Abstract
A general procedure for manipulating protoplasts of three Streptomyces rimosus strains was developed. More than 50% regeneration efficiency was obtained by optimizing the osmotic stabilizer concentrations and modifying the plating procedure. Preparation and regeneration of protoplasts were studied by both phase-contrast and electron microscopy. After cell wall degradation with lysozyme, protoplasts about 1,000 to 1,500 nm in diameter appeared. The reversion process exhibited normal and aberrant regeneration of protoplasts to hyphae and to spherical cells, respectively. Spherical cells contained no α, ε-ll-diaminopimelic acid and were colorless or red after Gram staining. They showed consistent stability during at least five subsequent subcultivations. However, the omission of glycine from the precultivation medium reduced the unusual process of regeneration almost completely. After normal protoplast regeneration, the production of oxytetracycline by single isolates was not affected.
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- Baltz R. H. Genetic recombination in Streptomyces fradiae by protoplast fusion and cell regeneration. J Gen Microbiol. 1978 Jul;107(1):93–102. doi: 10.1099/00221287-107-1-93. [DOI] [PubMed] [Google Scholar]
- Baltz R. H., Matsushima P. Protoplast fusion in Streptomyces: conditions for efficient genetic recombination and cell regeneration. J Gen Microbiol. 1981 Nov;127(1):137–146. doi: 10.1099/00221287-127-1-137. [DOI] [PubMed] [Google Scholar]
- Baudler E., Gumpert J. Isolation of a protoplast-type L-form from Streptomyces hygroscopicus. Z Allg Mikrobiol. 1979;19(5):363–365. doi: 10.1002/jobm.3630190509. [DOI] [PubMed] [Google Scholar]
- Gabor M. H., Hotchkiss R. D. Parameters governing bacterial regeneration and genetic recombination after fusion of Bacillus subtilis protoplasts. J Bacteriol. 1979 Mar;137(3):1346–1353. doi: 10.1128/jb.137.3.1346-1353.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hadlaczky G., Fodor K., Alföldi L. Morphological study of the reversion to bacillary form, of Bacillus megaterium protoplasts. J Bacteriol. 1976 Mar;125(3):1172–1179. doi: 10.1128/jb.125.3.1172-1179.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hopwood D. A., Chater K. F. Fresh approaches to antibiotic production. Philos Trans R Soc Lond B Biol Sci. 1980 Aug 11;290(1040):313–328. doi: 10.1098/rstb.1980.0097. [DOI] [PubMed] [Google Scholar]
- Hopwood D. A. Genetic studies with bacterial protoplasts. Annu Rev Microbiol. 1981;35:237–272. doi: 10.1146/annurev.mi.35.100181.001321. [DOI] [PubMed] [Google Scholar]
- Hopwood D. A., Wright H. M., Bibb M. J., Cohen S. N. Genetic recombination through protoplast fusion in Streptomyces. Nature. 1977 Jul 14;268(5616):171–174. doi: 10.1038/268171a0. [DOI] [PubMed] [Google Scholar]
- Okanishi M., Suzuki K., Umezawa H. Formation and reversion of Streptomycete protoplasts: cultural condition and morphological study. J Gen Microbiol. 1974 Feb;80(2):389–400. doi: 10.1099/00221287-80-2-389. [DOI] [PubMed] [Google Scholar]
- Vesligaj M., Filipović M., Pigac J., Hranueli D. Isolation of Streptomyces rimosus Mutants with Reduced Actinophage Susceptibility. Appl Environ Microbiol. 1981 Apr;41(4):986–991. doi: 10.1128/aem.41.4.986-991.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]