Characteristics of alkaline and acid phosphatase in <i>Spirometra erinacei</i>

, Kwak, K H; , Kim, C H

doi:1996.34.1.69

Korean J Parasito > Volume 34(1):1996 > Article

Original Article


Korean J Parasitol. 1996 Mar;34(1):69-77. English. Published online Mar 20, 1996. http://dx.doi.org/10.3347/kjp.1996.34.1.69
Copyright © 1996 by The Korean Society for Parasitology


Characteristics of alkaline and acid phosphatase in Spirometra erinacei
Kee-Hoon Kwak and Chang-Hwan Kim^*
Department of Biology, College of Natural Sciences Gyeongsang National University, Chinju 660-701, Korea.

Received November 27, 1995; Accepted January 16, 1996.
Abstract
This study was done to investigate the enzyme-histochemical localization and characteristics of alkaline and acid phosphatase related with metabolism in sparganum and adult of Spirometra erinacei. By the enzyme-histochemical assay, the alkaline and acid phosphatases were localized in the tegument and subtegumental musculature of sparganum and adult, but not in the parenchyma. The activities of alkaline phosphatase were stronger in the tegument than in the subtegumental musculature, and activities of acid phosphatase were stronger in the tegument of adults than those of sparganum. The 2 isozymes of alkaline and acid phosphatases were separated from s- sparganum (from snake) and r-sparganum (from experimentally infected rats) respectively, but 4 isozymes of Alp and 3 isozymes of Acp were separated from adult worms by electrophoresis. In isozyme Alp, the 66 kDa was the common isozyme, but 130 kDa isozyme of Acp was the common isozyme in spargana and adult worms. By isoelectrofocusing, 4 isozymes (PI 7.9, 7.7, 6.5 and 6.3) and 2 isozymes (PI 7.9 and 7.7) of alkaline phosphatase were separated from adults and spargana, respectively. In the stability against heat, activity of alkaline phosphatase was denatured perfectly after heating at 90℃ for 40 seconds. The optimum pH and temperature for activity of alkaline phosphatase were about pH 10 and 50℃, respectively. The maximum activity (unit) of alkaline phosphatase was 22.0 in s-sparganum, 25.0 in r-sparganum and 215.0 in adult worms, so that the maximum activity was revealed higher in adults than spargana. As the result from above, we observed that alkaline and acid phosphatases were functioned mainly in the tegument and subtegumental musculature, and the isozymes of phosphatase were activated differently according to habitat of the parasites. The spargana and adult worms carry out the parasitism by adapting themselves to parasitic circumstance with these enzymes.

Figures

Fig. 1
Histochemical localization of alkaline phosphatase on the sparganum and adult worm of S. erinacei. a. Snake sparganum (80 ×) b. Snake sparganum (400 ×) c. Rat sparganum (80 ×) d. Rat sparganum (400 ×) e. Immature proglottid (80 ×) f. Immature proglottid (400 ×) g. Mature proglottid (80 ×) h. Mature proglottid (400 ×) i. Gravid proglottid (80 ×) j. Gravid proglottid (400 ×) T, tegument; SM, subtegumental musculature; PM, parenchymal musculature; CC, calcareous corpuscles; TS, testis; V, vitelline gland; EG, egg; N, nerve trunk.

Fig. 2
Histochemical localization of acid phosphatase on the sparganum and adult worm of S. erinacei. a. Snake sparganum (80 ×) b. Snake sparganum (400 ×) c. Rat sparganum (80 ×) d. Rat sparganum (400 ×) e. Immature proglottid (80 ×) f. Immature proglottid (400 ×) g. Mature proglottid (80 ×) h. Mature proglottid (400 ×) i. Gravid proglottid (80 ×) j. Gravid proglottid (400 ×) T, tegument; SM, subtegumental musculature; PM, parenchymal musculature; CC, calcareous corpuscles; EG, egg.


	Fig. 3 Isozyme patterns of alkaline phosphatase on the sparganum and adult worm of S. erinacei. SS, snake sparganum; RS, rat sparganum; AD, adult; TD, tracking dye; kDa, kilo-dalton; M, marker protein.


	Fig. 4 Isozyme patterns of acid phosphatase on the sparganum and adult worm of S. erinacei. SS, snake sparganum; RS, rat sparganum; AD, adult; TD, tracking dye; kDa, kilo-dalton; M, marker protein.


	Fig. 5 Isozyme patterns of alkaline phosphatase by IEF in adult worm. SP, rat sparganum; AD, adult; M, marker protein; PI, isoelectric point.


	Fig. 6 The stability against heat treatment on the isozyme of alkaline phosphatase in adult worm, heating at 90℃ for 10, 20, 30, 40 second, respectively. 0, control; 10, 10 sec.; 20, 20 sec.; 30, 30 sec.; 40, 40 sec.


	Fig. 7 The effect of pH on the activity of alkaline phosphatase in r-sparganum and adult worm. ●—●, adult; ○—○, sparganum.


	Fig. 8 The effect of temperature on the activity of alkaline phosphatase in r-sparganum and adult worm. ●—●, adult; ○—○, sparganum.

Tables


	Table 1 Relative amount of alkaline phosphatase activity on the sparganum and adult worm of S. erinacei


	Table 2 Relative amount of acid phosphatase activity on the sparganum and adult worm of S. erinacei


	Table 3 Enzyme activities of alkaline phosphatase on the sparganum and adult worm of S. erinacei (mean ± SE)

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