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4.2.3.103: ent-isokaurene synthase

This is an abbreviated version!
For detailed information about ent-isokaurene synthase, go to the full flat file.

Word Map on EC 4.2.3.103

Reaction

ent-copalyl diphosphate
=
ent-isokaurene
 +
diphosphate

Synonyms

ent-kaurene synthase like, ent-kaurene synthase like 5, ent-kaurene synthase like 6, kaurene synthase-like, kaurene synthase-like enzyme 5, kaurene synthase-like enzyme 6, KS-like 5i, KS-like 6, KSL5, KSL5i, KSL5ind, KSL6, KSL6ind, OrKSL5ind, OrKSL6, OrKSL6ind, OsKSL5i, OsKSL6

ECTree

     4 Lyases
         4.2 Carbon-oxygen lyases
             4.2.3 Acting on phosphates
                4.2.3.103 ent-isokaurene synthase

Engineering

Engineering on EC 4.2.3.103 - ent-isokaurene synthase

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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
I664T
I718V
the mutant specifically produces ent-isokaur-15-ene from ent-copalyl diphosphate
V661L
I664T
the mutant specifically produces pimara-8(14),15-diene from ent-copalyl diphosphate
V661L
site-directed mutagenesis, substitution of Leu for the Val found at this secondary position only slightly decreases catalytic fidelity, the mutant still predominantly produces ent-isokaurene, with only slight increases in the relative amounts of ent-kaur-16-ene and ent-atiserene
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