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G J Walker

Publications and source records attributed to G J Walker.

87 records · Page 5Linked to original sources
PubMed1976

Metabolism of the reserve polysaccharide of Streptococcus mitior (mitis): is there a second alpha-1,4-glucan phosphorylase?

The alpha-1,4-glucan phosphorylase (alpha-1,4-glucan: orthophosphate glucosyltransferase; EC 2.4.1.1) associated with the particulate cell fraction of Streptococcus mitior strain S3 was compared with the soluble maltodextrin phosphorylase that had been previously isolated from the same organism (Walker et al., 1969). The particulate enzyme was more sensitive to the glycogen content of the cell than the soluble euzyme; its activity was highest when the cells were grown under conditions favoring high glycogen storage. Substrate specificities of the two high activity towards endogenous glycogen, whereas low-molecular-weight maltodextrins were the preferred substrates for the soluble phosphorylase. The purification of the particulate phosphorylase included incubation of the particulate fraction in 160 mM sodium phosphate-10 mM sodium citrate-0.1% (wt/vol) Triton X-100 buffer (pH 6.7) and ion-exchange chromatography on diethylamino-ethyl- Sephadex A-50. The purified enzyme was fully soluble. The value for the purification factor was variable and depended on (i) the substrate used and (ii) whether the synthetic or the degradative reaction was being measured. The solubilization resulted in considerable changes in the properties of the phosphorylase: the pH optimum for activity was raised from 6.0 to 7.0-7.5 and the substrate specificity was altered. Consequently, the purified enzyme bore greater similarity to the soluble maltodextrin phosphorylase. The reported results are best explained in terms of a single phosphorylase, the specificity which is determind by its binding state in the cell. The enzyme acts as a glycogen phosphorylase in the particulate state and as a maltodextrin phosphorylase when soluble. The equilibrium between the two forms is related to the glycogen content of the cells.

Glycogen↗
PubMed1975

The action pattern of Penicillium lilacinum dextranase.

The product distributions resulting from the action of Penicillium lilacinum dextranase on end-labelled oligosaccharides of the isomaltose series have been determined. The initial rates of formation of labelled products were measured for isomaltotriose up to isomalto-octaose, and the molar proportions and radioactivity of the final products from isomaltotriose up to isomaltohexaose were determined. D-Glucose was released only from isomaltotriose and isomaltotetraose, by hydrolysis of the first linkage from the reducing end (linkage 1); the terminal bonds of higher members of the series were not attacked. All oligosaccharides except isomaltotriose were hydrolyzed at more than one linkage. The main points of attack on isomaltotetraose up to isomalto-octaose were at linkage 2, and at the third linkage from the non-reducing end; these two positions coincide for isomaltopentaose. The degradation of isomaltotriose up to isomalto-octaose was entirely hydrolytic. The enzyme also catalyzed an extremely slow, concentration-dependent degradation of isomaltose, and this may have occurred via a condensation to isomaltotetraose, followed by hydrolysis of linkage 1 to give D-glucose and isomaltotriose.

Dextranase↗
PubMed1968

Metabolism of the reserve polysaccharide of Streptococcus mitis. Some properties of a pullulanase.

1. A pullulanase has been separated from cell extracts of Streptococcus mitis. The enzyme was freed from transglucosylase by fractionation with ammonium sulphate. 2. Pullulanase was produced in the absence of inducers, and addition of glucose or maltose to the broth did not increase the yield of enzyme. 3. The pullulanase acted rapidly on alpha-(1-->6)-bonds in substrates having the structure alpha-maltodextrinyl-(1-->6)-maltodextrin, but had no action on isomaltose, 6-alpha-glucosylmaltodextrins or 6-alpha-maltodextrinylglucoses. 4. 6-alpha-Maltotriosylmaltodextrins were hydrolysed over 10 times faster than 6-alpha-maltosylmaltodextrins. 5. The branch linkages of amylopectin phosphorylase limit dextrin, glycogen phosphorylase limit dextrin and glycogen beta-amylase limit dextrin were hydrolysed. The action of pullulanase on amylopectin and glycogen was accompanied by a rise in the iodine stain of 50% and 30% respectively. 6. A reversal of pullulanase action occurred on incubation with high concentrations of maltotriose. Condensation of maltosyl units to form a branched tetrasaccharide occurred less readily. 7. S. mitis pullulanase was rapidly inactivated at temperatures higher than 40 degrees , and the enzyme did not recover activity on storage at room temperature.

Chemical Precipitation↗
PubMed1967

Metabolism of the reserve polysaccharide of Streptococcus mitis. Properties of alpha-(1-->6)-glucosidase, its separation from transglucosylase, and the action of the two enzymes on branched oligosaccharides.

1. An alpha-(1-->6)-glucosidase has been separated from cell extracts of Streptococcus mitis. The enzyme was freed from transglucosylase by adsorption of the latter on retrograded amylose. 2. The enzyme was detected in five of the six strains of S. mitis that were studied; alpha-(1-->6)-glucosidase was not found in strain RB1633, a strain that did not store polysaccharide. 3. The glucosidase could act on compounds in which alpha-glucose is joined through an alpha-(1-->6)-bond to either a maltosaccharide or an isomaltosaccharide. 6(2)-alpha-Glucosylmaltose (panose) and 6(3)-alpha-glucosylmaltotriose were hydrolysed more rapidly and isomaltodextrins more slowly than isomaltose. 4. Transferring activity towards isomaltose and panose was appreciable when the concentration of substrate was 2% or higher. 5. The enzyme had no action on alpha-(1-->4)-glucosidic linkages. 6-alpha-Maltodextrinylglucoses were hydrolysed only after transglucosylase action had attenuated them to isomaltose.

Journal Article↗
PubMed1966

Metabolism of the reserve polysaccharide of Streptococcus mitis: Properties of a transglucosylase.

1. A transglucosylase has been separated from cell extracts of Streptococcus mitis, and has been partially purified by chromatography on DEAE-cellulose. 2. The transglucosylase was present in the six strains of Streptococcus mitis that were examined, and the activity of the enzyme was the same whether the cells had grown on glucose or on maltose. Four of the strains could store intracellular iodophilic polysaccharide when grown on high concentrations of glucose or maltose (1%), but none of the strains stored polysaccharide during growth on 0.1% glucose. The activity of transglucosylase in cell extracts was the same whether or not the cells had stored polysaccharide. 3. The transglucosylase degrades amylose in the presence of a suitable acceptor, transferring one or more glucosyl residues from the non-reducing end of the donor to the non-reducing end of the acceptor. With [(14)C]glucose as acceptor the maltodextrins produced were labelled in the reducing glucose unit only. 4. The enzyme can synthesize higher maltodextrins from maltose and maltotriose. Maltotetraose is disproportionated to give products of sufficient chain length to give a stain with iodine. 5. The action pattern of S. mitis during the degradation of synthetic amylose was shown to be intermediate between the single-chain and multi-chain mechanism.

Journal Article↗
PubMedDate not supplied

No effect of fluid intake on neutrophil responses to prolonged cycling.

Ingesting carbohydrate beverages during prolonged exercise is associated with fewer numbers of circulating neutrophils and attenuated neutrophil functional responses, yet there is little information about the effect of fluid intake alone on immune responses to prolonged exercise. The aim of this study was to examine the influence of regular fluid ingestion compared with no fluid ingestion on plasma cortisol, circulating neutrophil and lipopolysaccharide (LPS)-stimulated neutrophil degranulation responses to prolonged cycling. In a randomized design, nine recreationally active males cycled for 2 h at 65% VO2max on two occasions with either fluid ingestion (lemon-flavoured water, fluid trial) before and during the exercise, or with no fluid intake at all (no fluid trial). Venous blood samples were obtained at rest, immediately after exercise and 1 h after exercise. Immediately after exercise, the plasma cortisol concentration was significantly higher in the no fluid trial than in the fluid trial (592 +/- 62 vs 670 +/- 63 nmol x l(-1), P < 0.05). Circulating numbers of neutrophils increased 4.5-fold (P < 0.01) and LPS-stimulated elastase release per neutrophil decreased 34 +/- 7% (P < 0.01) immediately after exercise; there were no differences between trials. These results suggest that in ambient environmental conditions, fluid ingestion alone has a negligible effect on circulating neutrophil and LPS-stimulated neutrophil degranulation responses to prolonged exercise.

Adult↗