@article{mbs:/content/journal/micro/10.1099/00221287-133-9-2595, author = "REYSSET, GILLES and HUBERT, JACQUES and PODVIN, LAURENCE and SEBALD, MADELEINE", title = "Protoplast Formation and Regeneration of Clostridium acetobutylicum Strain N1-4080", journal= "Microbiology", year = "1987", volume = "133", number = "9", pages = "2595-2600", doi = "https://doi.org/10.1099/00221287-133-9-2595", url = "https://www.microbiologyresearch.org/content/journal/micro/10.1099/00221287-133-9-2595", publisher = "Microbiology Society", issn = "1465-2080", type = "Journal Article", abstract = "SUMMARY: A method for the production of protoplasts of Clostridium acetobutylicum strain N1-4080 utilizing lysozyme and penicillin G was optimized to yield about 100% protoplasts. Treatment of 109 cells left fewer than 102 that were viable and osmo-resistant. With a mutant strain deficient in autolytic activity, about 1% of the protoplasts regenerated walled cells, whereas most gave rise to stable L-form colonies. Addition of sodium polyanethole sulphonate to the medium improved the regeneration frequency by at least a factor of 10, probably by somehow inhibiting the residual autolytic activity of the strain.", }