Jonathan WorrallMCB away day

25.09.13

An appraisal of metals used in proteins

Systematic survey of 1371 different enzymes with a known 3D structure

47% required metals with 41% requiring metals at their catalytic sites

JBIC (2008) 13, 1205-1218Nature (2009) 460, 823-830

Research Interests

Microbial biotechnologyApoptosis

Cytochrome c/phospholipid complexes Streptomyces lividans

Cu

Worrall & Vijgenboom 2010, Nat. Prod. Reps. 5, 742.

Cuproproteins and enzymes must havea role in triggering development switches

R580 hours

R5 10 μM Cu(II)80 hours

S. lividans

R5140 hours

ΔSLI4214

ΔSLI4213

ΔSLI4212

ΔSLI3188

DCuproproteins/enzymes have morphological phenotypes

Blundell et al. 2013, Open Biology 3, 120163

Ty act / biomass

-500

0

500

1000

1500

2000

2500

3000

3500

4000

0 20 40 60

Time (hour)Ty

act

ivity

(uni

ts/m

in,g

,lit)

1326 pIJ703

2837::Apra pIJ703

DSLI3188

S. lividans

Morphological Engineering

Reengineering of Streptomyces strains as hosts for synthetic biology approaches to create sustainable industrial production platforms

Techniques used

• Molecular biology• Recombinant protein over-expression • Protein chromatography• UV-visible, circular dichroism (CD) spectroscopies • Electron paramagnetic resonance (EPR) spectroscopy• Nuclear magnetic resonance (NMR) spectroscopy (UEA)• X-ray crystallography• Isothermal titration calorimetry (ITC)• Stopped-flow kinetics• Electrophoretic mobility shift assays (EMSA)

Collaborations

UEA (Cytochrome c)Professor Geoff Moore (NMR)

Professor Julea Butt (Electrochemistry)

Molecular Biotechnology, Leiden, NLDr Erik Vijgenboom

EssexDr Mike Hough (X-ray)

Dr Dimitri Svistunenko (EPR)Professor Mike Wilson (SF-kinetics)