Genetic engineering and heterologous expression of the disorazol biosynthetic gene cluster via Red/ET recombineering.
Cast your vote
You can rate an item by clicking the amount of stars they wish to award to this item.
When enough users have cast their vote on this item, the average rating will also be shown.
Your vote was cast
Thank you for your feedback
Thank you for your feedback
MetadataShow full item record
AbstractDisorazol, a macrocyclic polykitide produced by the myxobacterium Sorangium cellulosum So ce12 and it is reported to have potential cytotoxic activity towards several cancer cell lines, including multi-drug resistant cells. The disorazol biosynthetic gene cluster (dis) from Sorangium cellulosum (So ce12) was identified by transposon mutagenesis and cloned in a bacterial artificial chromosome (BAC) library. The 58-kb dis core gene cluster was reconstituted from BACs via Red/ET recombineering and expressed in Myxococcus xanthus DK1622. For the first time ever, a myxobacterial trans-AT polyketide synthase has been expressed heterologously in this study. Expression in M. xanthus allowed us to optimize the yield of several biosynthetic products using promoter engineering. The insertion of an artificial synthetic promoter upstream of the disD gene encoding a discrete acyl transferase (AT), together with an oxidoreductase (Or), resulted in 7-fold increase in disorazol production. The successful reconstitution and expression of the genetic sequences encoding for these promising cytotoxic compounds will allow combinatorial biosynthesis to generate novel disorazol derivatives for further bioactivity evaluation.
CitationGenetic engineering and heterologous expression of the disorazol biosynthetic gene cluster via Red/ET recombineering. 2016, 6:21066 Sci Rep
AffiliationHelmholtz-Institute for Pharmaceutical Research Saarland (HIPS),Saarland 9 University, 66123 Saarbrücken, Germany.
The following license files are associated with this item:
- Production of the tubulin destabilizer disorazol in Sorangium cellulosum: biosynthetic machinery and regulatory genes.
- Authors: Kopp M, Irschik H, Pradella S, Müller R
- Issue date: 2005 Jul
- Heterologous expression and genetic engineering of the tubulysin biosynthetic gene cluster using Red/ET recombineering and inactivation mutagenesis.
- Authors: Chai Y, Shan S, Weissman KJ, Hu S, Zhang Y, Müller R
- Issue date: 2012 Mar 23
- Heterologous expression of epothilone biosynthetic genes in Myxococcus xanthus.
- Authors: Julien B, Shah S
- Issue date: 2002 Sep
- Reconstitution of the myxothiazol biosynthetic gene cluster by Red/ET recombination and heterologous expression in Myxococcus xanthus.
- Authors: Perlova O, Fu J, Kuhlmann S, Krug D, Stewart AF, Zhang Y, Müller R
- Issue date: 2006 Dec
- Modular construction of a functional artificial epothilone polyketide pathway.
- Authors: Osswald C, Zipf G, Schmidt G, Maier J, Bernauer HS, Müller R, Wenzel SC
- Issue date: 2014 Oct 17