Rational engineering acyltransferase domain of modular polyketide synthase for expanding substrate specificity

2019 ◽  
pp. 271-292 ◽  
Author(s):  
Wan Zhang ◽  
Linjun Zhou ◽  
Chunyu Li ◽  
Zixin Deng ◽  
Xudong Qu
Keyword(s):  
Substrate Specificity ◽  
Polyketide Synthase ◽  
Rational Engineering ◽  
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 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

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.

scholarly journals Substrate structure–activity relationships guide rational engineering of modular polyketide synthase ketoreductases

2016 ◽  
Vol 52 (4) ◽  
pp. 792-795 ◽  
Author(s):  
Constance B. Bailey ◽  
Marjolein E. Pasman ◽  
Adrian T. Keatinge-Clay
Keyword(s):  
Polyketide Synthase ◽  
Structure Activity Relationship ◽  
Active Enzyme ◽  
Activity Relationship ◽  
Structure Activity Relationships ◽  
Rational Engineering ◽  
Substrate Structure ◽  
Structure Activity ◽  
Modular Polyketide Synthase

Structure–activity relationship studies guided stereocontrol engineering within a modular polyketide synthase ketoreductase to yield a more active enzyme whose reactivity can be explained through the Felkin–Anh model.

scholarly journals Successes, surprises and pitfalls in modular polyketide synthase engineering: generation of ring-contracted stambomycins

2021 ◽  
Author(s):  
Li Su ◽  
Laurence Hotel ◽  
Cédric Paris ◽  
Alexander Brachmann ◽  
Jörn Piel ◽  
...  
Keyword(s):  
Polyketide Synthase ◽  
Modular Organization ◽  
Efficient Production ◽  
Type I ◽  
Wild Type ◽  
Rational Engineering ◽  
Substantial Modification ◽  
Poor Tolerance ◽  
Multiple State ◽  
Modular Polyketide Synthase

Abstract The modular organization of the type I polyketide synthases (PKSs) would seem propitious for rational engineering of desirable analogous. However, despite decades of efforts, such experiments remain largely inefficient. Here, we combined multiple, state-of-the-art approaches including modification of docking domains, use of modules of varying domain composition, alternative interdomain fusion sites, and targeted adaptation of key domain-domain interfaces, to reprogram the stambomycin PKS by deleting seven internal modules – the most substantial modification to an intact system reported to date. One such system produced the target 37-membered mini-stambomycin metabolites, a reduction in chain length of 14 carbons relative to the 51-membered parental compounds, but also substantial quantities of shunt metabolites released from the multienzyme subunit upstream of the newly-installed junction. Our data also provide evidence for an unprecedented off-loading mechanism of such stalled intermediates involving the C-terminal thioesterase domain acting on chains located four modules upstream. The yields of all metabolites were substantially reduced compared to the wild type compounds, likely reflecting the poor tolerance to the non-native substrates of the modules downstream of the introduced interfaces. Taken together, our data demonstrate that even ‘optimized’ PKS engineering strategies remain inadequate for efficient production of target polyketide derivatives, and highlight several areas for future investigation.

Bioinformatics-Guided Discovery of Citrulline 4-Hydroxylase from GE81112 Biosynthesis and Rational Engineering of Its Substrate Specificity

2019 ◽  
Author(s):  
Christian Zwick ◽  
Max Sosa ◽  
Hans Renata
Keyword(s):  
Substrate Specificity ◽  
Guided Discovery ◽  
Rational Engineering

We functionally characterize a nonheme dioxygenase from GE81112 biosynthesis and identify it as a citrulline hydroxylase. A bioinformatics guided engineering was performed to alter the substrate specificity of the enzyme.

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
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