Advanced

Sequence variation determining stereochemistry of a delta-11 desaturase active in moth sex pheromone biosynthesis

Ding, Baojian LU ; Carraher, Colm LU and Löfstedt, Christer LU (2016) In Insect Biochemistry and Molecular Biology 74. p.68-75
Abstract
A Δ11 desaturase from the oblique banded leaf roller moth Choristoneura rosaceana takes the saturated myristic acid and produces a mixture of (E)-11-tetradecenoate and (Z)-11-tetradecenoate with an excess of the Z isomer (35:65). A desaturase from the spotted fireworm moth Choristoneura parallela also operates on myristic acid substrate but produces almost pure (E)-11-tetradecenoate. The two desaturases share 92% amino acid identity and 97% amino acid similarity. There are 24 amino acids differing between these two desaturases. We constructed mutations at all of these positions to pinpoint the sites that determine the product stereochemistry. We demonstrated with a yeast functional assay that one amino acid at the cytosolic... (More)
A Δ11 desaturase from the oblique banded leaf roller moth Choristoneura rosaceana takes the saturated myristic acid and produces a mixture of (E)-11-tetradecenoate and (Z)-11-tetradecenoate with an excess of the Z isomer (35:65). A desaturase from the spotted fireworm moth Choristoneura parallela also operates on myristic acid substrate but produces almost pure (E)-11-tetradecenoate. The two desaturases share 92% amino acid identity and 97% amino acid similarity. There are 24 amino acids differing between these two desaturases. We constructed mutations at all of these positions to pinpoint the sites that determine the product stereochemistry. We demonstrated with a yeast functional assay that one amino acid at the cytosolic carboxyl terminus of the protein (258E) is critical for the Z activity of the C. rosaceana desaturase. Mutating the glutamic acid (E) into aspartic acid (D) transforms the C. rosaceana enzyme into a desaturase with C. parallela-like activity, whereas the reciprocal mutation of the C. parallela desaturase transformed it into an enzyme producing an intermediate 64:36 E/Z product ratio. We discuss the causal link between this amino acid change and the stereochemical properties of the desaturase and the role of desaturase mutations in pheromone evolution. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Desaturase, Stereochemistry, Di-iron center, Histidine rich motif, Mutagenesis, Moth pheromone evolution
in
Insect Biochemistry and Molecular Biology
volume
74
pages
8 pages
publisher
Elsevier
external identifiers
  • Scopus:84968903304
  • WOS:000378450800008
ISSN
1879-0240
DOI
10.1016/j.ibmb.2016.05.002
language
English
LU publication?
yes
id
bbe9e3da-3f13-471a-a893-d0869cc4660e
date added to LUP
2016-05-30 16:39:41
date last changed
2017-01-16 15:01:10
@article{bbe9e3da-3f13-471a-a893-d0869cc4660e,
  abstract     = {A Δ11 desaturase from the oblique banded leaf roller moth <i>Choristoneura rosaceana</i> takes the saturated myristic acid and produces a mixture of (E)-11-tetradecenoate and (Z)-11-tetradecenoate with an excess of the Z isomer (35:65). A desaturase from the spotted fireworm moth <i>Choristoneura parallela</i> also operates on myristic acid substrate but produces almost pure (E)-11-tetradecenoate. The two desaturases share 92% amino acid identity and 97% amino acid similarity. There are 24 amino acids differing between these two desaturases. We constructed mutations at all of these positions to pinpoint the sites that determine the product stereochemistry. We demonstrated with a yeast functional assay that one amino acid at the cytosolic carboxyl terminus of the protein (258E) is critical for the Z activity of the <i>C. rosaceana</i> desaturase. Mutating the glutamic acid (E) into aspartic acid (D) transforms the <i>C. rosaceana</i> enzyme into a desaturase with C. parallela-like activity, whereas the reciprocal mutation of the <i>C. parallela</i> desaturase transformed it into an enzyme producing an intermediate 64:36 E/Z product ratio. We discuss the causal link between this amino acid change and the stereochemical properties of the desaturase and the role of desaturase mutations in pheromone evolution.},
  author       = {Ding, Baojian and Carraher, Colm and Löfstedt, Christer},
  issn         = {1879-0240},
  keyword      = {Desaturase,Stereochemistry,Di-iron center,Histidine rich motif,Mutagenesis,Moth pheromone evolution},
  language     = {eng},
  pages        = {68--75},
  publisher    = {Elsevier},
  series       = {Insect Biochemistry and Molecular Biology},
  title        = {Sequence variation determining stereochemistry of a delta-11 desaturase active in moth sex pheromone biosynthesis},
  url          = {http://dx.doi.org/10.1016/j.ibmb.2016.05.002},
  volume       = {74},
  year         = {2016},
}