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

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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.

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References

  • Awaji M, Machii A, Nakamura HK and Wada KT (1989) Characterization of a zoosporic fungus appearing in primary cultures of the kuruma prawn midgut gland. Bull Natl Res Inst Aquaculture 15: 19–27.

    Google Scholar 

  • Barr DJ (1981) The phylogenetic and taxonomic implications of flagellar rootlet morphology among zoosporic fungi. Biosystems 14: 359–370.

    Google Scholar 

  • Barr DJ (1988) How modern systematics relates to the rumen fungi. Biosystems 21: 351–356.

    Google Scholar 

  • Chien SF and Lin-Chu M (1991) The conversion of group B red blood cells into group O by an α-D-galactosidase from taro (Colocasia esculenta). Carbohydr Res 217: 191–200.

    Google Scholar 

  • Civas A, Eberhard R, Le Dizet P and Petek F (1984) Glycosidases induced in Aspergillus tamarii. secreted α-D-galactosidase and β-D-mannanase. Biochem J 219: 857–863.

    Google Scholar 

  • Dhar M, Mitra M, Hata J, Butnariu O and Smith D (1994) Purification and characterization of Phaseolus vulgaris α-Dgalactosidase isozymes. Biochem Mol Biol Int 34: 1055–1062.

    Google Scholar 

  • Haibach F, Hata J, Mitra M, Dhar M, Harmata M, Sun P and Smith D (1991) Purification and characterization of a Coffea canephora α-D-galactosidase isozymes. Biochem Biophys Res Commun 181: 1564–1571.

    Google Scholar 

  • Ishikawa Y and Saga N (1990) Study on the breeding of marine algae applying tissue culture technique. Soshiki Baiyou 16: 173–177.

    Google Scholar 

  • Kawashima Y and Tokuda H (1990) Protoplast isolation from several macroalgae and cell fusion. Soshiki Baiyou 16: 157–161.

    Google Scholar 

  • Kuras R and Wollman FA (1994) The assembly of cytochrome b6/f complexes: an approach using genetic transformation of the green alga Chlanydomonas reinhardii. EMBO J 13: 1019–1027.

    Google Scholar 

  • Kusumoto K, Shirahata S, Katakura Y, Murakami H and Kamei Y (1997) Establishment of an abalone digestive gland cell line secreting various glycosidases in protein-free culture. Cytotechnology 24: 169–175.

    Google Scholar 

  • Nakanishi K and Saga N (1990) Production of useful compounds in marine algae cell culture. Soshiki Baiyou 16: 177–180.

    Google Scholar 

  • Rochaix J and Dillewijin IV (1982) Transformation of the green alga Chlanydomonas reinhardii with yeast DNA. Nature 296: 70–72.

    Google Scholar 

Download references

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Correspondence to Sanetaka Shirahata.

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Kusumoto, Ki., Shirahata, S. & Kamei, Y. Purification and characterization of α-D-galactosidase produced by ADG cell line established from abalon digestive gland. Cytotechnology 33, 47–52 (2000). https://doi.org/10.1023/A:1008175226819

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  • DOI: https://doi.org/10.1023/A:1008175226819

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