Faculty Opinions recommendation of Comparative analysis of the substrate specificity of trans- versus cis-acyltransferases of assembly line polyketide synthases.

Polyketide natural products act as a broad range of therapeutics, including antibiotics, immunosuppressants and anti-cancer agents. This therapeutic diversity stems from the structural diversity of these small molecules, many of which are produced in an assembly line manner by modular polyketide synthases. The acyltransferase (AT) domains of these megasynthases are responsible for selection and incorporation of simple monomeric building blocks, and are thus responsible for a large amount of the resulting polyketide structural diversity. The substrate specificity of these domains is often targeted for engineering in the generation of novel, therapeutically active natural products. This review outlines recent developments that can be used in the successful engineering of these domains, including AT sequence and structural data, mechanistic insights and the production of a diverse pool of extender units. It also provides an overview of previous AT domain engineering attempts, and concludes with proposed engineering approaches that take advantage of current knowledge. These approaches may lead to successful production of biologically active ‘unnatural’ natural products.

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A comparative analysis of the kinetic mechanism and peptide substrate specificity of human and Saccharomyces cerevisiae myristoyl-CoA:protein N-myristoyltransferase

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)82159-7 ◽

1993 ◽

Vol 268 (14) ◽

pp. 9964-9971

Author(s):

W.J. Rocque ◽

C.A. McWherter ◽

D.C. Wood ◽

J.I. Gordon

Keyword(s):

Saccharomyces Cerevisiae ◽

Comparative Analysis ◽

Substrate Specificity ◽

Kinetic Mechanism ◽

Peptide Substrate

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Mixed-model assembly line sequencing heuristics for smoothing component parts usage: A comparative analysis

International Journal of Production Research ◽

10.1080/002075498192850 ◽

1998 ◽

Vol 36 (8) ◽

pp. 2209-2224 ◽

Cited By ~ 29

Author(s):

E. A. Duplaga ◽

D.J. Bragg

Keyword(s):

Comparative Analysis ◽

Assembly Line ◽

Mixed Model ◽

Mixed Model Assembly Line ◽

Mixed Model Assembly

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Computational Approach for Prediction of Domain Organization and Substrate Specificity of Modular Polyketide Synthases

Journal of Molecular Biology ◽

10.1016/s0022-2836(03)00232-8 ◽

2003 ◽

Vol 328 (2) ◽

pp. 335-363 ◽

Cited By ~ 161

Author(s):

Gitanjali Yadav ◽

Rajesh S Gokhale ◽

Debasisa Mohanty

Keyword(s):

Substrate Specificity ◽

Computational Approach ◽

Polyketide Synthases ◽

Domain Organization

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Computational Studies on the Substrate Specificity of an Acyltransferase Domain from Salinomycin Polyketide Synthase

Catalysis Science & Technology ◽

10.1039/d1cy00284h ◽

2021 ◽

Author(s):

Huining Ji ◽

Ting Shi ◽

Lei Liu ◽

Fa Zhang ◽

Wentao Tao ◽

...

Keyword(s):

Natural Products ◽

Substrate Specificity ◽

Polyketide Synthase ◽

Coenzyme A ◽

Biological Activities ◽

Polyketide Synthases ◽

Short Chain ◽

Computational Studies ◽

Chemical Structures ◽

Coa Thioesters

Polyketides are a large group of natural products with diverse chemical structures and biological activities. They are biosynthesized by modular polyketide synthases (PKSs) from coenzyme A (CoA) thioesters of short-chain...

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In Vitro Investigation of Crosstalk between Fatty Acid and Polyketide Synthases in the Andrimid Biosynthetic Assembly Line

ChemBioChem ◽

10.1002/cbic.201600410 ◽

2016 ◽

Vol 17 (22) ◽

pp. 2137-2142 ◽

Cited By ~ 9

Author(s):

Fumihiro Ishikawa ◽

Hiroyasu Sugimoto ◽

Hideaki Kakeya

Keyword(s):

Fatty Acid ◽

Assembly Line ◽

Polyketide Synthases ◽

In Vitro Investigation

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An in vitro platform for engineering and harnessing modular polyketide synthases

Nature Communications ◽

10.1038/s41467-019-13811-0 ◽

2020 ◽

Vol 11 (1) ◽

Cited By ~ 7

Author(s):

Takeshi Miyazawa ◽

Melissa Hirsch ◽

Zhicheng Zhang ◽

Adrian T. Keatinge-Clay

Keyword(s):

Assembly Line ◽

Polyketide Synthases ◽

Assembly Lines ◽

Dihydroxybenzoic Acid ◽

Coa Ligase ◽

Malonyl Coa ◽

Aromatic Product ◽

Order Of Magnitude ◽

Line Engineering

AbstractTo harness the synthetic power of modular polyketide synthases (PKSs), many aspects of their biochemistry must be elucidated. A robust platform to study these megadalton assembly lines has not yet been described. Here, we in vitro reconstitute the venemycin PKS, a short assembly line that generates an aromatic product. Incubating its polypeptides, VemG and VemH, with 3,5-dihydroxybenzoic acid, ATP, malonate, coenzyme A, and the malonyl-CoA ligase MatB, venemycin production can be monitored by HPLC and NMR. Multi-milligram quantities of venemycin are isolable from dialysis-based reactors without chromatography, and the enzymes can be recycled. Assembly line engineering is performed using pikromycin modules, with synthases designed using the updated module boundaries outperforming those using the traditional module boundaries by over an order of magnitude. Using combinations of VemG, VemH, and their engineered derivatives, as well as the alternate starter unit 3-hydroxybenzoic acid, a combinatorial library of six polyketide products is readily accessed.

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