Broadening substrate specificity of a chain-extending ketosynthase through a single active-site mutation.
An in vitro model system based on a ketosynthase domain of the erythromycin polyketide synthase was used to probe the apparent substrate tolerance of ketosynthase domains of the mycolactone polyketide synthase. A specific residue change was identified that led to an emphatic increase in turnover of a range of substrates.
Department of Biochemistry, University of Cambridge, Sanger Building, 80 Tennis Court Road, Cambridge CB2 1GA, UK. acm95@cam.ac.uk pfl10@cam.ac.uk.
Annabel C
AC
Murphy
Department of Biochemistry, University of Cambridge, Sanger Building, 80 Tennis Court Road, Cambridge CB2 1GA, UK. acm95@cam.ac.uk pfl10@cam.ac.uk.
Hui
H
Hong
Department of Biochemistry, University of Cambridge, Sanger Building, 80 Tennis Court Road, Cambridge CB2 1GA, UK. acm95@cam.ac.uk pfl10@cam.ac.uk and Crescendo Biologics Ltd, Meditrina Building 260, Babraham Research Campus, Cambridge CB22 3AT, UK.
Steve
S
Vance
Department of Biochemistry, University of Cambridge, Sanger Building, 80 Tennis Court Road, Cambridge CB2 1GA, UK. acm95@cam.ac.uk pfl10@cam.ac.uk.
R William
RW
Broadhurst
Department of Biochemistry, University of Cambridge, Sanger Building, 80 Tennis Court Road, Cambridge CB2 1GA, UK. acm95@cam.ac.uk pfl10@cam.ac.uk.
Peter F
PF
Leadlay
D020134:Catalytic Domain; D009154:Mutation; D048630:Polyketide Synthases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering; D013379:Substrate Specificity
D048630:Polyketide Synthases