Leukocyte Gcrete Lab Report

Better Essays
To determine optimal pH value for the hydrolysis of p-NPG by leukocyte GCase, we examined effects of variable pH (4.0-5.5) at a constant substrate concentration (5mM). Figure 1 shows the pH activity curves of leukocyte GCase assayed in the presence and absence of 6 mg per ml Sodium taurocholate. In the absence of the pure sodium taurocholate, the activity in the range of pH 4.0-4.8 was considerably lower. However, in the presence of pure Sodium taurocholate, leukocyte GCase was optimally active at pH 5.0, after which the enzyme activity started to decline. Figure 1 also shows that leukocyte glucocerebrosidase activity is completely dependent upon this detergent. Maximum stimulation representing a four- to five-fold increase in activity occurred …show more content…
Grabowski, G.A., and Dagan, A. (1984) Human lysosomal β-glucosidase: Purification by affinity chromatography. Anal. Biochem., 141 (1), 267–279.
37. Maret, A., Salvayre, R., Potier, M., Legler, G., Beauregard, G., and Douste-Blazy, L. (1988) Comparison of Human Membrane-Bound β-Glucosidases: Lysosomal Glucosylceramide-β-Glucosidase and Non-Specific β-Glucosidase, in Lipid Storage Disorders, Springer, pp. 57–61.
38. Fishman, W.H., Springer, B., and Brunetti, R. (1948) Application of an improved glucuronidase assay method to the study of human blood β-glucuronidase. J. Biol. Chem., 173 (2), 449–456.
39. Kato, K., Yoshida, K., Tsukamoto, H., Nobunaga, M., Masuya, T., and Sawada, T. (1960) Synthesisof p-Nitrophenyl β-D-Glucopyranosiduronic Acid and Its Utilization as a Substrate for the Assay of β-Glucuronidase Activity. Chem. Pharm. Bull. (Tokyo), 8 (3), 239–242.
40. Szasz, G. (1967) Comparison between p-nitrophenyl glucuronide and phenolphthalein glucuronide as substrates in the assay of β-glucuronidase. Clin. Chem., 13 (9), 752–759.
41. Harper, B., Luukinen, B., Gervais, J.A., Buhl, K., and Stone, D. (2009) Diazinon General Fact Sheet. Natl. Pestic. Inf. Cent. Or. State Univ. Ext.
…show more content…
Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 72 (1–2), 248–254.
44. Grabowski, G.A., Gatt, S., Kruse, J., and Desnick, R.J. (1984) Human lysosomal β-glucosidase: kinetic characterization of the catalytic, aglycon, and hydrophobic binding sites. Arch. Biochem. Biophys., 231 (1), 144–157.
45. Ašić, A., Bešić, L., Muhović, I., Dogan, S., and Turan, Y. (2015) Purification and characterization of β-glucosidase from Agaricus bisporus (white button mushroom). Protein J., 34 (6), 453–461.
46. Bešić, L., Ašić, A., Muhović, I., Dogan, S., and Turan, Y. (2017) Purification and Characterization of β-Glucosidase from Brassica oleracea. J. Food Process. Preserv., 41 (2).
47. Teugjas, H., and Väljamäe, P. (2013) Selecting β-glucosidases to support cellulases in cellulose saccharification. Biotechnol. Biofuels, 6 (1), 105.
48. Uchiyama, T., Miyazaki, K., and Yaoi, K. (2013) Characterization of a novel β-glucosidase from a compost microbial metagenome with strong transglycosylation activity. J. Biol. Chem., 288 (25),

Related Documents