Versatility of Acyl-Acyl Carrier Protein Synthetases

Beld, Joris; Finzel, Kara; Burkart, Michael D.

doi:10.1016/j.chembiol.2014.08.015

Title: Versatility of Acyl-Acyl Carrier Protein Synthetases

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Abstract

The acyl carrier protein (ACP) requires posttranslational modification with a 4'-phosphopantetheine arm for activity, and this thiol-terminated modification carries cargo between enzymes in ACP-dependent metabolic pathways. In this paper, we show that acyl-ACP synthetases (AasSs) from different organisms are able to load even, odd, and unnatural fatty acids onto E. coli ACP in vitro. Vibrio harveyi AasS not only shows promiscuity for the acid substrate, but also is active upon various alternate carrier proteins. AasS activity also extends to functional activation in living organisms. We show that exogenously supplied carboxylic acids are loaded onto ACP and extended by the E. coli fatty acid synthase, including unnatural fatty acid analogs. These analogs are further integrated into cellular lipids. Finally, in vitro characterization of four different adenylate-forming enzymes allowed us to disambiguate CoA-ligases and AasSs, and further in vivo studies show the potential for functional application in other organisms.

Authors:: Beld, Joris; Finzel, Kara; Burkart, Michael D.

Publication Date:: Wed Oct 01 00:00:00 EDT 2014

Research Org.:: Univ. of California, San Diego, CA (United States)

Sponsoring Org.:: USDOE Office of Energy Efficiency and Renewable Energy (EERE), Sustainable Transportation Office. Bioenergy Technologies Office

OSTI Identifier:: 1224136

Alternate Identifier(s):: OSTI ID: 1222379; OSTI ID: 1344393

Grant/Contract Number:: EE0003373

Resource Type:: Published Article

Journal Name:: Chemistry & Biology

Additional Journal Information:: Journal Name: Chemistry & Biology Journal Volume: 21 Journal Issue: 10; Journal ID: ISSN 1074-5521

Publisher:: Elsevier

Country of Publication:: United Kingdom

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES

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                    Beld, Joris, Finzel, Kara, and Burkart, Michael D. Versatility of Acyl-Acyl Carrier Protein Synthetases.  United Kingdom: N. p., 2014. 
Web.  doi:10.1016/j.chembiol.2014.08.015.

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                    Beld, Joris, Finzel, Kara, & Burkart, Michael D. Versatility of Acyl-Acyl Carrier Protein Synthetases.  United Kingdom.  https://doi.org/10.1016/j.chembiol.2014.08.015

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                    Beld, Joris, Finzel, Kara, and Burkart, Michael D. Wed .  
"Versatility of Acyl-Acyl Carrier Protein Synthetases".  United Kingdom.  https://doi.org/10.1016/j.chembiol.2014.08.015.

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@article{osti_1224136,

  title        = {Versatility of Acyl-Acyl Carrier Protein Synthetases},

  author       = {Beld, Joris and Finzel, Kara and Burkart, Michael D.},

  abstractNote = {The acyl carrier protein (ACP) requires posttranslational modification with a 4'-phosphopantetheine arm for activity, and this thiol-terminated modification carries cargo between enzymes in ACP-dependent metabolic pathways. In this paper, we show that acyl-ACP synthetases (AasSs) from different organisms are able to load even, odd, and unnatural fatty acids onto E. coli ACP in vitro. Vibrio harveyi AasS not only shows promiscuity for the acid substrate, but also is active upon various alternate carrier proteins. AasS activity also extends to functional activation in living organisms. We show that exogenously supplied carboxylic acids are loaded onto ACP and extended by the E. coli fatty acid synthase, including unnatural fatty acid analogs. These analogs are further integrated into cellular lipids. Finally, in vitro characterization of four different adenylate-forming enzymes allowed us to disambiguate CoA-ligases and AasSs, and further in vivo studies show the potential for functional application in other organisms.},

  doi          = {10.1016/j.chembiol.2014.08.015},

  journal      = {Chemistry & Biology},

  number       = 10,

  volume       = 21,

  place        = {United Kingdom},

  year         = {Wed Oct 01 00:00:00 EDT 2014},

  month        = {Wed Oct 01 00:00:00 EDT 2014}

}

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