title: Insights into 6-Methylsalicylic Acid Bio-assembly by Using Chemical Probes.
abstract: Chemical probes capable of reacting with KS (ketosynthase)-bound biosynthetic intermediates were utilized for the investigation of the model type I iterative polyketide synthase 6-methylsalicylic acid synthase (6-MSAS) in vivo and in vitro. From the fermentation of fungal and bacterial 6-MSAS hosts in the presence of chain termination probes, a full range of biosynthetic intermediates was isolated and characterized for the first time. Meanwhile, in vitro studies of recombinant 6-MSA synthases with both nonhydrolyzable and hydrolyzable substrate mimics have provided additional insights into substrate recognition, providing the basis for further exploration of the enzyme catalytic activities.
authors.affiliation: Department of Chemistry, University of Warwick, Library Road, Coventry, CV4 7AL, UK.
authors.forename: James S
authors.initials: JS
authors.lastname: Parascandolo
authors.affiliation: Department of Chemistry, University of Warwick, Library Road, Coventry, CV4 7AL, UK.
authors.forename: Judith
authors.initials: J
authors.lastname: Havemann
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK.
authors.forename: Helen K
authors.initials: HK
authors.lastname: Potter
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK.|Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge, CB2 1GA, UK.
authors.forename: Fanglu
authors.initials: F
authors.lastname: Huang
authors.affiliation: Department of Chemistry, University of Warwick, Library Road, Coventry, CV4 7AL, UK.
authors.forename: Elena
authors.initials: E
authors.lastname: Riva
authors.affiliation: Department of Chemistry, University of Warwick, Library Road, Coventry, CV4 7AL, UK.|School of Biosciences, The University of Birmingham, Birmingham, B15 2TT, UK.
authors.forename: Jack
authors.initials: J
authors.lastname: Connolly
authors.affiliation: Department of Chemistry, University of Warwick, Library Road, Coventry, CV4 7AL, UK.
authors.forename: Ina
authors.initials: I
authors.lastname: Wilkening
authors.affiliation: Department of Chemistry, University of Warwick, Library Road, Coventry, CV4 7AL, UK.
authors.forename: Lijiang
authors.initials: L
authors.lastname: Song
authors.affiliation: Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge, CB2 1GA, UK.
authors.forename: Peter F
authors.initials: PF
authors.lastname: Leadlay
authors.affiliation: Department of Chemistry, University of Warwick, Library Road, Coventry, CV4 7AL, UK. m.tosin@warwick.ac.uk.
authors.forename: Manuela
authors.initials: M
authors.lastname: Tosin
mesh_terms: D002851:Chromatography, High Pressure Liquid; D015335:Molecular Probes; D012459:Salicylates / Q000378:metabolism*
chemical_list: D015335:Molecular Probes; D012459:Salicylates; C005236:6-methylsalicylic acid
keywords: chemical probes; dehydratase; iterative polyketide catalysis
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