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Dive into the research topics where Marta Fedoryshyn is active.

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Featured researches published by Marta Fedoryshyn.


Gene | 2008

Marker removal from actinomycetes genome using Flp recombinase

Marta Fedoryshyn; Lutz Petzke; Elisabeth Welle; Andreas Bechthold; Andriy Luzhetskyy

We report here a system for the functional expression of the Flp recombinase in several actinomycetes: Streptomyces coelicolor, S. lividans, and Saccharotrix espanaensis. We have constructed a synthetic gene encoding the Flp recombinase with a GC content of 60.6% optimized for expression in high-GC bacteria. Using the synthetic flp(a) gene, we have removed an apramycin resistance gene flanked by FRT sites from the chromosome of actinomycetes with an efficiency of 40%. Sequencing the region of chromosome showed that excision of the apramycin cassette by Flp recombinase was specific.


Applied Microbiology and Biotechnology | 2008

Functional expression of the Cre recombinase in actinomycetes

Marta Fedoryshyn; Elisabeth Welle; Andreas Bechthold; Andriy Luzhetskyy

Site-specific recombinases revolutionized “in vivo” genetic engineering because they can catalyze precise excisions, integrations, inversions, or translocations of DNA between their distinct recognition target sites. We have constructed a synthetic gene encoding Cre recombinase with the GC content 67.7% optimized for expression in high-GC bacteria and demonstrated this gene to be functional in Streptomyces lividans. Using the synthetic cre(a) gene, we have removed an apramycin resistance gene flanked by loxP sites from the chromosome of S. lividans with 100% efficiency. Sequencing of the chromosomal DNA part showed that excision of the apramycin cassette by Cre recombinase was specific.


ChemBioChem | 2005

Generation of novel landomycins M and O through targeted gene disruption.

Andriy Luzhetskyy; Lili Zhu; Miranda Gibson; Marta Fedoryshyn; Clemens Dürr; Carsten Hofmann; Dirk Hoffmeister; Bohdan Ostash; Cynthia Mattingly; Val R. Adams; Victor Fedorenko; Jürgen Rohr; Andreas Bechthold

Two genes from Streptomyces cyanogenous S136 that encode the reductase LanZ4 and the hydroxylase LanZ5, which are involved in landomycin A biosynthesis, were characterized by targeted gene inactivation. Analyses of the corresponding mutants as well as complementation experiments have allowed us to show that LanZ4 and LanZ5 are responsible for the unique C‐11‐hydroxylation that occurs during landomycin biosynthesis. Compounds accumulated by the lanZ4/Z5 mutants are the previously described landomycin F and the new landomycins M and O.


ChemBioChem | 2005

LanGT2 Catalyzes the First Glycosylation Step during landomycin A biosynthesis.

Andriy Luzhetskyy; Takaaki Taguchi; Marta Fedoryshyn; Clemens Dürr; Sven-Eric Wohlert; Volodymyr Novikov; Andreas Bechthold

The glycosyltransferase LanGT2 is involved in the biosynthesis of the hexasaccharide side chain of the angucyclic antibiotic landomycin A. Its function was elucidated by targeted gene inactivation of lanGT2. The main metabolite of the obtained mutant was identified as tetrangulol (4), the progenitor of the landomycin aglycon (7). The lack of the sugar side chain indicates that LanGT2 catalyzes the priming glycosyl transfer in the hexasaccharide biosynthesis: the attachment of a D‐olivose to O‐8 of the polyketide backbone. Heterologous expression of urdGT2 from S. fradiae Tü2717 in this mutant resulted in the production of a novel C‐glycosylated angucycline (6).


Russian Journal of Genetics | 2006

IncP Plasmids are Most Effective in Mediating Conjugation between Escherichia coli and Streptomycetes

Andriy Luzhetskyy; Marta Fedoryshyn; Oleksandr Gromyko; Bohdan Ostash; Y. Rebets; Andreas Bechthold; V. Fedorenko

Mobilizable shuttle plasmids containing the origin of transfer (oriT) region of plasmid F (IncFI), ColIb-P9 (IncI1), and RP4/RP1 (IncPα) were constructed to test the ability of the cognate conjugation system to mediate gene transfer from Escherichia coli to Streptomyces. The conjugative system of the IncPα plasmids was shown to be most effective in conjugative transfer, giving peak values of (2.7 ± 0.2) × 10−2S. lividans TK24 exconjugants per recipient cell. To assess whether the mating-pair formation system or the DNA-processing apparatus of the IncPα plasmids is crucial in conjugative transfer, an assay with an IncQ-based mobilizable plasmid (RSF1010) specifying its own DNA-processing system was developed. Only the IncPα plasmid mobilized the construct to S. lividans indicating that the mating-pair formation system is primarly responsible for the promiscuous transfer of the plasmids between E. coli and Streptomyces. Dynamic of conjugative transfer from E. coli to S. lividans was investigated and exconjugants starting from the first hour of mating were obtained.


Chemistry & Biology | 2009

Engineering a Function into a Glycosyltransferase

Christine Krauth; Marta Fedoryshyn; Christian Schleberger; Andriy Luzhetskyy; Andreas Bechthold

As glycosyltransferases found in nature often show distinct substrate specificity, glycosyltransferase engineering is an important research field. In this work, we were able to introduce an activity into a glycosyltransferase involved in natural product (landomycin E) biosynthesis. This was achieved by recognizing hot spot amino acids in glycosyltransferases which are strongly involved in determining substrate specificity.


ChemBioChem | 2004

Function of lanGT3, a Glycosyltransferase Gene Involved in Landomycin A Biosynthesis

Andriy Luzhetskyy; Tao Liu; Marta Fedoryshyn; Bohdan Ostash; V. Fedorenko; Jürgen Rohr; Andreas Bechthold

The glycosyltransferase gene lanGT3, involved in the biosynthesis of the angucyclic antibiotic landomycin A, has been characterised by targeted gene deletion. A lanGT3 mutant was shown to produce landomycin E, which consists of a trisaccharide side chain attached to the polyketide moiety. Expression of lanGT3 in the mutant restored landomycin A production. Our results indicate that LanGT3 is responsible for the transfer of the fourth sugar during landomycin A biosynthesis.


Chemistry & Biology | 2005

Iteratively Acting Glycosyltransferases Involved in the Hexasaccharide Biosynthesis of Landomycin A

Andriy Luzhetskyy; Marta Fedoryshyn; Clemens Dürr; Takaaki Taguchi; Volodymyr Novikov; Andreas Bechthold


Journal of Biotechnology | 2007

Surprising production of a new urdamycin derivative by S. fradiae ΔurdQ/R

Marta Fedoryshyn; Mohammad Nur-e-Alam; Lili Zhu; Andriy Luzhetskyy; J. Rohr; A. Bechthold


Archive | 2002

A GENE CLONING SYSTEM FOR STREPTOMYCES CYANOGENUS S136

Andriy Luzhetskyy; Marta Fedoryshyn; Dirk Hoffmeister; Andreas Bechthold; V. Fedorenko; Ivan Franko; Albert-Ludwigs-University Freiburg; Pharmazeutische Biologie

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Lutz Petzke

University of Freiburg

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