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Comparative analysis of celesticetin and lincomycin biosynthetic gene clusters
Koběrská, Markéta
Comparative analysis of celesticetin and lincomycin biosynthetic gene clusters Markéta Koběrská The introductory part of the thesis presents isolation and sequencing of lincomycin gene cluster from type strain Streptomyces lincolnensis ATCC 25466. Two relatively extensive sequence changes and several hundred point mutations were identified if compared with the previously published sequence of the lincomycin industrial strain Streptomyces lincolnensis 78-11. Analysis of the cluster flanking regions revealed its localization within the genome of S. lincolnensis ATCC 25466. The cluster-bearing cosmid was integrated into the chromosome of lincomycin non-producing strains Streptomyces coelicolor CH 999 and Streptomyces coelicolor M 145. The modified strains heterologously produced lincomycin, but the level dropped to approximately 1-3% of the production in S. lincolnensis ATCC 25466. The exact sequence of lincomycin gene cluster from the type strain allowed isolation and sequence analysis of the gene cluster of structurally related celesticetin. The analysis revealed 24 putative genes, 18 of them homologous with the genes participating in lincomycin biosynthesis. Four celesticetin specific genes are encoding enzymes involved in the salicylate biosynthesis and attachment, one is coding for celesticetin...
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Comparative analysis of celesticetin and lincomycin biosynthetic gene clusters
Koběrská, Markéta
Comparative analysis of celesticetin and lincomycin biosynthetic gene clusters Markéta Koběrská The introductory part of the thesis presents isolation and sequencing of lincomycin gene cluster from type strain Streptomyces lincolnensis ATCC 25466. Two relatively extensive sequence changes and several hundred point mutations were identified if compared with the previously published sequence of the lincomycin industrial strain Streptomyces lincolnensis 78-11. Analysis of the cluster flanking regions revealed its localization within the genome of S. lincolnensis ATCC 25466. The cluster-bearing cosmid was integrated into the chromosome of lincomycin non-producing strains Streptomyces coelicolor CH 999 and Streptomyces coelicolor M 145. The modified strains heterologously produced lincomycin, but the level dropped to approximately 1-3% of the production in S. lincolnensis ATCC 25466. The exact sequence of lincomycin gene cluster from the type strain allowed isolation and sequence analysis of the gene cluster of structurally related celesticetin. The analysis revealed 24 putative genes, 18 of them homologous with the genes participating in lincomycin biosynthesis. Four celesticetin specific genes are encoding enzymes involved in the salicylate biosynthesis and attachment, one is coding for celesticetin...
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Comparative analysis of celesticetin and lincomycin biosynthetic gene clusters
Koběrská, Markéta ; Janata, Jiří (advisor) ; Lichá, Irena (referee) ; Kormanec, Ján (referee)
Comparative analysis of celesticetin and lincomycin biosynthetic gene clusters Markéta Koběrská PhD thesis 2010 The introductory part of the thesis presents isolation and sequencing of lincomycin gene cluster from type strain Streptomyces lincolnensis ATCC 25466. Two relatively extensive sequence changes and several hundred point mutations were identified if compared with the previously published sequence of the lincomycin industrial strain Streptomyces lincolnensis 78-11. Analysis of the cluster flanking regions revealed its localization within the genome of S. lincolnensis ATCC 25466. The cluster-bearing cosmid was integrated into the chromosome of lincomycin non-producing strains Streptomyces coelicolor CH 999 and Streptomyces coelicolor M 145. The modified strains heterologously produced lincomycin, but the level dropped to approximately 1-3% of the production in S. lincolnensis ATCC 25466. The exact sequence of lincomycin gene cluster from the type strain allowed isolation and sequence analysis of the gene cluster of structurally related celesticetin. The analysis revealed 24 putative genes, 18 of them homologous with the genes participating in lincomycin biosynthesis. Four celesticetin specific genes are encoding enzymes involved in the salicylate biosynthesis and attachment, one is coding for...
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Analýza genového clusteru anthramycinových a lincosamidových antibiotik
Jelínková, Markéta ; Koběrská, Markéta ; Čermák, Lukáš ; Kopecký, Jan ; Janata, Jiří ; Spížek, Jaroslav
Lincomycin, produced by Streptomyces lincolnensis, is important, clinically used antibiotic. Its gene cluster consists of twenty-seven putative open reading frames with biosynthetic or regulatory functions and three resistance genes. Comparison of the chemical structure of the lincomycin and celesticetin indicates, that in the biosynthesis of celesticetin the whole propylproline branch is absent and proline is substrate of the penultimate step of biosynthesis, the condensation reaction. Second part of biosynthetic pathway including conversion of L-tyrosine to L-proline should be involved in the biosynthesis of functionally different anthramycin antibiotics. Thus it appears, that genetic information on lincomycin and anthramycin biosynthesis must also share common elements (genes), both biosynthetic and regulatory. The organization of transcription units was found. The analysis of the lincomycin biosynthetic gene transcripts in various cultivation stages revealed the genes with putative regulatory functions which are transcribed sooner. A set of probes based on the sequence of lincomycin biosynthetic cluster was used to search for analogous genes in celesticetin producer Streptomyces caelestis and three producers of anthramycin antibiotics: Streptomyces refuineus, Streptomyces albus and Streptosporangium sibiricum. Hybridization experiments proved presence of several analogues of genes coding for sugar moiety biosynthesis in S. caelestis and analogues of some genes coding for anthramycin antibiotics in different anthramycin producers. A rough restriction map of a part of celesticetin gene cluster presumably responsible for sugar moiety biosynthesis shows the gene arrangement to be very similar to that of lincomycin biosynthetic cluster.
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