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  • Title: Measurement of caprine plasma beta-mannosidase with a p-nitrophenyl substrate.
    Author: Cavanagh K, Dunstan RW, Jones MZ.
    Journal: Am J Vet Res; 1983 Apr; 44(4):681-4. PubMed ID: 6869965.
    Abstract:
    Experimental conditions for measuring caprine plasma beta-D-mannosidase activity are described with p-nitrophenyl-beta-D-mannopyranoside as a substrate. The plasma enzyme was stable for 3 months at -20 C or 1 week at 4 C. The optimal pH for activity was 5.0 in citrate-phosphate or acetate buffer. Enzyme activity was linear with time up to 24 hours at 37 C, but incubation of plasma at 56 C for 5 minutes resulted in loss of all activity. The apparent Michaelis-Menten constant (Km) for the plasma enzyme was 10.0 mM. Plasma beta-mannosidase from clinically normal and beta-mannosidosis carrier goats did not differ with respect to pH optimum, heat stability, or Km. The coefficient of variation for the assay, determined by assaying a plasma pool over a 3-month period, was 10.7% (mean: 115 nmole of p-nitrophenol formed/hour/ml of plasma). The assay described can be used to evaluate plasma beta-mannosidase measurements as a test for detecting carriers of caprine beta-mannosidosis, a newly described lysosomal storage disease.
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