Fatty acyl incorporation in the biosynthesis of WAP-8294A, a group of potent anti-MRSA cyclic lipodepsipeptides

Haotong Chen, Andrew S. Olson, Wei Su, Patrick H. Dussault, Liangcheng Du

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

WAP-8294A is a family of at least 20 cyclic lipodepsipeptides exhibiting potent anti-MRSA activity. These compounds differ mainly in the hydroxylated fatty acyl chain; WAP-8294A2, the most potent member of the family that reached clinical trials, is based on (R)-3-hydroxy-7-methyloctanoic acid. It is unclear how the acyl group is incorporated because no acyl-CoA ligase (ACL) gene is present in the WAP-8294A gene cluster in Lysobacter enzymogenes OH11. Here, we identified seven putative ACL genes in the OH11 genome and showed that the yield of WAP-8294A2 was impacted by multiple ACL genes with the ACL6 gene having the most significant effect. We then investigated several (R)-3-hydroxy fatty acids and their acyl SNAC (N-acetylcysteamine) thioesters as substrates for the ACLs. Feeding (R)-3-hydroxy-7-methyloctanoate-SNAC to the ACL6 gene deletion mutant restored the production of WAP-8294A2. Finally, we heterologously expressed the seven ACL genes in E. coli and purified six of the proteins. While these enzymes exhibit a varied level of activity in vitro, ACL6 showed the highest catalytic efficiency in converting (R)-3-hydroxy-7-methyloctanoic acid to its CoA thioester when incubated with coenzyme A and ATP. These results provided both in vivo and in vitro evidence to support the fact that ACL6 is the main player for fatty acyl activation and incorporation in WAP-8294A2 biosynthesis. The results also suggest that the molecular basis for the acyl chain diversity in the WAP-8294A family is the presence of functionally overlapping ACLs.

Original languageEnglish (US)
Pages (from-to)105753-105759
Number of pages7
JournalRSC Advances
Volume5
Issue number128
DOIs
StatePublished - Jan 1 2015

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Biosynthesis
Genes
Acyl Coenzyme A
Ligases
Coenzyme A
Coenzymes
Acids
Adenosinetriphosphate
Fatty acids
Escherichia coli
Fatty Acids
Enzymes
Adenosine Triphosphate
Chemical activation
Proteins
lotilibcin
Substrates

ASJC Scopus subject areas

  • Chemistry(all)
  • Chemical Engineering(all)

Cite this

Fatty acyl incorporation in the biosynthesis of WAP-8294A, a group of potent anti-MRSA cyclic lipodepsipeptides. / Chen, Haotong; Olson, Andrew S.; Su, Wei; Dussault, Patrick H.; Du, Liangcheng.

In: RSC Advances, Vol. 5, No. 128, 01.01.2015, p. 105753-105759.

Research output: Contribution to journalArticle

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