Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 4.2.3.30 extracted from

  • Jia, M.; Zhou, K.; Tufts, S.; Schulte, S.; Peters, R.J.
    A pair of residues that interactively affect diterpene synthase product outcome (2017), ACS Chem. Biol., 12, 862-867 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 4.2.3.30

Cloned(Commentary)

Cloned (Comment) Organism
sequence comparisons and phylogenetic analysis and tree Oryza sativa Japonica Group

Protein Variants

Protein Variants Comment Organism
additional information of the three differences in active site residues between the orthologous OsKSL5j and OsKSL5i, switching the Ile at position 664 in OsKSL5i to the Thr found in OsKSL5j is sufficient to short circuit the production of ent-isokaurene, as OsKSL5i:I664T specifically produces ent-pimara-8(14),15-diene instead. But the reciprocal residue switch leads to production of a mixture of tetracycles ent-isokaurene, ent-kaurene, and ent-atiserene (i.e. by OsKSL5j:T664I), rather than the almost exclusive production of ent-isokaurene observed with OsKSL5i Oryza sativa Japonica Group
T664I site-directed mutagenesis, the OsKSL5j:T664I mutant produces only ent-pimara-8(14),15-diene Oryza sativa Japonica Group
T664I/L661V site-directed mutagenesis, the addition of L661Vto mutation T664I leads to much more specific production of ent-isokaurene, the product outcome mediated by this OsKSL5j:L661V/T664I double residue switch mutant is essentially identical to that mediated by OsKSL5i Oryza sativa Japonica Group
T664I/V718I site-directed mutagenesis, the addition of V718I to mutation T664I does not further alter product outcome Oryza sativa Japonica Group

Localization

Localization Comment Organism GeneOntology No. Textmining
chloroplast the enzyme contains an N-terminal transit peptide sequence Oryza sativa Japonica Group 9507
-

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Oryza sativa Japonica Group

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ent-copalyl diphosphate Oryza sativa Japonica Group
-
 ent-pimara-8(14),15-diene + diphosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Oryza sativa Japonica Group Q6Z5J6
-
-

Source Tissue

Source Tissue Comment Organism Textmining
leaf
-
 Oryza sativa Japonica Group
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ent-copalyl diphosphate
-
 Oryza sativa Japonica Group ent-pimara-8(14),15-diene + diphosphate
-
 ?
additional information GC-MS reaction product analysis. The distinct function of two alleles of a KSL from rice, OsKSL5i and OsKSLj, identifies a single residue switch with a profound effect on not only OsKSL5 product outcome but also that of land plant KSs more broadly, specifically, replacement of a key isoleucine with threonine, which interrupts formation of the tetracyclic ent-isokaurene at the tricyclic stage, leading to production of ent-pimaradiene instead Oryza sativa Japonica Group ?
-
 ?

Synonyms

Synonyms Comment Organism
kaurene synthase-like enzyme 5
-
 Oryza sativa Japonica Group
KS-like 5j
-
 Oryza sativa Japonica Group
OsKSL5j
-
 Oryza sativa Japonica Group

General Information

General Information Comment Organism
evolution in the plant kingdom, where the labdane-related diterpenoids (LRDs) are predominantly found, the class I and II diterpene cyclases seem to have most directly evolved via gene duplication and neofunctionalization of the ent-kaurene synthases (KSs) required in all vascular plants for gibberellin hormone biosynthesis. Accordingly, these enzymes are often termed KS-like (KSL) and form a distinct subfamily within the plant terpene synthase family. Like class I terpene synthases more generally, the KS(L)s carry out catalysis in a highly conserved alpha-helical bundle domain that contains two signature motifs. These DDxxD and NDxx(S/T)xxxE sequences are involved in ligation of a trinuclear Mg2+ cluster to promote substrate binding and the subsequent initiating ionization of the allylic diphosphate ester bond. Ile664 is conserved across all the known plant KSs, and substitution of Thr is found to similarly alter product outcome with the KS from not only rice (OsKS) but also the eudicot Arabidopsis thaliana (AtKS), leading to predominant production of ent-pimara-8(14),15-diene rather than ent-kaur-16-ene. OsKSL5 belongs to an early diverging KSL lineage within monocots and is relatively distantly related to any KSs Oryza sativa Japonica Group
malfunction substitution of Ile for the Thr found at this position in OsKSL5j does not lead to complete specificity for production of ent-isokaur-15-ene, as the resulting OsKSL5j:T664I also produces small amounts of ent-kaur-16-ene, along with more substantial amounts of ent-atiserene, resulting from alternative rearrangement of the ent-beyeran-12-yl+ intermediate formed by the second cyclization step Oryza sativa Japonica Group
additional information conserved residues at positions 661 (secondary) and 664 (primary) are responsible for substrate specificity. Two functionally distinct alleles of Oryza sativa OsKSL5, which react with ent-copalyl diphosphate (ent-copalyl diphosphate), with that from subspecies japonica (OsKSL5j) producing ent-pimara-8(14),15-diene and that from subspecies indica (OsKSL5i) producing ent-(iso)kaur-15-ene instead Oryza sativa Japonica Group