Release of autolytic enzyme from Streptococcus, faecium cell walls by treatment with dilute alkali

Cornett, J B; Johnson, Clifton; Shockman, Gérald D.

doi:10.1128/jb.138.3.699-704.1979

Cited by 16 publications

(3 citation statements)

References 14 publications

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“…Susceptibility to lysis was, however, influenced by an increase of pH above neutrality, since the cells which were grown to the end of exponential phase did not lyse, if the pH was above 8 (Fig, 6). It is feasible that the alkaline conditions inactivated the autolysins [14]. If the exponential-phase organisms were harvested and then diluted back to the supernatant fluid with and without pH adjustment to 6.8, the cells grew only for a while, before they started to lyse.…”

Section: Methodsmentioning

confidence: 99%

Autolysis of Streptococcus thermophilus

Sandholm¹

1981

FEMS Microbiology Letters

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Section: Methodsmentioning

confidence: 99%

Autolysis of Streptococcus thermophilus

Sandholm¹

1981

FEMS Microbiology Letters

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“…Lithium chloride has been previously used to extract cell wall-associated proteins in L. helveticus and in particular the S-layerforming protein (29). In addition, this chemical agent has been successfully used to extract autolysins of various species such as Clostridium perfringens (55), Bacillus subtilis (8,42), and Streptococcus faecium (9,38).…”

Section: Vol 61 1995mentioning

confidence: 99%

Zymogram and Preliminary Characterization of Lactobacillus helveticus Autolysins

Valence¹,

Lortal²

1995

Appl Environ Microbiol

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The autolysins of Lactobacillus helveticus ISLC5 were detected and partially characterized by renaturing sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis with substrate-containing gels (zymogram). By using lyophilized Micrococcus luteus cells or heated whole cells of L. helveticus ISLC5 (0.2% [wt/vol]) as a substrate, several lytic activities were detected in the whole-cell SDS extract of strain ISLC5 (i) one activity at 42.4 kDa, which was named autolysin A, and (ii) six other activities having very similar molecular weights (29.1, 29.6, 30, 30.8, 31.7, and 32.8 kDa), which were named autolysins B (B1 through B6, respectively). As regards the temporal distribution of the enzymes, autolysins A and B were detected in the cells harvested from the beginning of the exponential growth phase. Autolysin A appeared to be associated only with viable cells, whereas the autolysins B remained associated with the cell envelope several days after the complete loss of culture viability. When SDS-treated walls of L. helveticus ISLC5 were used as a substrate, a supplementary lytic activity appeared at 37.5 kDa; it was considered a peptidoglycan hydrolase, since it was not able to induce lysis of whole-cell substrate. The autolysins of 30 other strains of L. helveticus from various geographical origins were also analyzed by zymogram; all the activity profiles obtained were similar to that of strain ISLC5 in terms of the number of lytic bands and their apparent molecular weights. Only the relative intensities of the lytic bands corresponding to autolysins A and B were variable depending on the strains. This observation suggested that autolysins are highly conserved enzymes. A concentrated crude lysate of the virulent bacteriophage 832-B1 infecting L. helveticus was also analyzed by zymogram; one lytic activity with an apparent molecular weight of 31.7 kDa, very close to the weights of the autolysins B, was observed. Finally, the autolysins of L. helveticus ISLC5 were successfully extracted from whole cells by using a 1 M lithium chloride solution; they were partially purified by precipitation, selective resolubilization, and gel filtration chromatography, which led to a 20-fold increase in specific activity.

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“…Identifying proteins has never been more sensitive, less laborious, and easier due to the availability of improved analytical technologies and biochemical kits [ 2 , 12 ]. However, sensitive analytical detection may be compromised by contaminating proteins/peptides introduced as artifacts during experimental processes, as has been observed when using lithium chloride, tris-buffered urea, and trypsin surface-shaving surface protein extraction methods whereby these methods allow cell leakage of cytoplasmic proteins [ 13 , 14 , 15 , 16 , 17 ]. The Matrix Assisted Laser Ionization Time-of-Flight Mass Spectrometer (MALDI-TOF MS) analyzes individually-isolated proteins.…”

Section: Introductionmentioning

confidence: 99%

Comparison of Surface Proteomes of Adherence Variants of Listeria Monocytogenes Using LC-MS/MS for Identification of Potential Surface Adhesins

2016

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The ability of Listeria monocytogenes to adhere and form biofilms leads to persistence in food processing plants and food-associated listeriosis. The role of specific surface proteins as adhesins to attach Listeria cells to various contact surfaces has not been well characterized to date. In prior research comparing different methods for surface protein extraction, the Ghost urea method revealed cleaner protein content as verified by the least cytoplasmic protein detected in surface extracts using LC-MS/MS. The same technique was utilized to extract and detect surface proteins among two surface-adherent phenotypic strains of L. monocytogenes (i.e., strongly and weakly adherent). Of 640 total proteins detected among planktonic and sessile cells, 21 protein members were exclusively detected in the sessile cells. Relative LC-MS/MS detection and quantification of surface-extracted proteins from the planktonic weakly adherent (CW35) and strongly adherent strains (99-38) were examined by protein mass normalization of proteins. We found that L. monocytogenes 99-38 exhibited a total of 22 surface proteins that were over-expressed: 11 proteins were detected in surface extracts of both sessile and planktonic 99-38 that were ≥5-fold over-expressed while another 11 proteins were detected only in planktonic 99-38 cells that were ≥10-fold over-expressed. Our results suggest that these protein members are worthy of further investigation for their involvement as surface adhesins.

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Release of autolytic enzyme from Streptococcus, faecium cell walls by treatment with dilute alkali

Cited by 16 publications

References 14 publications

Autolysis of Streptococcus thermophilus

Autolysis of Streptococcus thermophilus

Zymogram and Preliminary Characterization of Lactobacillus helveticus Autolysins

Comparison of Surface Proteomes of Adherence Variants of Listeria Monocytogenes Using LC-MS/MS for Identification of Potential Surface Adhesins

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