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. 1986 Oct;52(4):612–616. doi: 10.1128/aem.52.4.612-616.1986

Protoplast formation and regeneration of dehydrodivanillin-degrading strains of Fusobacterium varium and Enterococcus faecium.

W Chen, K Ohmiya, S Shimizu
PMCID: PMC239085  PMID: 3777921

Abstract

Two strains of rumen anaerobes isolated from dehydrodivanillin-degrading cultures were identified as Fusobacterium varium and Enterococcus faecium. These organisms degraded dehydrodivanillin synergistically to 5-carboxymethylvanillin and vanillic acid. Specific conditions for protoplast formation and cell wall regeneration for both bacteria were determined, under strictly anaerobic conditions, to be as follows. The cell wall of each bacterium in yeast extract medium was loosened by adding penicillin G during early log-phase growth. The cell wall of F. varium was lysed by lysozyme (1 mg/ml) in glycerol (0.2 M)-phosphate buffer (0.05 M; pH 7.0). The addition of NaCl (0.08 M) with lysozyme was necessary for lysis of E. faecium in this solution. Almost all cells were converted to protoplasts after 2 h of incubation at 37 degrees C. Regeneration of both protoplasts was 20 to 30% on an agar-containing yeast extract medium.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allcock E. R., Reid S. J., Jones D. T., Woods D. R. Clostridium acetobutylicum Protoplast Formation and Regeneration. Appl Environ Microbiol. 1982 Mar;43(3):719–721. doi: 10.1128/aem.43.3.719-721.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chen W., Ohmiya K., Shimizu S., Kawakami H. Degradation of dehydrodivanillin by anaerobic bacteria from cow rumen fluid. Appl Environ Microbiol. 1985 Jan;49(1):211–216. doi: 10.1128/aem.49.1.211-216.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Fodor K., Alföldi L. Fusion of protoplasts of Bacillus megaterium. Proc Natl Acad Sci U S A. 1976 Jun;73(6):2147–2150. doi: 10.1073/pnas.73.6.2147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gold M. H., Cheng T. M., Alic M. Formation, Fusion, and Regeneration of Protoplasts from Wild-Type and Auxotrophic Strains of the White Rot Basidiomycete Phanerochaete chrysosporium. Appl Environ Microbiol. 1983 Jul;46(1):260–263. doi: 10.1128/aem.46.1.260-263.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Knowlton S., Ferchak J. D., Alexander J. K. Protoplast Regeneration in Clostridium tertium: Isolation of Derivatives with High-Frequency Regeneration. Appl Environ Microbiol. 1984 Dec;48(6):1246–1247. doi: 10.1128/aem.48.6.1246-1247.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Lee-Wickner L. J., Chassy B. M. Production and Regeneration of Lactobacillus casei Protoplasts. Appl Environ Microbiol. 1984 Nov;48(5):994–1000. doi: 10.1128/aem.48.5.994-1000.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Metcalf R. H., Deibel R. H. Differential lytic response of enterococci associated with addition order of lysozyme and anions. J Bacteriol. 1969 Sep;99(3):674–680. doi: 10.1128/jb.99.3.674-680.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Minton N. P., Morris J. G. Regeneration of protoplasts of Clostridium pasteurianum ATCC 6013. J Bacteriol. 1983 Jul;155(1):432–434. doi: 10.1128/jb.155.1.432-434.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ohmiya Y., Angevine L. S., Mehendale H. M. Effect of drug-induced phospholipidosis on pulmonary disposition of pneumophilic drugs. Drug Metab Dispos. 1983 Jan-Feb;11(1):25–30. [PubMed] [Google Scholar]
  11. 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]
  12. Pettey T. M., Crawford D. L. Enhancement of Lignin Degradation in Streptomyces spp. by Protoplast Fusion. Appl Environ Microbiol. 1984 Feb;47(2):439–440. doi: 10.1128/aem.47.2.439-440.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Power J. B., Cummins S. E., Cocking E. C. Fusion of isolated plant protoplasts. Nature. 1970 Mar 14;225(5237):1016–1018. doi: 10.1038/2251016a0. [DOI] [PubMed] [Google Scholar]
  14. Smith M. D. Transformation and fusion of Streptococcus faecalis protoplasts. J Bacteriol. 1985 Apr;162(1):92–97. doi: 10.1128/jb.162.1.92-97.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Weiss R. L. Protoplast formation in Escherichia coli. J Bacteriol. 1976 Nov;128(2):668–670. doi: 10.1128/jb.128.2.668-670.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]

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