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JXB Advance Access originally published online on July 28, 2003
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Journal of Experimental Botany, Vol. 54, No. 390, pp. 2045-2052, September 1, 2003
© 2003 Oxford University Press

Multiple forms of endo-1,4-ß-glucanases in the endosperm of Euphorbia heterophylla L.

Received 20 March 2003; Accepted 28 May 2003

Cecilia N. K. Suda1 and Jarbas F. Giorgini*,2

1 Departamento de Bioquímica e Imunologia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, 14049-900, Brazil
2 Departamento de Biologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, 14040-901, Brazil

* To whom correspondence should be addressed. Fax: +55 16 602 3666. E-mail: jarbas{at}ffclrp.usp.br

Germinating seeds of Euphorbia heterophylla L. contain endo-1,4-ß-glucanases which degrade carboxymethylcellulose (CMC). The activity decreased approximately 66% in extracts of endosperm containing isopropanol or ethanol. The endoglucanases were isolated from endosperm extracts using ammonium sulphate fractionation followed by Sephacryl S-100-HR chromatography resulting in two main peaks: I and II. Peak I endoglucanase was further purified about 15-fold on DEAE-Sephadex A50 and then by affinity chromatography (CF11-cellulose). Peak II endoglucanases were further purified 10-fold on CM-cellulose chromatography. The results indicated the occurrence of a 66 kDa endoglucanase (fractionated by SDS-PAGE and visualized by activity staining using Congo Red). Several acidic (pI 3.0 to 5.7) and basic (pI 8.5 to 10.0) forms from both peaks which differed in their capacities for degrading CMC or xyloglucans from Copaifera langsdorffii or Hymenaea courbaril were detected.

Key words: Endoglucanase, Euphorbia heterophylla, Euphorbiaceae, seed germination.


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