The mucopeptide turnover in the cell walls of growing cultures ofBacillus megaterium KM

Chaloupka, J.; Křečková, P.; Řfhová, L.

doi:10.1007/bf02172250

Cited by 29 publications

(15 citation statements)

References 6 publications

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“…Cell wall turnover has also been demonstrated in a variety of other gram-positive organisms including Streptococcus aureus (31), Lac-tobacillus acidophilus (3,11), and B. megaterium (4)(5)(6)21), each of which shows somewhat different kinetic characteristics than the turnover observed with B. subtilis.…”

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confidence: 99%

Relation between cell wall turnover and cell growth in Bacillus subtilis

Glaser¹,

Lindsay²

1977

J Bacteriol

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The kinetics of cell wall turnover in Bacillus subtilis have been examined in detail. After pulse labeling of the peptidoglycan with N-acetylglucosamine, the newly formed peptidoglycan is stable for approximately three-quarters of a generation and is then degraded by a process that follows first-order kinetics. Deprivation ofan auxotroph ofamino acids required for protein synthesis results in a cessation of turnover. If a period of amino acid starvation occurs during the lag phase of turnover, then the initiation of turnover is delayed for a period of time equivalent to the starvation period. During amino acid starvation, new cell wall peptidoglycan is synthesized and added to preexisting cell wall. This peptidoglycan after resumption of growth is also subject to degradation (turnover). It is suggested that cell wall turnover is dependent on cell growth and elongation. Several possible control mechanisms for cell wall autolytic enzymes are discussed in light of these observations.The cell wall peptidoglycan of many grampositive microorganisms turns over during logarithmic growth (3-6, 11, 12, 21, 22, 26, 27, 31). A number of observations suggest that peptidoglycan turnover is not required for normal cell growth (13); however, the phenomenon is of interest because it may provide clues to the mode of cell wall growth and assembly and some insight into the regulation of cell wall autolytic enzymes.Previous work with Bacillus subtilis has shown that after pulse labeling of the peptidoglycan, there is a lag period, equivalent in time to 0.5 to 1.0 generations, during which the newly synthesized peptidoglycan is immune from turnover. After this lag period, the peptidoglycan is degraded by a process that follows first-order kinetics, and the products ofpeptidoglycan turnover (glycan strands and peptide bridges) appear in the growth medium (21). If any fraction of the peptidoglycan in this organism is resistant to turnover, it must represent less than 5% of the total cell wall (12,21). Although the loss of radioactivity from the cell wall is about 50% per generation, it has been pointed out by Pooley (26,27) that since there is a lag between the time at which peptidoglycan is synthesized and the time at which it is degraded, the loss ofpeptidoglycan from the cell is only 10 to 20% of the mass of the cell wall per generation.Cell wall turnover has also been demonstrated in a variety of other gram-positive organisms including Streptococcus aureus (31), Lac-tobacillus acidophilus (3,11), and B. megaterium (4-6, 21), each of which shows somewhat different kinetic characteristics than the turnover observed with B. subtilis.In this communication we have examined some of the kinetics of cell wall turnover in B. subtilis, and its relation to cell growth. The results appear to place important restrictions on any model of cell wall expansion in this organism. MATERIALS AND METHODSThe bacterial strains used were either the Marburg strain of B. subtilis (ATCC 6051) or a derivative of this strain, B. subtilis B42, kindly made availab...

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confidence: 99%

Relation between cell wall turnover and cell growth in Bacillus subtilis

Glaser¹,

Lindsay²

1977

J Bacteriol

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“…The first report describing the bacterial cell wall turnover was published for the Gram-positive bacterium Bacillus megaterium more than 50 years ago (23). Later on, pulse-chased experiments demonstrated with radioactively labeled cell wall precursors that all studied Grampositive bacteria carry out a cell wall turnover as well as Gram-negative bacteria (14,34,81,107).…”

Section: Turnover Of Peptidoglycanmentioning

confidence: 99%

The Gram-Positive Bacterial Cell Wall

Rohde

2019

Microbiol Spectr

119

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The chapter about the Gram-positive bacterial cell wall gives a brief historical background on the discovery of Gram-positive cell walls and their constituents and microscopic methods applied for studying the Gram-positive cell envelope. Followed by the description of the different chemical building blocks of peptidoglycan and the biosynthesis of the peptidoglycan layers and high turnover of peptidoglycan during bacterial growth. Lipoteichoic acids and wall teichoic acids are highlighted as major components of the cell wall. Characterization of capsules and the formation of extracellular vesicles by Gram-positive bacteria close the section on cell envelopes which have a high impact on bacterial pathogenesis. In addition, the specialized complex and unusual cell wall of mycobacteria is introduced thereafter. Next a short back view is given on the development of electron microscopic examinations for studying bacterial cell walls. Different electron microscopic techniques and methods applied to examine bacterial cell envelopes are discussed in the view that most of the illustrated methods should be available in a well-equipped life sciences orientated electron microscopic laboratory. In addition, newly developed and mostly well-established cryo-methods like high-pressure freezing and freeze-substitution (HPF-FS) and cryo-sections of hydrated vitrified bacteria (CEMOVIS, Cryo-electron microscopy of vitreous sections) are described. At last, modern cryo-methods like cryo-electron tomography (CET) and cryo-FIB-SEM milling (focus ion beam-scanning electron microscopy) are introduced which are available only in specialized institutions, but at present represent the best available methods and techniques to study Gram-positive cell walls under close-to-nature conditions in great detail and at high resolution.

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“…PG degradation research has shown that a large portion of degradation intermediates are reutilized during bacterial cell growth. This cell wall turnover phenomenon was discovered in the early 1960s byŘíhová L. and coworkers, and the activity of cell wall degradation has since been discovered in a variety of Gram-positive and Gram-negative bacteria, including E. coli (Chaloupka, Křečková,Řfhová, 1962;Chaloupka et al, 2008;Doyle, Chaloupka, & Vinter, 1988;Park & Uehara, 2008). Using radiolabeled PG from E. coli, Goodell and Schwarz calculated the percentage of PG that is lost during each generation and identified intermediate molecules from the cell medium (Goodell, 1985;Goodell & Schwarz, 1985).…”

Section: Background Informationmentioning

confidence: 99%

Metabolic Incorporation of N‐Acetyl Muramic Acid Probes into Bacterial Peptidoglycan

DeMeester

Liang

Zhou

et al. 2019

CP Chemical Biology

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Bacterial cells utilize small carbohydrate building blocks to construct peptidoglycan (PG), a highly conserved mesh‐like polymer that serves as a protective coat for the cell. PG production has long been a target for antibiotics, and its breakdown is a source for human immune recognition. A key component of bacterial PG, N‐acetyl muramic acid (NAM), is a vital element in many synthetically derived immunostimulatory compounds. However, the exact molecular details of these structures and how they are generated remain unknown due to a lack of chemical probes surrounding the NAM core. A robust synthetic strategy to generate bioorthogonally tagged NAM carbohydrate units is implemented. These molecules serve as precursors for PG biosynthesis and recycling. Escherichia coli cells are metabolically engineered to incorporate the bioorthogonal NAM probes into their PG network. The probes are subsequently modified using copper‐catalyzed azide‐alkyne cycloaddition to install fluorophores directly into the bacterial PG, as confirmed by super‐resolution microscopy and high‐resolution mass spectrometry. Here, synthetic notes for key elements of this process to generate the sugar probes as well as streamlined user‐friendly metabolic labeling strategies for both microbiology and immunological applications are described. © 2019 by John Wiley & Sons, Inc. Basic Protocol 1: Synthesis of peracetylated 2‐azido glucosamine Basic Protocol 2: Synthesis of 2‐azido and 2‐alkyne NAM Basic Protocol 3: Synthesis of 3‐azido NAM methyl ester Basic Protocol 4: Incorporation of NAM probes into bacterial peptidoglycan Basic Protocol 5: Confirmation of bacterial cell wall remodeling by mass spectrometry

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The mucopeptide turnover in the cell walls of growing cultures ofBacillus megaterium KM

Cited by 29 publications

References 6 publications

Relation between cell wall turnover and cell growth in Bacillus subtilis

Relation between cell wall turnover and cell growth in Bacillus subtilis

The Gram-Positive Bacterial Cell Wall

Metabolic Incorporation of N‐Acetyl Muramic Acid Probes into Bacterial Peptidoglycan

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