Heterologous Expression, Purification, and Characterization of Type II Polyketide Synthase Acyl Carrier Proteins

Grayson S Hamrick; Casey H Londergan; Louise K Charkoudian

doi:10.1007/978-1-0716-2273-5_13

Heterologous Expression, Purification, and Characterization of Type II Polyketide Synthase Acyl Carrier Proteins

Methods Mol Biol. 2022:2489:239-267. doi: 10.1007/978-1-0716-2273-5_13.

Authors

Grayson S Hamrick¹, Casey H Londergan¹, Louise K Charkoudian²

Affiliations

¹ Department of Chemistry, Haverford College, Haverford, PA, USA.
² Department of Chemistry, Haverford College, Haverford, PA, USA. lcharkou@haverford.edu.

Abstract

The enzymes that comprise type II polyketide synthases (PKSs) are powerful biocatalysts that, once well-understood and strategically applied, could enable cost-effective and sustainable access to a range of pharmaceutically relevant molecules. Progress toward this goal hinges on gaining ample access to materials for in vitro characterizations and structural analysis of the components of these synthases. A central component of PKSs is the acyl carrier protein (ACP), which serves as a hub during the biosynthesis of type II polyketides. Herein, we share methods for accessing type II PKS ACPs via heterologous expression in E. coli . We also share how the installation of reactive and site-specific spectroscopic probes can be leveraged to study the conformational dynamics and interactions of type II PKS ACPs.

Keywords: Acyl carrier protein; Heterologous expression; Polyaromatic polyketide; Polyketide synthase; Type II polyketide.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, N.I.H., Extramural

MeSH terms

Acyl Carrier Protein* / genetics
Acyl Carrier Protein* / metabolism
Escherichia coli / metabolism
Polyketide Synthases* / genetics

Substances

Acyl Carrier Protein
Polyketide Synthases

Grants and funding

R15 GM120704/GM/NIGMS NIH HHS/United States