Abstract
A novel microbial-screening procedure was developed for separate detection of 1,4-butanediol, ethylene glycol, and adipic acid, three commercially important oxychemicals potentially derivable from bacterial omega-oxidation of n-butanol, ethanol, and hexanoic acid, respectively. The screening method involved postproduction addition of one of several specific Pseudomonas strains which produce a soluble fluorescent pigment during growth on the product of interest. A mutation and selection procedure was developed for isolation of specific strains with phenotypes for growth and pigment production on the desired product (e.g., 1,4-butanediol), but not on its bioconversion substrate (e.g., n-butanol), common by-products (e.g., n-butyrate), or product isomers. Pigment production was growth associated and required cultivation of the screening strains under limiting Fe3+ concentrations. The pigments resembled well-characterized, iron-chelating siderophores produced by other fluorescent pseudomonads. The sensitivity of the assay for product accumulation was enhanced by (i) conducting the screening in microtiter dishes to permit examination of individual isolates of putative producers and to control product diffusion, (ii) using a wavelength cutoff filter to reduce background source light, and (iii) using adapted screening strains which grew at lower (0.3 mM) concentrations of test compounds. The potential utility of the method for detecting a variety of oxidative catabolic products is discussed.
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Clarke P. H. The metabolic versatility of pseudomonads. Antonie Van Leeuwenhoek. 1982 May;48(2):105–130. doi: 10.1007/BF00405197. [DOI] [PubMed] [Google Scholar]
- Leisinger T., Margraff R. Secondary metabolites of the fluorescent pseudomonads. Microbiol Rev. 1979 Sep;43(3):422–442. doi: 10.1128/mr.43.3.422-442.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ornston L. N., Ornston M. K., Chou G. Isolation of spontaneous mutant strains of Pseudomonas putida. Biochem Biophys Res Commun. 1969 Jul 7;36(1):179–184. doi: 10.1016/0006-291x(69)90666-4. [DOI] [PubMed] [Google Scholar]
- Teintze M., Hossain M. B., Barnes C. L., Leong J., van der Helm D. Structure of ferric pseudobactin, a siderophore from a plant growth promoting Pseudomonas. Biochemistry. 1981 Oct 27;20(22):6446–6457. doi: 10.1021/bi00525a025. [DOI] [PubMed] [Google Scholar]
- Teintze M., Leong J. Structure of pseudobactin A, a second siderophore from plant growth promoting Pseudomonas B10. Biochemistry. 1981 Oct 27;20(22):6457–6462. doi: 10.1021/bi00525a026. [DOI] [PubMed] [Google Scholar]