follows a different reaction mechanism compared to wild-type and produces the different products cembrane A and 3,7,18-dolabellatriene and only low amounts of cyclooctat-9-en-7-ol Streptomyces melanosporofaciens

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Streptomyces melanosporofaciens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
geranylgeranyl diphosphate Streptomyces melanosporofaciens
-
cembrene A + diphosphate
-
?
geranylgeranyl diphosphate Streptomyces melanosporofaciens MI614-43F2
-
cembrene A + diphosphate
-
?

Organism

Organism UniProt Comment Textmining
Streptomyces melanosporofaciens
-
-
-
Streptomyces melanosporofaciens MI614-43F2
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Streptomyces melanosporofaciens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
geranylgeranyl diphosphate
-
Streptomyces melanosporofaciens cembrene A + diphosphate
-
?
geranylgeranyl diphosphate activity of enzyme mutants F107A and W186L, not of wild-type enzyme Streptomyces melanosporofaciens cembrene A + diphosphate
-
?
geranylgeranyl diphosphate
-
Streptomyces melanosporofaciens MI614-43F2 cembrene A + diphosphate
-
?
geranylgeranyl diphosphate activity of enzyme mutants F107A and W186L, not of wild-type enzyme Streptomyces melanosporofaciens MI614-43F2 cembrene A + diphosphate
-
?
additional information the mutants F107A and W186L follow a different reaction mechanism compared to wild-type CotB2, and produce the different product cembrane A Streptomyces melanosporofaciens ?
-
?
additional information the mutants F107A and W186L follow a different reaction mechanism compared to wild-type CotB2, and produce the different product cembrane A Streptomyces melanosporofaciens MI614-43F2 ?
-
?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Streptomyces melanosporofaciens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Streptomyces melanosporofaciens