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signtruculentBiotechnology

Oct 2, 2013 (3 years and 8 months ago)

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Predicting

and testing
enzymatic activity

Charlie Harrison

Supervisors: Profs. John Ward &
Christine
Orengo


Norcoclaurine


Benzylisoquinoline

alkaloids (
BIQAs
)


group of plant secondary metabolites




I
ncludes

many interesting compounds


Common entry point:
norcoclaurine

(S)
-
norcoclaurine

(R)
-
norcoclaurine

Norcoclaurine


N
orco
C
laurine

S
ynthase



NCS


Five known examples from three different
species


But
BIQAs

exist in many more species…


Can we find more
NCSs
?

Main Project Aims


Bioinformatics


Identify
uncharacterised

proteins that are likely to
exhibit NCS activity


Lab Work


Express these proteins in
E.coli

and test
predictions

Bioinformatics


Available data:
sequence
and
structure


Use these to predict function

Bioinformatics

Reproduced from
Orengo
, Jones &
Thornton
2003
with permission

Bioinformatics


Structural data is better for identifying
functional homologues...

Bioinformatics


Structural data is better for identifying
functional homologues...but is only available
for ~1% of sequences


Use predictions of structural regions


Gene3D


Starting point: Gene3D family of likely
structural relatives


Sequence clustering

Bioinformatics

Lab Work


Design and order the synthetic genes





Wait 3 weeks…

Lab Work

Results







Results

Thanks