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Cytotechnology

, Volume 33, Issue 1–3, pp 47–52 | Cite as

Purification and characterization of α-D-galactosidase produced by ADG cell line established from abalon digestive gland

  • Ken-ichi Kusumoto
  • Sanetaka ShirahataEmail author
  • Yuto Kamei
Article

Abstract

ADG cell line was established from an abalonedigestive gland and previously characterized. ADGcells have the potential to grow in protein-freeculture and secrete l3 types of glycosidases. Inthis article, we determined the origin of ADG cell line,using electron microscopy, and purified a glycosidasesecreted by these cells. The electron microscopicanalysis showed that ADG cell line contains severalnuclei, which suggests that they may be derived fromprotist cells. Moreover, α-D-galactosidasethat hydrolyzes p-nitorophenyl galactopyranosidewas purified 130-fold from the spent culture medium ofADG cells. The molecular weight of the enzyme,determined by sodium dodecyl sulfate polyacrylamidegel electrophoresis and gel filtration analysis, wasshown to be 43 and 42 kDa, respectively, and itappeared to consist of a single polypeptide chain. The purified enzyme preparation was practically freefrom other glycosidases secreted from the cells. Catalytic activity was optimal at pH 5.5 and at atemperature of 37 °C. The enzyme was also the most stable at pH 5.5.

abalone digestive gland cell protein-free culture α-D-galactosidase 

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Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • Ken-ichi Kusumoto
    • 1
  • Sanetaka Shirahata
    • 2
    Email author
  • Yuto Kamei
    • 3
  1. 1.Cellular Regulation Technology Laboratory, Graduate School of Genetic Resources TechnologyKyushu UniversityFukuokaJapan
  2. 2.Cellular Regulation Technology Laboratory, Graduate School of Genetic Resources TechnologyKyushu UniversityFukuokaJapan
  3. 3.Marine and Highland Bioscience CenterSaga UniversitySagaJapan

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