scholarly journals Engineering the Substrate Specificity of a Modular Polyketide Synthase for Installation of Consecutive Non-Natural Extender Units

2019 ◽  
Vol 141 (5) ◽  
pp. 1961-1969 ◽  
Author(s):  
Edward Kalkreuter ◽  
Jared M. CroweTipton ◽  
Andrew N. Lowell ◽  
David H. Sherman ◽  
Gavin J. Williams
Keyword(s):  
Substrate Specificity ◽  
Polyketide Synthase ◽  
Modular Polyketide Synthase

Alteration of the Substrate Specificity of a Modular Polyketide Synthase Acyltransferase Domain through Site-Specific Mutations†

Biochemistry ◽  
2001 ◽  
Vol 40 (51) ◽  
pp. 15464-15470 ◽  
Author(s):  
Christopher D. Reeves ◽  
Sumati Murli ◽  
Gary W. Ashley ◽  
Misty Piagentini ◽  
C. Richard Hutchinson ◽  
...  
Keyword(s):  
Substrate Specificity ◽  
Polyketide Synthase ◽  
Site Specific ◽  
Modular Polyketide Synthase

scholarly journals Type II Thioesterase ScoT, Associated with Streptomyces coelicolor A3(2) Modular Polyketide Synthase Cpk, Hydrolyzes Acyl Residues and Has a Preference for Propionate

2008 ◽  
Vol 75 (4) ◽  
pp. 887-896 ◽  
Author(s):  
Magdalena Kotowska ◽  
Krzysztof Pawlik ◽  
Aleksandra Smulczyk-Krawczyszyn ◽  
Hubert Bartosz-Bechowski ◽  
Katarzyna Kuczek
Keyword(s):  
Substrate Specificity ◽  
Kinetic Parameters ◽  
Hybrid Systems ◽  
Active Site ◽  
Polyketide Synthase ◽  
Streptomyces Coelicolor ◽  
Polyketide Synthases ◽  
Type Ii ◽  
Starter Unit ◽  
Modular Polyketide Synthase

ABSTRACT Type II thioesterases (TE IIs) were shown to maintain the efficiency of polyketide synthases (PKSs) by removing acyl residues blocking extension modules. However, the substrate specificity and kinetic parameters of these enzymes differ, which may have significant consequences when they are included in engineered hybrid systems for the production of novel compounds. Here we show that thioesterase ScoT associated with polyketide synthase Cpk from Streptomyces coelicolor A3(2) is able to hydrolyze acetyl, propionyl, and butyryl residues, which is consistent with its editing function. This enzyme clearly prefers propionate, in contrast to the TE IIs tested previously, and this indicates that it may have a role in control of the starter unit. We also determined activities of ScoT mutants and concluded that this enzyme is an α/β hydrolase with Ser90 and His224 in its active site.

Promiscuity of a modular polyketide synthase towards natural and non-natural extender units

2013 ◽  
Vol 11 (27) ◽  
pp. 4449 ◽  
Author(s):  
Irina Koryakina ◽  
John B. McArthur ◽  
Matthew M. Draelos ◽  
Gavin J. Williams
Keyword(s):  
Polyketide Synthase ◽  
Modular Polyketide Synthase

scholarly journals Type II thioesterase ScoT is required for coelimycin production by the modular polyketide synthase Cpk of Streptomyces coelicolor A3(2).

2014 ◽  
Vol 61 (1) ◽  
Author(s):  
Magdalena Kotowska ◽  
Jarosław Ciekot ◽  
Krzysztof Pawlik
Keyword(s):  
Culture Medium ◽  
Polyketide Synthase ◽  
Streptomyces Coelicolor ◽  
Acyl Chain ◽  
No Production ◽  
Type I ◽  
Type Ii ◽  
Peptide Synthetases ◽  
Modular Polyketide Synthase ◽  
Editing Enzyme

Type II thioesterases were shown to maintain efficiency of modular type I polyketide synthases and nonribosomal peptide synthetases by removing acyl residues blocking extension modules. We found that thioesterase ScoT from Streptomyces coelicolor A3(2) is required for the production of the yellow-pigmented coelimycin by the modular polyketide synthase Cpk. No production of coelimycin was observed in cultures of scoT disruption mutant. Polyketide production was restored upon complementation with an intact copy of the scoT gene. An enzymatic assay showed that ScoT thioesterase can hydrolyse a 12-carbon acyl chain but the activity is too low to play a role in product release from the polyketide synthase. We conclude that ScoT is an editing enzyme necessary to maintain the activity of polyketide synthase Cpk. We provide a HPLC based method to measure the amount of coelimycin P2 in a culture medium.

Probing the Flexibility of an Iterative Modular Polyketide Synthase with Non-Native Substrates in Vitro

2018 ◽  
Vol 13 (8) ◽  
pp. 2261-2268 ◽  
Author(s):  
Samuel C. Curran ◽  
Andrew Hagen ◽  
Sean Poust ◽  
Leanne Jade G. Chan ◽  
Brett M. Garabedian ◽  
...  
Keyword(s):  
Polyketide Synthase ◽  
Modular Polyketide Synthase

An Evolutionary Model Encompassing Substrate Specificity and Reactivity of Type I Polyketide Synthase Thioesterases

ChemBioChem ◽  
2014 ◽  
Vol 15 (18) ◽  
pp. 2656-2661 ◽  
Author(s):  
Taylor P. A. Hari ◽  
Puneet Labana ◽  
Meaghan Boileau ◽  
Christopher N. Boddy
Keyword(s):  
Substrate Specificity ◽  
Polyketide Synthase ◽  
Evolutionary Model ◽  
Type I
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