Journal of comparative physiology

, Volume 115, Issue 2, pp 241–253 | Cite as

Trehalase transformation in silkworm midgut during metamorphosis

  • Motoyuki Sumida
  • Okitsugu Yamashita
Article

Summary

The particulate trehalase from silkworm larval midgut was effectively solubilized by repeated freezing and thawing, and by incubation with snake venom and non-ionic detergents (Lubrol PX and WX and Triton X-100). With solubilization the activity was enhanced and the activation behaviour was dependent upon the developmental stage of silkworms, being highest (up to about 3-fold) at the spinning stage.

When chromatographed on DEAE-cellulose columns separately, the enzyme solubilized by freezing and thawing and the soluble trehalases from feeding larval midgut were respectively eluted as single peaks, P I and P II. However, both P I and P II trehalases were demonstrated after solubilization of the particulate fraction from feeding larvae with Triton X-100, or after treatment of the midgut of spinning larvae by freezing and thawing.

The apparent molecular weights of P I and P II trehalases as estimated by Sephadex G-200 chromatography were about 70,000 and 140,000, respectively. The optimum pH was 6.0 for P I and about 5.0 for P II trehalase. TheKm values were about 1.0 mM for P I trehalase and 0.30 mM for P II trehalase.

These results suggest that in feeding larval midgut there are two types of trehalase which are distinguishable from each other by intracellular localization, protein nature and kinetic properties. Furthermore, when the midgut undergoes metamorphosis, the P I enzyme found predominantly in feeding stages seems to be transformed to the P II enzyme via an intermediate form (Ppt-P II) with transitional properties.

Keywords

Enzyme Activation Behaviour Single Peak Protein Nature Particulate Fraction 

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Copyright information

© Springer-Verlag 1977

Authors and Affiliations

  • Motoyuki Sumida
    • 1
  • Okitsugu Yamashita
    • 1
  1. 1.Laboratory of Sericultural Science, Faculty of AgricultureNagoya UniversityChikusa, NagoyaJapan

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