Production of Protoplasts of Escherichia Coli by Lysozyme Treatment

Zinder, Norton D.; Arndt, William F.

doi:10.1073/pnas.42.9.586

Cited by 71 publications

(33 citation statements)

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“…The dual features of the cell-wall structure of Gram-negative bacteria suggested above is compatible with observations of the loss of cell-wall rigidity and of ' protoplast ' formation by lysozyme treatment of Escherichia coli a t high pH values (Zinder & Arndt, 1956) or with 'Versene' (Mahler & Fraser, 1956) and the production of ' protoplasts ' of Escherichia coli on media containing penicillin (Lederberg, 1956). Recent experiments performed by Dr F. Shafa and myself have indicated that both Iysozyme action and penicillin action probably involve the same type of structure in the cell walls of certain Gram-negative bacteria.…”

Section: Bacterial Lysissupporting

confidence: 86%

The Lysis of Micro-organisms by Lysozyme and Related Enzymes

Saltón

1958

Journal of General Microbiology

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Section: Bacterial Lysissupporting

confidence: 86%

The Lysis of Micro-organisms by Lysozyme and Related Enzymes

Saltón

1958

Journal of General Microbiology

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“…However, analogous wall-less cells were generated in just this way over 40 years ago (26,34). For historical and semantic consistency, we have adopted the terminology of Birdsell and Cota-Robles and refer to these as LI spheroplasts.…”

Section: Discussionmentioning

confidence: 99%

“…Interestingly, bacteria that have lost their cell wall can sometimes resynthesize this structure and recover a wild-type shape, indicating that cells are able to generate a defined morphology without the aid of a previously completed external template (25)(26)(27)(28). What is not clear is whether the currently known shape maintenance pathways are sufficient for directing the recovery of cell shape in wall-less cells.…”

mentioning

confidence: 99%

The Rcs Stress Response and Accessory Envelope Proteins Are Required for De Novo Generation of Cell Shape in Escherichia coli

Ranjit

Young

2013

J Bacteriol

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Interactions with immune responses or exposure to certain antibiotics can remove the peptidoglycan wall of many Gram-negative bacteria. Though the spheroplasts thus created usually lyse, some may survive by resynthesizing their walls and shapes. Normally, bacterial morphology is generated by synthetic complexes directed by FtsZ and MreBCD or their homologues, but whether these classic systems can recreate morphology in the absence of a preexisting template is unknown. To address this question, we treated Escherichia coli with lysozyme to remove the peptidoglycan wall while leaving intact the inner and outer membranes and periplasm. The resulting lysozyme-induced (LI) spheroplasts recovered a rod shape after four to six generations. Recovery proceeded via a series of cell divisions that produced misshapen and branched intermediates before later progeny assumed a normal rod shape. Importantly, mutants defective in mounting the Rcs stress response and those lacking penicillin binding protein 1B (PBP1B) or LpoB could not divide or recover their cell shape but instead enlarged until they lysed. LI spheroplasts from mutants lacking the Lpp lipoprotein or PBP6 produced spherical daughter cells that did not recover a normal rod shape or that did so only after a significant delay. Thus, to regenerate normal morphology de novo, E. coli must supplement the classic FtsZ-and MreBCD-directed cell wall systems with activities that are otherwise dispensable for growth under normal laboratory conditions. The existence of these auxiliary mechanisms implies that they may be required for survival in natural environments, where bacterial walls can be damaged extensively or removed altogether.

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“…More especially, it has been found that under defined environmental conditions, lysozyme may also initiate lysis of Gram-negative cells (Repaske, 1956;Zinder & Arndt, 1956). If bacterial lysis resulting from phage action is really due to the presence of a lytic substance, formed during phage development and acting 'from inside ' on infected cells, one may wonder whether such a substance would not require the same conditions as lysozyme in order to act 'from outside ' on non-infected cells.…”

Section: The Presence Of Endolysin In the Lysates Of Induced Lysogenimentioning

confidence: 99%