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Literature summary extracted from

  • Schmidt, T.; Lenders, M.; Hillebrand, A.; van Deenen, N.; Munt, O.; Reichelt, R.; Eisenreich, W.; Fischer, R.; Pruefer, D.; Gronover, C.S.
    Characterization of rubber particles and rubber chain elongation in Taraxacum kok-saghyz (2010), BMC Biochem., 11, 11.
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
2.5.1.20 molecular biology model for rubber biosynthesis Taraxacum kok-saghyz

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.5.1.20 expression in Saccharomyces cerevisiae SNH23-D7 (temperature sensitive mutant strain for dehydrodoichyl diphosphate synthase) or Nicotiana tabacum protoplasts, enzyme remains fully functional, and expression in Escherichai coli BL21 for sequencing and Western blot analysis with antibodies Taraxacum kok-saghyz

Protein Variants

EC Number Protein Variants Comment Organism
2.5.1.20 additional information complementation of enzyme deficient Saccharomyces by each Taraxacum kok-saghyz rubber transferase enzyme, protein extracts from protoplasts expressing recombinant enzyme show high activities of each isozyme Taraxacum kok-saghyz

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.5.1.20 Mg2+ 2 to 5 mM MgCl2 increase 2-methyl-1,3-butadiene incorporation, optimum 4-5 mM, with 7.5 mM decline below the 2 mM value, almost loss of activity with 10 mM MgCl2 Taraxacum kok-saghyz
2.5.1.20 Mg2+ increase in in 2-methyl-1,3-butadiene incorporation from 2 up to 5 mM MgCl2, optimum 4-5 mM, with 7.5 mM decline below the 2 mM value, almost loss of activity with 10 mM MgCl2 Taraxacum kok-saghyz

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.5.1.20 32000
-
 SDS-PAGE, TkCPT1 Taraxacum kok-saghyz
2.5.1.20 32000
-
 SDS-PAGE, TkCPT2 Taraxacum kok-saghyz
2.5.1.20 34000
-
 SDS-PAGE, TkCPT3 Taraxacum kok-saghyz

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.5.1.20 additional information Taraxacum kok-saghyz natural rubber is synthesized by adding activated 2-methyl-1,3-butadiene to the growing chain ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.20 Taraxacum kok-saghyz
-
 Russian dandelion
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.5.1.20 purification of rubber particles containing the enzyme by harvesting latex from petioles of roots, separating latex fractions by centrifugation with extraction buffer (100 mM Tris-HCl, pH 7.8, 350 mM sorbitol, 10 mM NaCl, 5 mM MgCl2, 5 mM DTT), rubber phase is further centrifuged, washed with buffer and used for enzyme assays Taraxacum kok-saghyz

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.5.1.20 latex
-
 Taraxacum kok-saghyz
-
2.5.1.20 additional information only minimally expressed in leaves, pedicels, and roots Taraxacum kok-saghyz
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.5.1.20 0.0000026
-
 2-methyl-1,3-butadiene incorporation assay with rubber particles, 100 mM Tris-HCl, pH 7.5, 30°C, 2 h Taraxacum kok-saghyz
2.5.1.20 0.000003
-
 2-methyl-1,3-butadiene incorporation assay with rubber particles, 100 mM Tris-HCl, pH 7.5, 30°C, 2 h Taraxacum kok-saghyz

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.20 isopentenyl diphosphate + (E,E)-farnesyl diphosphate + cis-1,4-polyisoprene
-
 Taraxacum kok-saghyz diphosphate + a poly-cis-polyprenyl diphosphate longer by one C5 unit
-
 ?
2.5.1.20 additional information natural rubber is synthesized by adding activated 2-methyl-1,3-butadiene to the growing chain Taraxacum kok-saghyz ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.5.1.20 cis-1,4-polyprenylcistransferase
-
 Taraxacum kok-saghyz
2.5.1.20 cis-prenyltransferase
-
 Taraxacum kok-saghyz
2.5.1.20 rubber transferase
-
 Taraxacum kok-saghyz
2.5.1.20 TkCPT1
-
 Taraxacum kok-saghyz
2.5.1.20 TkCPT2
-
 Taraxacum kok-saghyz
2.5.1.20 TkCPT3
-
 Taraxacum kok-saghyz

General Information

EC Number General Information Comment Organism
2.5.1.20 metabolism enzyme produces very pure poly(cis-1,4-isoprene) and retains its capacity to produce natural rubber ex vivo Taraxacum kok-saghyz