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Abstract:

In 1937 Imshenetski and Solntseva isolated a new species of cellulose-degrading myxobacteria, which they called . 16S rRNA gene sequencing of nine isolates of and their comparison with as the reference strain proved a close phylogenetic relationship (evolutionary distance, less than 3% on the nucleotide level). The use of single antibiotics or better combinations of several antibiotics that act on different targets may be helpful, because species usually turn out to be multiresistant. Cells of the suborder “” on the one hand and “” and “” on the other hand can easily be distinguished because they differ in cell morphology, as can be detected using phase-contrast microscopy. Strains of , characterized by their intense blood-red color, and the very common strains are the only myxobacteria which degrade crystalline cellulose and can use it as the sole carbon source. At the Gesellschaft für Biotechnologische Forschung (GBF) an isolation and screening program was initiated in the late 1970s to evaluate the potential of the different genera of myxobacteria as producers of secondary metabolites. A gene product, which was previously found only in eukaryotes, has also been identified in strains. As discussed in this chapter, the fascinating microorganisms of the genus attract more and more attention, because they undergo a complex life cycle, possess the largest bacterial genomes known to date, and show a high potential as producers of biotechnologically important natural products.

Citation: Gerth K, Perlova O, Müller R. 2008. 19 , p 329-348. In Whitworth D (ed), Myxobacteria. ASM Press, Washington, DC. doi: 10.1128/9781555815677.ch19

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Reverse Transcriptase PCR
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High-Performance Liquid Chromatography
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16s rRNA Sequencing
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Figures

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Figure 1

Raster electron microscopic pictures of fruiting . (a) Vegetative swarm colony. (b) Sporangioles on the agar surface, some of which are broken. (c) Myxospores of . The surface structure is the result of drying. Pictures by K. Gerth and H. Lünsdorf.

Citation: Gerth K, Perlova O, Müller R. 2008. 19 , p 329-348. In Whitworth D (ed), Myxobacteria. ASM Press, Washington, DC. doi: 10.1128/9781555815677.ch19
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Image of Figure 2
Figure 2

Dependence of the generation time of strains on incubation temperature. So ce26 is a mesothermophilic isolate. An increase of the temperature from 30 to 40°C results in an increase of the generation time from 11 to 19 h. GT-46 and GT-41 are moderately thermophilic strains. The generation time decreases with an increase in temperature. At 42°C the temperature optimum is reached with a generation time of 6.5 h.

Citation: Gerth K, Perlova O, Müller R. 2008. 19 , p 329-348. In Whitworth D (ed), Myxobacteria. ASM Press, Washington, DC. doi: 10.1128/9781555815677.ch19
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Figure 3

Myxobacterial producers of novel secondary metabolites. With 47% of total production, strains are the most outstanding producers of novel metabolites.

Citation: Gerth K, Perlova O, Müller R. 2008. 19 , p 329-348. In Whitworth D (ed), Myxobacteria. ASM Press, Washington, DC. doi: 10.1128/9781555815677.ch19
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Figure 4

Frequency of some selected metabolites derived from strains. The data are given as numbers of producer strains from 1,700 screened isolates. From Gerth et al. (2003) with the permission of Elsevier, B.V.

Citation: Gerth K, Perlova O, Müller R. 2008. 19 , p 329-348. In Whitworth D (ed), Myxobacteria. ASM Press, Washington, DC. doi: 10.1128/9781555815677.ch19
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Image of Figure 5
Figure 5

A survey of “novel” metabolites from . Typical linear and macrocyclic polyketides are presented. Some of them are likely to be biosynthesized by combinations of peptide synthetases and polyketide synthetases, e.g., eliamid. Socein is one of the rare polypeptides active against fungi and yeasts.

Citation: Gerth K, Perlova O, Müller R. 2008. 19 , p 329-348. In Whitworth D (ed), Myxobacteria. ASM Press, Washington, DC. doi: 10.1128/9781555815677.ch19
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Figure 6

(a) Structures of myxobacterial secondary metabolites with a benzoic acid moiety. (b) Benzoyl-CoA biosynthesis in So ce26 (soraphen producer). Mutants E4 and E5 are nonproducer mutants. Mutant E4 excretes traces of cinnamic acid and high concentrations of phenyl propionic acid into the culture supernantant. Mutant E5 recovers the ability of soraphen production in the presence of these compounds. From Gerth et al. (2003) with the permission of Elsevier, B.V.

Citation: Gerth K, Perlova O, Müller R. 2008. 19 , p 329-348. In Whitworth D (ed), Myxobacteria. ASM Press, Washington, DC. doi: 10.1128/9781555815677.ch19
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Figure 7

Transposon mutagenesis in . (a) -based transposon. IR, inverted repeats, P, promoter of the gene; Ω, transcription terminator of the hygromycin resistance gene (Hyg); oriR6Kγ, conditional origin of replication. (b) Transposon region when integrated into the chromosome. (c) Transposon recovery, consisting of ligation of chromosomal DNA from mutants after restriction with an enzyme which does not cut inside the transposed element (e.g., MluI). Using Primer 1 and Primer 2 the flanking chromosomal regions can be sequenced from the recovered plasmid. (d) Analysis of mutants, using a bioassay (e.g., comparison of nonproducers of chivosazol obtained by transposon mutagenesis with the wild type, which shows an inhibition zone on the indicator plate) (Kopp et al., 2004).

Citation: Gerth K, Perlova O, Müller R. 2008. 19 , p 329-348. In Whitworth D (ed), Myxobacteria. ASM Press, Washington, DC. doi: 10.1128/9781555815677.ch19
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Figurer 8

Identification of the chivosazol biosynthetic gene cluster by inactivation of PKS genes using plasmid integration by homologous recombination. (a) Inactivation plasmid containing the selection marker Hyg and a homologous region obtained by PCR using degenerate PKS primers. (b) Biosynthetic gene cluster; localization of mutations is marked. (c) HPLC chromatogram of culture extracts of So ce56 wild type (wt) and chivosazol-negative mutants (Mutant1 and Mutant2). (d) Bioassay for chivosazol production using .

Citation: Gerth K, Perlova O, Müller R. 2008. 19 , p 329-348. In Whitworth D (ed), Myxobacteria. ASM Press, Washington, DC. doi: 10.1128/9781555815677.ch19
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