Untersuchungen zur Biosynthese von Fumitremorgin-Typs in Aspergillus fumigatus und Neosartorya fischeri sowie Produktion von gewünschten Substanzen mittels chemoenzymatischer Synthese
Final Report Abstract
The two genes AFUA_8G00150 and AFUA_8G00160 in the neighbourhood of the fumitremorgin gene cluster were suggested in our proposal to be responsible for the formation of fumitremorgin A and sprirotryprostatins, respectively. Using recombinant proteins, no evidence could be provided for this hypothesis. Instead, we have demonstrated that the formation of fumitremorgin A from verruculogen was surprisingly catalysed by the prenyltransferase FtmpT3 outside of the fumitremorgin/verruculogen cluster. The coding gene ftmPT3 was found even at a different chromosome in the genome of Neosartorya fischeri as the mentioned gene cluster. Identification of FtmPT3 as the verruculogen prenyltransferase completed the reaction steps in the biosynthetic pathway from L-tryptophan and L-proline to fumitremorgin A in fungi. Meanwhile, this work demonstrated also the power of combination of different disciplines. The formation of sprirotryprostatins was catalysed by the fumitremorgin C hydroxylase FtmP450-3, as demonstrated recently by Tsunematsu et al. This enzyme was originally proposed to be responsible only for the conversion of fumitremorgin C to 12, 13-dihydroxyfumitremorgin C. As planned, we have successfully produced 14 tryprostatin analogues with significantly increased cytotoxicity by using FtmPT1 as biocatalysts. These results prompted us to get cyclic dipeptides with prenyl moieties at other positions of the indole ring and to test their cytotoxicity, which is now under investigation. Production of fumitremorgin C analogues by using FtmPT1 and FtmP450-2 and testing their biological activity are still under investigation. This is also true for the investigation of the mechanism of the FtmOx1 reaction.
Publications
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(2011). Genome mining and biosynthesis of fumitremorgin-type alkaloids in ascomycetes. Journal of Antibiotics, 64:45-49
Shu-Ming Li
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(2012). Breaking the regioselectivity of indole prenyltransferases: identification of regular C3-prenylated hexahydropyrrolo[2,3-b]indoles as side products of the regular C2-prenyltransferase FtmPT1. Organic and Biomolecular Chemistry 10: 9262-9270
Beate Wollinsky, Lena Ludwig, Xiulan Xie and Shu-Ming Li
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(2012). Identification of the verruculogen prenyltransferase FtmPT3 by a combination of chemical, bioinformatic and biochemical approaches. ChemBioChem 13: 2583-2592
Kathrin Mundt, Beate Wollinsky, Han-Li Ruan, Tianjiao Zhu and Shu-Ming Li
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(2012). Prenylation at the indole ring leads to a significant increase of cytotoxicity of tryptophan-containing cyclic dipeptides. Bioorganic and Medicinal Chemistry Letters 22: 3866–3869
Beate Wollinsky, Lena Ludwig, Alexandra Hamacher, Xia Yu, Matthias Kassack and Shu-Ming Li
