Adenosine deaminase and ribosidase in spores of <i>Bacillus cereus</i>

Powell, Joan F.; Hunter, J. R.

doi:10.1042/bj0620381

Cited by 56 publications

(27 citation statements)

References 14 publications

(5 reference statements)

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“…After conversion to a form which does not sediment in high-speed centrifugation, both enzymes become more sensitive to hating, particularly the deaminase, in agreement with the findings of stewart & Halvorson (1954) and Powell & Hunter (1956). It seems unlikely that the heat resistance of those external enzymes is related to the heat J .…”

Section: Discussionsupporting

confidence: 80%

“…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%

“…G. Maw resistance of the spore proper but rather is conferred by the particulate complex which contains the enzyme. The heat resistance of ribidase, which was found in the spore coat, also appears to be independent of the integrity of the spore (Powell & Hunter, 1956). The pyrophosphatase and catalase, which appear to be inside the spore body, are not destroyed by heating the intact spore but are destroyed by heating crushed spores (Levinson, Sloan & Hyatt, 1958; MurreU, 1955).…”

Section: Discussionmentioning

confidence: 99%

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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: Discussionsupporting

confidence: 80%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Sonic Disruption of Spores of Bacillus cereus

Berger

Marr

1960

Journal of General Microbiology

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“…In this instance, however, inosine could be replaced by guanosine, but not by adenosine nor xanthosine. It has been suggested [16] that the germination ability of adenosine is due to its conversion to inosine by specific enzyme, adenosine deaminase. This possibly means that spores of the organism is devoid of this enzyme, but this explanation does not account for the germination ability of guanosine.…”

Section: Discussionmentioning

confidence: 99%

The Germination Requirements of Spores of Clostridium botulinum Type E

Ando

1971

Japanese Journal of Microbiology

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The requirements for, and conditions influencing, the germination of spores of Clostridium botulinum type E were studied in chemically defined media. Spores of this organism germinated rapidly and completely in various combinations of L-alanine and one of such carbon sources as sugars, lactate, and ribosides in the presence of bicarbonate. The most effective compounds tested were glucose, L-and D-lactate, and inosine. The formation of lactic acid was found to occur during germination of spores in a combination of L-alanine and glucose. Furthermore, the germination in this medium was strongly inhibited by fluoride or iodoacetate. The germination of spores was also initiated with high concentrations (100 mm or more) of L-alanine or certain other amino acids alone in the presence of bicarbonate at pH 9.0. The spores germinating in L-alanine alone and those in L-alanine plus other compounds were compared as to the minimal concentration of L-alanine required, pH optimum, temperature optimum for heat-activation, bicarbonate requirement, and effects of various inhibitors. Significant differences among these characteristics led to a conclusion that different metabolic pathways, operating selectively each germinative compound used, might be involved in the germination of spores of the organism. The mode of action of combination in stimulating germination by the different germinative compounds has been discussed.

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“…In B. cereus spores and vegetative cells anenzyme activity catalyzing the deamination of adenosine was originally found by Powell and Hunter [7] and by Powell and Strange [8].…”

Section: Introductionmentioning

confidence: 99%