Studies on the spores of aerobic bacteria. II. The properties of an extracted heat-stable enzyme

Stewart, Babette Taylor; Halvorson, H. O.

doi:10.1016/0003-9861(54)90178-2

Cited by 41 publications

(17 citation statements)

References 10 publications

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“…We have shown i n this study that the exosporium is the probable site of ahnine racemase, adenosine deaminase and hexosamine, which are thought to be important in the germination of spores (Halvorson & Church, 1957). The fact that intact spores rapidly catalyse the racemization of alanine (Stewart & Halvorson, 1958) and the deamination of adenosine (Powell & Hunter, 1956) indicates that the substrates have ready access to these enzymes; this would be afforded if these enzymes are located outside the spore coat. By contrast, the pyrophosphatase (Levinson, Sloan & Hyatt, 1958) has no demonstrable activity in intact spores, and the activity of catalase (Murrell, 1955) is greatly increased by disruption.…”

Section: Discussionmentioning

confidence: 99%

Sonic Disruption of Spores of Bacillus cereus

Berger

Marr

1960

Journal of General Microbiology

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SUMMARY:The sonic disruption of spores of Bacillus cereus gives multi-hit kinetics. The flrst hit destroys the exosporium, which protects the spore body from destruction ; the second hit destroys the spore body. Spores which have been stripped oftheir exosporia are still viable and heat resistant. From the rate of release during genic tmatment it appears that alanine raceITLsse, adenosine deaminase and hexosamine are located in the exosporium while ribosidase is in the spore body. The rate of release of dipicolinic acid does not identify it$ location.Since the discovery of alanine racemase in spores (Stewart & Halvorson, 1953) several other enzymes have been found in preparations of well-cleaned spores (reviewed by Halvorson & Church, 1957). Many of the enzymes in extracts of spores are particulate and heat-stable, but the location of the enzymes within the spore is somewhat obscure. Electron micrographs of thin sections of spores of BaciZZzM cmew (Robinow, 1958) show a large well-defined exosporium as the outermost structure surrounding the spore coat. The electrondense spore coat, in turn, delimits what we shall refer to as the spore body. The particulate enzymes of the spore could be contained within the spore body or could be submicroscopic fragments of the spore coat or of the exosporium. A decision between similar alternatives in Axotobmter virtelartdii has been made by measuring the rates of release of constituents during sonic disruption (Marr & Cota-Robles, 1957). Submicroscopic particles and soluble constituents of the cytoplasm were released at the same rate as the disruption of the cell, while submicroscopic particles derived by disintegration of the envelope were released more slowly. In the application of this technique of Merential release to the biochemical cytology of spores we were somewhat surprised to find that several constituents of the spore are released more Tap'dg than the rate of disruption of the spore body. These constituents appear to be in the exosporium. METHODSOrganism and w h r a l metho&. B Spores were harvested in a Sharpies centrifuge and washed 6-10 times with distilled water or phosphate buffer until microscopic examination indicated the absence of debris from vegetative cells. The spores were suspended in 0.05 M-phosphate buffer (pH 6.8) to a concentration of 8 x loo spores/ml. with less than 5 % gemhated spores. Sm& disruptim. Of the above spore suspension 50ml. were treated at 75 scousticd watb in a Raytheon 10 kc. sonic oscillator at 0 ' 4 ' and in a gas atmosphere of H , . At various times, 8 or 5 ml. samples were removed and were replaced by an equal volume of buffer. Corrections were made for the progressive dilution resulting from this sampling. Release of constituents was determined by centrifuging portions of the samples at 10,WO.g for 15 min., which was sufficient to sediment residual spores and fragments of microscopic dimensions. Release was judged by failure to sediment in this centrifugation. All assays except that for deaminase were made on the residue.Tu...

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Section: Discussionmentioning

confidence: 99%

Sonic Disruption of Spores of Bacillus cereus

Berger

Marr

1960

Journal of General Microbiology

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“…Spores were formed in G medium as described by Stewart and Halvorson (1953), and were washed in triplicate by centrifugation at 7,000 × g at 4℃ for 10 min in 0.85% sodium chloride (Nacalai Tesque, Inc., Kyoto, Japan) solution. The spore suspension was heated at 80℃ for 30 min in order to inactivate vegetative cells.…”

Section: Methodsmentioning

confidence: 99%

Effects of Carbonation with Heating on Germination of Bacillus subtilis Spores

Noma

Yamashita

Klangpetch

et al. 2011

FSTR

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The effects of carbonation with heating (CH) on germination of Bacillus subtilis spores were investigated. Treatment conditions for CH and heat treatment alone were set to obtain an approximate 1 log reduction in viable count. Pre-treatment of spores with CH at 80℃ and 5 MPa for 30 min significantly decreased their heat resistance to a subsequent heating process at 90℃ for 30 min, as compared with pretreatment by heat alone at 90℃ for 30 min. Treatment with CH also decreased refractility and enhanced DAPI staining when compared with heat treatment alone, thus suggesting that CH effectively initiates and stimulates germination of B. subtilis spores.

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“…First, the organisms may contain a histidine racemase comparable with the alanine racemase of Streptococcus faecalis and other bacteria (Wood & Gunsalus, 1951;Stewart & Halvorson, 1953;Marr & Wilson, 1954), the methionine racemase of a pseudomonad (Kallio & Larson, 1955) or the glutamate racemase of Lactobacillus arabinosus (Narrod & Wood, 1952). Secondly, D-histidine may be attacked by a D-amino acid oxidase, yielding P-imidazole pyruvate which, by transamination, could yield L-histidine.…”

Section: Discussionmentioning

confidence: 99%

The Adaptive Degradation of L-Histidine by Paracolobactrum aerogenoides

Tristram¹

1960

Journal of General Microbiology

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SUMMARYThe cliaracteristics of an induced enzyme system responsible for the degradation of L-histidine by a soil organism were studied. Induction could be achieved by exposing cells to L-histidine, D-histidine, urocanate, p-alanyl-L-histidine or L-histidyl-L-histidine, but not by exposure to a variety of other imidazoles. The kinetics of induction by L-and D-histidine differed considerably and the D-isomer was shown to be metabolized only very slowly by fully induced cells. Chloramphenicol and DL-p-fluorophenylalanine strongly inhibited enzyme synthesis, whilst a variety of purine and pyrimidine analogues were without effect. Nitrogen starvation of noninduced cells decreased but did not completely prevent the appearance of the enzyme system on induction with D-histidine. Non-induced cells possess a low level of ' basal ' enzymes before exposure to an inducing substance.

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Studies on the spores of aerobic bacteria. II. The properties of an extracted heat-stable enzyme

Cited by 41 publications

References 10 publications

Sonic Disruption of Spores of Bacillus cereus

Sonic Disruption of Spores of Bacillus cereus

Effects of Carbonation with Heating on Germination of Bacillus subtilis Spores

The Adaptive Degradation of L-Histidine by Paracolobactrum aerogenoides

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