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Literature summary extracted from

  • Kittendorf, J.D.; Beck, B.J.; Buchholz, T.J.; Seufert, W.; Sherman, D.H.
    Interrogating the molecular basis for multiple macrolactone ring formation by the pikromycin polyketide synthase (2007), Chem. Biol., 14, 944-954.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.1.239 overexpression of pikromycin modules PikAIII and PIkAIV in Escherichia coli Streptomyces venezuelae
2.3.1.240 overexpression of pikromycin modules PikAIII and PIkAIV in Escherichia coli Streptomyces venezuelae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.1.239 additional information
-
 additional information both PikAIV(DELTAAT) and PikAIV(DELTAACP) display similar apparent kcat values for 10-deoxymethynolide production, which are approximately 2fold lower than the calculated kcat from wild-type PikAIII and PikAIV. Likewise, both PikAIV mutants display similar apparent KM values that are also reduced 2fold when compared to the KM from PikAIII and PikAIV. The equivalent decrease in the apparent kinetic parameters for both mutants results in a specificity constant (kcat/KM) that is comparable to wild-type, further emphasizing the non-critical role of these catalytic domains in 10-deoxymethynolide production. Both PikAIII(DELTADock) (partial deletions of PikAIII C-terminal domain) and PikAIV(DELTADock) (partial deletion of PikAIV N-terminal docking domains) display apparent kcat values (0.044/min and 0.053/min), which are significantly decrease relative to the kcat observed with wild-type PikAIII and PikAIV. The apparent KM values for PikAIII(DELTADock) and PikAIV (DELTADock) decrease 3–5 fold compared to wild-type, resulting in specificity constants (kcat/KM) that are 14fold and 8fold lower, respectively. Combining PikAIII(DELTADock) and PikAIV(DELTADock) together do not result in an additional rate decrease. The kinetic parameters are comparable to those obtained when either truncated monomodule is paired with its wild-type partner Streptomyces venezuelae
2.3.1.240 additional information
-
 additional information both PikAIV(DELTAAT) and PikAIV(DELTAACP) display similar apparent kcat values for 10-deoxymethynolide production, which are approximately 2fold lower than the calculated kcat from wild-type PikAIII and PikAIV. Likewise, both PikAIV mutants display similar apparent KM values that are also reduced 2fold when compared to the KM from PikAIII and PikAIV. The equivalent decrease in the apparent kinetic parameters for both mutants results in a specificity constant (kcat/KM) that is comparable to wild-type, further emphasizing the non-critical role of these catalytic domains in 10-deoxymethynolide production. Both PikAIII(DELTADock) (partial deletions of PikAIII C-terminal domain) and PikAIV(DELTADock) (partial deletion of PikAIV N-terminal docking domains) display apparent kcat values (0.044/min and 0.053/min), which are significantly decrease relative to the kcat observed with wild-type PikAIII and PikAIV. The apparent KM values for PikAIII(DELTADock) and PikAIV (DELTADock) decrease 3–5 fold compared to wild-type, resulting in specificity constants (kcat/KM) that are 14fold and 8fold lower, respectively. Combining PikAIII(DELTADock) and PikAIV(DELTADock) together do not result in an additional rate decrease. The kinetic parameters are comparable to those obtained when either truncated monomodule is paired with its wild-type partner Streptomyces venezuelae

Organism

EC Number Organism UniProt Comment Textmining
2.3.1.239 Streptomyces venezuelae Q9ZGI5 and Q9ZGI4 and Q9ZGI3 and Q9ZGI2 and Q9ZGI1 Q9ZGI5: type I polyketide synthase PikAI, Q9ZGI4: type I polyketide synthase PikAII, Q9ZGI3: type I polyketide synthase PikAIII, Q9ZGI2: type I polyketide synthase PikAIV, Q9ZGI1: thioesterase II PikAV. Multifunctional, modular enzyme
-
2.3.1.240 Streptomyces venezuelae Q9ZGI5 and Q9ZGI4 and Q9ZGI3 and Q9ZGI2 and Q9ZGI1 Q9ZGI5: type I polyketide synthase PikAI, Q9ZGI4: type I polyketide synthase PikAII, Q9ZGI3: type I polyketide synthase PikAIII, Q9ZGI2: type I polyketide synthase PikAIV, Q9ZGI1: thioesterase II PikAV. Multifunctional, modular enzyme
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.1.239
-
 Streptomyces venezuelae
2.3.1.240
-
 Streptomyces venezuelae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.1.239 (E)-(2S,4R,8R,9R)-S-2-acetamidoethyl 9-hydroxy-2,4,8-trimethyl-5-oxoundec-6-enethioate + (2S)-methylmalonyl-CoA + NADPH + H+ pikromycin modules PikAIII and PikAIV generate the 12-membered ring macrocycle most efficiently when engaged in their native protein-protein interaction. PikAIV adopts an alternative conformation that enables the terminal thioesterase domain to directly off-load the PikAIII-bound hexaketide intermediate for macrocyclization Streptomyces venezuelae 10-deoxymethynolide + ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.3.1.239 pikromycin polyketide synthase
-
 Streptomyces venezuelae
2.3.1.240 pikromycin polyketide synthase
-
 Streptomyces venezuelae

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.3.1.239 30
-
 assay at Streptomyces venezuelae
2.3.1.240 30
-
 assay at Streptomyces venezuelae

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.1.239 7.2
-
 assay at Streptomyces venezuelae
2.3.1.240 7.2
-
 assay at Streptomyces venezuelae