Identification of Lipstatin-Producing Ability in Streptomyces virginiae CBS 314.55 Using Dereplication Approach

Gordan Sladič1, Matilda Urukalo2, Matjaž Kirn1, Urška Lešnik3, Vasilka Magdevska3,
Neda Benički1, Mitja Pelko1, Aleš Gasparič1, Peter Raspor2, Tomaž Polak2, Štefan Fujs3, Paul A. Hoskisson4* and Hrvoje Petković2,3*

Krka d.d., Novo mesto, Šmarješka cesta 6, SI-8501 Novo Mesto, Slovenia

2University of Ljubljana, Biotechnical Faculty, Jamnikarjeva 101, SI-1000 Ljubljana, Slovenia
3Acies Bio d.o.o., Tehnološki park 21, SI-1000 Ljubljana, Slovenia
4Strathclyde Institute of Pharmacy and Biomedical Science, University of Strathclyde, Hamnett Wing,161 Cathedral Street, Glasgow, G4 0RE, UK

Article history

Received April 5, 2013

Accepted February 25, 2014

Key words

lipstatin, Streptomyces virginiae, dereplication, lipase inhibitor


species are prolific producers of bioactive metabolites, such as β-lactone
-containing lipstatin produced by Streptomyces toxytricini, an intermediate used in semi-synthetic process for production of anti-obesity drug orlistat. Understanding the distribution of identical or structurally similar molecules produced by a taxonomic group is of particular importance when trying to isolate novel biologically active compounds or strains producing known metabolites of medical importance with potentially improved properties. Until now, only two independent isolates of S. toxytricini species have been known to be producers of lipstatin. According to the current taxonomic criteria, S. toxytricini belongs to Streptomyces lavendulae phenotypic cluster. Taxonomy-based dereplication approach coupled with in vitro assay was applied to screen the S. lavendulae phenotypic cluster for production of lipstatin-like lipase inhibitors using synthetic p-nitrophenol derivatives of C4 and C16 lipids. Screening the available strains from public collections belonging to S. lavendulae phenotypic cluster, high lipase inhibitory activity was identified in the Streptomyces virginiae CBS 314.55 culture supernatants. HPLC and LC-MS/MS confirmed lipstatin production by a new Streptomyces species for the first time. We have demonstrated that the new lipstatin-producing strain S. virginiae morphologically and physiologically differs from S. toxytricini substantially; however, the production capacity of the newly identified lipstatin-producing species S. virginiae is comparable to S. toxytricini. We have thus demonstrated the effectiveness of a simple and affordable dereplication approach for identification of potentially novel and useful industrial strains available in public culture collections.



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