Effects of wash treatments on the ultrastructure and lysozyme penetrability of the outer membrane of various marine and two terrestrial gram-negative bacteria

Laddaga, Richard A.; MacLeod, Robert A.

doi:10.1139/m82-047

Cited by 19 publications

(9 citation statements)

References 22 publications

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“…Isolation of OM by extraction of cell envelopes in 10 mM Tris HCI (pH 7.4) containing 0.5% (wt/vol) of Sarkosyl which preferentially solubilized the CM of Escherichia coli (16) also failed because the detergent solubilized both the OM and the CM. However, a modification of a method for fractionation of cell envelopes of the marine bacterium Alteromonas haloplanktis (19,40) which involved sequentially washing cells in 0.5 M NaCl and 25% (wt/vol) sucrose was successful. from cells at this stage of growth, and when the cells were washed with a mineral solution corresponding in composition to the medium, the OM remained intact.…”

Section: Resultsmentioning

confidence: 99%

“…DISCUSSION In this study the OM from glucose-and cellulose-grown cells of F. succinogenes was isolated by using a NaCl and sucrose wash procedure previously developed for the isolation of cell envelope fractions from the marine bacterium Alteromonas haloplanktis (19). Successful application of the method to F. succinogenes cells is based on the observation that the bacterium has a comparatively high salt requirement for optimal growth (4), a characteristic in common with marine bacteria (40). The release of membrane fragments into the wash solutions was coincident with the loss of OM from F. succinogenes cells.…”

Section: Resultsmentioning

confidence: 99%

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Separation of outer and cytoplasmic membranes of Fibrobacter succinogenes and membrane and glycogen granule locations of glycanases and cellobiase

Gong

Forsberg

1993

J Bacteriol

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The outer membrane (OM) of Fibrobacter succinogenes was isolated by a combination of salt, sucrose, and water washes from whole cells grown on either glucose or cellulose. The cytoplasmic membrane (CM) was isolated from OM-depleted cells after disruption with a French press. The OM and membrane vesicles isolated from the extracellular culture fluid of cellulose-grown cells had a higher density, much lower succinate dehydrogenase activity, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profiles different from those of the CM. The OM from both glucose-and cellulose-grown cells and the extracellular membrane vesicles from cellulose-grown cultures exhibited higher endoglucanase, xylanase, and acetylesterase activities than the CM and other cell fractions. Endoglucanase 2 was absent from the isolated OM fractions of glucose-and cellulose-grown cells and from the extracellular membrane vesicles of cellulose-grown cells but was present in the CM and intracellular glycogen granule fractions, while endoglucanase 3 was enriched in the OM. Cellobiosidase was located primarily in the periplasm as previously reported, while cellobiase was mainly present in the glycogen granule fraction of glucose-grown cells and in a nongranular glycogen and CM complex in cellulose-grown cells. The cellobiase was not eluted from glycogen granules by cellobiose, maltose, and maltotriose nor from either the granules or the cell membranes by nondenaturing detergents but was eluted from both glycogen granules and cell membranes by high concentrations of salts. The eluted cellobiase rebound almost quantitatively when diluted and mixed with purified glycogen granules but exhibited a low affinity for Avicel cellulose. Thus, we have documented a method for isolation of OM from F. succinogenes, identified the OM origin of the extracellular membrane vesicles, and located glycanases and cellobiase in membrane and glycogen fractions.Fibrobacter succinogenes S85 is one of the major cellulolytic bacteria in the rumen of animals receiving low-quality forage rations (44). It extensively degrades cellulose after attachment to the insoluble substrate (39), which indicates that the cellulase enzymes are functioning at the cell surface during cellulose digestion. Nevertheless, the mechanism by which the bacterium degrades cellulose is still unknown (18,44). To elucidate the physiological mechanism of cellulose degradation by the bacterium, three endoglucanases, endoglucanase 1 (EGI), EG2 (46), and EG3 (49), a chloride-stimulated cellobiosidase (31), and a cellodextrinase (28, 29) were purified and characterized. EG I was released from cells during growth, and

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Separation of outer and cytoplasmic membranes of Fibrobacter succinogenes and membrane and glycogen granule locations of glycanases and cellobiase

Gong

Forsberg

1993

J Bacteriol

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“…The bottles were placed in a 25°C shaking (140 rpm) incubator for 3 days, and during this time the cells reduced Se(IV) to Se(0), as indicated by the appearance of a bright red precipitate. Cellular material was removed from the Se(0) particles by using a modified procedure of Laddaga and MacLeod (21). The cell suspensions containing Se(0) were ultrasonicated at 100 W for 2 min and centrifuged at 1,500 ϫ g for 30 min.…”

Section: Methodsmentioning

confidence: 99%

Structural and Spectral Features of Selenium Nanospheres Produced by Se-Respiring Bacteria

Oremland

Herbel

Blum

et al. 2004

Appl Environ Microbiol

409

201

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Certain anaerobic bacteria respire toxic selenium oxyanions and in doing so produce extracellular accumulations of elemental selenium [Se(0)]. We examined three physiologically and phylogenetically diverse species of selenate-and selenite-respiring bacteria, Sulfurospirillum barnesii, Bacillus selenitireducens, and Selenihalanaerobacter shriftii, for the occurrence of this phenomenon. When grown with selenium oxyanions as the electron acceptor, all of these organisms formed extracellular granules consisting of stable, uniform nanospheres (diameter, ϳ300 nm) of Se (0)

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“…The organisms used were: A. haloplanktis 214, variant 3 (ATCC 19855); Vibrio natriegens 107; Photobacterium phosphoreum 404; and Vibrio fischeri MAC401. The sources of these cultures and the methods used to maintain them have been previously described (5,11).…”

Section: Methodsmentioning

confidence: 99%

Relationship between ion requirements for respiration and membrane transport in a marine bacterium

Khanna¹,

DeVoe²,

Brown³

et al. 1984

J Bacteriol

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Intact cells of the marine bacterium Alteromonas haloplanktis 214 oxidized NADH, added to the suspending medium, by a process which was stimulated by Na+ or Li' but not K+. Toluene-treated cells oxidized NADH at three times the rate of untreated cells by a mechanism activated by Na+ but not by Li' or K+. In the latter reaction, K+ spared the requirement for Na+. Intact cells of A. haloplanktis oxidized ethanol by a mechanism stimulated by either Na+ or Li'. The uptake of ot-aminoisobutyric acid by intact cells of A. haloplanktis in the presence of either NADH or ethanol as an oxidizable substrate required Na+, and neither Li' nor K+ could replace it. The results indicate that exogenous and endogenous NADH and ethanol are oxidized by A. haloplanktis by processes distinguishable from one another by their requirements for alkali metal ions and from the ion requirements for membrane transport. Intact cells of Vibrio natriegens and Photobacterium phosphoreum oxidized NADH, added externally, by an Na+-activated process, and intact cells of Vibrio fischeri oxidized NADH, added externally, by a K+-activated process. Toluene treatment caused the cells of all three organisms to oxidize NADH at much faster rates than untreated cells by mechanisms which were activated by Na+ and spared by K+.

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Effects of wash treatments on the ultrastructure and lysozyme penetrability of the outer membrane of various marine and two terrestrial gram-negative bacteria

Cited by 19 publications

References 22 publications

Separation of outer and cytoplasmic membranes of Fibrobacter succinogenes and membrane and glycogen granule locations of glycanases and cellobiase

Separation of outer and cytoplasmic membranes of Fibrobacter succinogenes and membrane and glycogen granule locations of glycanases and cellobiase

Structural and Spectral Features of Selenium Nanospheres Produced by Se-Respiring Bacteria

Relationship between ion requirements for respiration and membrane transport in a marine bacterium

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