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Journal ArticleDOI

Multiple forms of β-galactosidase from the germinating seeds of Vigna radiata

TL;DR: Four forms of β-galactosidase were isolated and partially purified from the cotyledons of germinating seeds of Vigna radiata by ammonium sulphate fractionation and ion exchange chromatography through DEAE-cellulose and CM- cellulose columns, showing similar pH optima and temperature optima but differed from each other in ionic charge, kinetic parameters, activation energy and sensitivity towards heat.
About: This article is published in Phytochemistry.The article was published on 1990-01-01. It has received 11 citations till now. The article focuses on the topics: Galactoside.
Citations
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Journal ArticleDOI
01 Apr 1993-Planta
TL;DR: Results indicate that the β-galactosidase of this study is solely responsible for the removal of galactose from the cell wall during ripening, and its in-vivo activity must be much greater than that observed in- vitro.
Abstract: A β-galactosidase (EC 3.2.1.23) capable of degrading a number of fruit cell-wall polysaccharides in vitro, was isolated from ripening kiwifruit (Actinidia deliciosa [A. Chev.] C.F. Liang et A.R. Ferguson cv. Hayward). The enzyme has a molecular weight of approximately 60 kDa by gel permeation and consists of several basic isoforms. Several polypeptides were enriched during purification, with 33-, 46- and 67-kDa bands being predominant after sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The optimum activity of the enzyme against p-nitrophenyl-β-d-galactopyranoside was at pH 3.2, but against a galactan purified from kiwifruit cell walls, it was at pH 4.9. The enzyme was specific for galactosyl residues in the β-configuration, releasing galactose from a variety of kiwifruit cell-wall polysaccharide fractions including cell wall material, Na2CO3-soluble pectin, high-molecular-weight galactan, xyloglucan, and galactoglucomannan. A galactosylated glucuronomannan found throughout the kiwifruit plant was also a substrate for the enzyme. The results indicate that the enzyme attacks the non-reducing end of galactose side chains, cleaving single galactose residues which may be attached to the 2, 3, 4, or 6 position of the aglycone. Activity of the enzyme in-vitro was too low to account for the total loss of galactose from the cell walls during ripening. If the β-galactosidase of this study is solely responsible for the removal of galactose from the cell wall during ripening then its in-vivo activity must be much greater than that observed in-vitro.

128 citations


Cites background from "Multiple forms of β-galactosidase f..."

  • ...Others have also reported difficulty in maintaining active, purified 13-galactosidase in storage (Burns 1990; Dick et al. 1990; Kundu et al. 1990), and have observed the continued appearance of low-molecular-weight bands in SDS-PAGE of purified 13-galactosidase (Edwards et al....

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  • ...Other authors have found differences in pH optima for various isoforms of 13-galactosidase against synthetic substrates (Pressey 1983; Burns 1990; Dick et al. 1990; Kundu et al. 1990)....

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Journal ArticleDOI
01 Jan 1994-Planta
TL;DR: The exo-galactanase is clearly a key enzyme in galactan mobilisation and may be the sole activity involved in depolymerising the dominant (1 → 4)-β-Galactan component of the cell wall.
Abstract: The main polysaccharide component of the thickened cell walls in the storage parenchyma of Lupinus angustifolius L. cotyledons is a linear (1 → 4)-β-linked d-galactan, which is mobilised after germination (L.A. Crawshaw and J.S.G Reid, 1984, Planta 160, 449–454). The isolation from the germinated cotyledons of a β-d-galactosidase or exo-(1 → 4)-β-d-galactanase with a high specificity for the lupin galactan is described. The enzyme, purified using diethylaminoethyl-cellulose, carboxymethyl-cellulose and affinity chromatography on lactose-agarose, gave two bands (major 60 kDa, minor 45 kDa) on sodium dodecyl sulphate-gel electrophoresis, and two similar bands on isoelectric focusing (major, pI 7.0, minor pI 6.7, both apparently possessing enzyme activity). The minor component cross-reacted with an antiserum raised against, and affinity-purified on, the major band. Both components had a common N-terminal sequence. The minor component was probably a degradation product of the major one. The enzyme had limited β-galactosidase action, catalysing the hydrolysis of p-nitrophenyl-β-d-galactopyranoside and (1→ 4)- and (1 → 6)-β-linked galactobioses. Lactose [β-d-galactopyranosyl-(1 → 4)-d-glucose] was hydrolysed only very slowly and methyl-β-d-galactopyranoside not at all. Lupin galactan was hydrolysed rapidly and extensively to galactose, whereas other cell-wall polysaccharides (xyloglucan and arabinogalactan) with terminal non-reducing β-d-galactopyranosyl residues were not substrates. A linear (1 → 4)-β-linked galactopentaose was hydrolysed efficiently to the tetraose plus galactose, but further sequential removals of galactose to give the tetraose and lower homologues occurred more slowly. Galactose, γ-galactonolactone and Cu+2 were inhibitory. No endo-β-d-galactanase activity was detected in lupin cotyledonary extracts, whereas exo-galactanase activity varied pari passu with galactan mobilisation. Exo-galactanase protein was detected, by Western immunoblotting of cotyledon extracts, just before the activity could be assayed and then increased and decreased in step with the enzyme activity. The exo-galactanase is clearly a key enzyme in galactan mobilisation and may be the sole activity involved in depolymerising the dominant (1 → 4)-β-galactan component of the cell wall.

83 citations

Journal ArticleDOI
TL;DR: Double immunodiffusion analysis indicated thatbeta-galactosidases I, II, III and V are immunologically similar to each other, while beta-galactsosidase IV shares partially identical antigenic determinants with the other four isoforms.

70 citations

Journal ArticleDOI
TL;DR: The β-galactosidase isolated from cotyledons of germinating seeds of Copaifera langsdorffii seems to perform a key role in xyloglucan degradation since it is responsible for the retrieval of a major sterical hindrance for further hydrolysis of the oligosaccharides and therefore essential for completion of xylglucan mobilisation.

43 citations

Journal ArticleDOI
TL;DR: β -Galactosidase activity in coffee berries showed a progressive increase of more than four-fold as the fruit developed from the immature to ripe stage, with a slight decrease in fully ripe fruit, suggesting that β-galactsidase plays a role in cell wall degradation such as occurs during fruit ripening.

37 citations

References
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Journal Article
TL;DR: Procedures are described for measuring protein in solution or after precipitation with acids or other agents, and for the determination of as little as 0.2 gamma of protein.

289,852 citations

Journal ArticleDOI
15 Aug 1970-Nature
TL;DR: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products.
Abstract: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products. Four major components of the head are cleaved during the process of assembly, apparently after the precursor proteins have assembled into some large intermediate structure.

232,912 citations

Journal Article
01 Jan 1970-Nature
TL;DR: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products as mentioned in this paper.
Abstract: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products. Four major components of the head are cleaved during the process of assembly, apparently after the precursor proteins have assembled into some large intermediate structure.

203,017 citations

Journal ArticleDOI
TL;DR: The results are similar to those of previous studies, where the objective was to establish a cause-and-effect relationship, rather than a straightforward relationship between the number of cells and the content of the molecule.
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3,756 citations