Journal article
Strategic Acyl Carrier Protein Engineering Enables Functional Type II Polyketide Synthase Reconstitution In Vitro
ACS chemical biology, v 20(1), pp 197-207
17 Jan 2025
PMID: 39745931
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Microbial polyketides represent a structurally diverse class of secondary metabolites with medicinally relevant properties. Aromatic polyketides are produced by type II polyketide synthase (PKS) systems, each minimally composed of a ketosynthase-chain length factor (KS-CLF) and a phosphopantetheinylated acyl carrier protein (
-ACP). Although type II PKSs are found throughout the bacterial kingdom, and despite their importance to strategic bioengineering, type II PKSs have not been well-studied
. In cases where the KS-CLF can be accessed via
heterologous expression, often the cognate ACPs are not activatable by the broad specificity
surfactin-producing phosphopantetheinyl transferase (PPTase) Sfp and, conversely, in systems where the ACP can be activated by Sfp, the corresponding KS-CLF is typically not readily obtained. Here, we report the high-yield heterologous expression of both cyanobacterial
sp. PCC 7428 minimal type II PKS (gloPKS) components in
, which allowed us to study this minimal type II PKS
. Initially, neither the cognate PPTase nor Sfp converted gloACP to its active
state. However, by examining sequence differences between Sfp-compatible and -incompatible ACPs, we identified two conserved residues in gloACP that, when mutated, enabled high-yield phosphopantetheinylation of gloACP by Sfp. Using analogous mutations, other previously Sfp-incompatible type II PKS ACPs from different bacterial phyla were also rendered activatable by Sfp. This demonstrates the generalizability of our approach and breaks down a longstanding barrier to type II PKS studies and the exploration of complex biosynthetic pathways.
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Details
- Title
- Strategic Acyl Carrier Protein Engineering Enables Functional Type II Polyketide Synthase Reconstitution In Vitro
- Creators
- Kevin Li - Haverford CollegeYae In Cho - Haverford CollegeMai Anh Tran - Friedrich Schiller University JenaChristoph Wiedemann - Friedrich Schiller University JenaShuaibing Zhang - Leibniz-Institut für Naturstoff-Forschung und Infektionsbiologie e. V. - Hans-Knöll-Institut (HKI)Rebecca S Koweek - Haverford CollegeNgọc Khánh Hoàng - Haverford CollegeGrayson S Hamrick - Haverford CollegeMargaret A Bowen - Haverford CollegeBashkim Kokona - Drexel UniversityPierre Stallforth - Friedrich Schiller University JenaJoris Beld - Drexel University, Microbiology and ImmunologyUte A Hellmich - Friedrich Schiller University JenaLouise K Charkoudian - Haverford College
- Publication Details
- ACS chemical biology, v 20(1), pp 197-207
- Publisher
- ACS Publications
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Microbiology and Immunology
- Web of Science ID
- WOS:001388635900001
- Scopus ID
- 2-s2.0-85215254884
- Other Identifier
- 991022016114104721
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- Collaboration types
- Domestic collaboration
- International collaboration
- Web of Science research areas
- Biochemistry & Molecular Biology