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Engineered biosynthesis of nonribosomal lipopeptides with modified fatty acid side chains.

Powell, A, Borg, M, Amir-Heidari, B, Neary, JM, Thirlway, J, Wilkinson, B, Smith, CP and Micklefield, J (2007) Engineered biosynthesis of nonribosomal lipopeptides with modified fatty acid side chains. J Am Chem Soc, 129 (49). pp. 15182-15191.

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Abstract

The biological properties of the calcium-dependent antibiotics (CDAs), daptomycin and related nonribosomal lipopeptides, depend to a large extent on the nature of the N-terminal fatty acid moiety. It is suggested that the chain length of the unusually short (C6) 2,3-epoxyhexanoyl fatty acid moiety of CDA is determined by the specificity of the KAS-II enzyme encoded by fabF3 in the CDA biosynthetic gene cluster. Indeed, deletion of the downstream gene hxcO results in three new lipopeptides, all of which possess hexanoyl side chains (hCDAs). This confirms that HxcO functions as a hexanoyl-CoA or -ACP oxidase. The absence of additional CDA products with longer fatty acid groups further suggests that the CDA lipid chain is biosynthesized on a single ACP and is then transferred directly from this ACP to the first CDA peptide synthetase (CdaPS1). Interestingly, the hexanoyl-containing CDAs retain antibiotic activity. To further modulate the biological properties of CDA by introducing alternative fatty acid groups, a mutasynthesis approach was developed. This involved mutating the key active site Ser residue of the CdaPS1, module 1 PCP domain to Ala, which prevents subsequent phosphopantetheinylation. In the absence of the natural module 1 PCP tethered intermediate, it is possible to effect incorporation of different N-acyl-L-serinyl N-acetylcysteamine (NAC) thioester analogues, leading to CDA products with pentanoyl as well as hexanoyl side chains.

Item Type: Article
Authors :
NameEmailORCID
Powell, AUNSPECIFIEDUNSPECIFIED
Borg, MUNSPECIFIEDUNSPECIFIED
Amir-Heidari, BUNSPECIFIEDUNSPECIFIED
Neary, JMUNSPECIFIEDUNSPECIFIED
Thirlway, JUNSPECIFIEDUNSPECIFIED
Wilkinson, BUNSPECIFIEDUNSPECIFIED
Smith, CPc.p.smith@surrey.ac.ukUNSPECIFIED
Micklefield, JUNSPECIFIEDUNSPECIFIED
Date : 12 December 2007
Identification Number : https://doi.org/10.1021/ja074331o
Uncontrolled Keywords : 3-Oxoacyl-(Acyl-Carrier-Protein) Synthase, Anti-Bacterial Agents, Daptomycin, Escherichia coli, Fatty Acids, Lipoproteins, Mutagenesis, Site-Directed, Protein Engineering, Streptomyces coelicolor, Synechocystis
Related URLs :
Depositing User : Symplectic Elements
Date Deposited : 17 May 2017 09:02
Last Modified : 17 May 2017 09:02
URI: http://epubs.surrey.ac.uk/id/eprint/821968

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