Abstract
beta-Galactosidase (EC 3.2.1.23) from persimmon fruit was purified 114-fold with a 15% yield using Sephadex G-100 gel filtration, CM-Sephadex ion exchange, and Sephacryl S-200 gel filtration chromatography, with subsequent electroelution from nondenaturing polyacrylamide gel electrophoresis (PAGE) gels. The estimated molecular mass of the native beta-galactosidase by Sephacryl S-200 was 118 kD. After sodium dodecyl sulfate-PAGE of the enzyme electroeluted from native gels, two subunits with estimated molecular masses of 34 and 44 kD were observed, suggesting that the native enzyme was an aggregate of several subunits. Amino acid composition and N-terminal amino acid sequences of the two major subunits were different.
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- Bhown A. S., Bennett J. C. High-sensitivity sequence analysis of proteins recovered from sodium dodecyl sulfate gels. Methods Enzymol. 1983;91:450–455. doi: 10.1016/s0076-6879(83)91042-x. [DOI] [PubMed] [Google Scholar]
- Giovannoni J. J., DellaPenna D., Bennett A. B., Fischer R. L. Expression of a chimeric polygalacturonase gene in transgenic rin (ripening inhibitor) tomato fruit results in polyuronide degradation but not fruit softening. Plant Cell. 1989 Jan;1(1):53–63. doi: 10.1105/tpc.1.1.53. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kim J., Gross K. C., Solomos T. Characterization of the Stimulation of Ethylene Production by Galactose in Tomato (Lycopersicon esculentum Mill.) Fruit. Plant Physiol. 1987 Nov;85(3):804–807. doi: 10.1104/pp.85.3.804. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Priem B., Gitti R., Bush C. A., Gross K. C. Structure of ten free N-glycans in ripening tomato fruit. Arabinose is a constituent of a plant N-glycan. Plant Physiol. 1993 Jun;102(2):445–458. doi: 10.1104/pp.102.2.445. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Priem B., Gross K. C. Mannosyl- and Xylosyl-Containing Glycans Promote Tomato (Lycopersicon esculentum Mill.) Fruit Ripening. Plant Physiol. 1992 Jan;98(1):399–401. doi: 10.1104/pp.98.1.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Raghothama K. G., Lawton K. A., Goldsbrough P. B., Woodson W. R. Characterization of an ethylene-regulated flower senescence-related gene from carnation. Plant Mol Biol. 1991 Jul;17(1):61–71. doi: 10.1007/BF00036806. [DOI] [PubMed] [Google Scholar]
- Ranwala A. P., Suematsu C., Masuda H. The Role of beta-Galactosidases in the Modification of Cell Wall Components during Muskmelon Fruit Ripening. Plant Physiol. 1992 Nov;100(3):1318–1325. doi: 10.1104/pp.100.3.1318. [DOI] [PMC free article] [PubMed] [Google Scholar]