NIGERIAN JOURNAL OF SCIENCE AND ENVIRONMENT
Journal of the Faculties of Science and Agriculture, Delta State University, Abraka, Nigeria

ISSN: 1119-9008
DOI: 10.5987/UJ-NJSE
Email: njse@universityjournals.org


PARTIAL PURIFICATION AND CHARACTERIZATION OF β-DGALACTOSIDASE FROM PLANTAIN FRUIT (Musa paradisiaca L.)

DOI: 10.5987/UJ-NJSE.16.039.1   |   Article Number: E69F3C5   |   Vol.11 (1) - September 2012

Authors:  Agoreyo B.O and Golden K.D

Keywords: Musa paradisiaca, plantain, galactosidase/β-D-Galactosidase in plantain

β-D-Galactosidase (EC 3.2.1.23) from ripe plantain fruit was partially purified and characterized. Purification was carried out using ammonium sulphate precipitation and gel-filtration on Sephadex G25-150 and G-25M. The enzyme displayed activity against p-nitrophenyl-β-D-galactopyranoside, with a Km of 1 mM. It was inhibited by galactose and mercuric chloride. Galactose was a noncompetitive inhibitor, while mercuric chloride was an uncompetitive inhibitor.

Ali, Z.M., Armugam, S. and Lazan, H.

(1995). β-Galactosidase and its significance

in ripening mango fruit. Phytochemistry

38:1109-1114.

Ali, Z.M., Ng, S.Y., Othman, R., Goh, L.Y.

and Lazan, H. (1998). Isolation, characterization

and significance of papaya β-

galactanases to cell wall modification

and fruit softening during ripening. Physiologia

Plantarum 104:105-115.

Balasubramaniam, S., Lee, H.C., Lazan,

H., Othman, R. and Ali, Z.M. (2005).

Purification and properties of a β-

galactosidase from carambola fruit with

significant activity towards cell wall polysaccharides.

Phytochemistry 66:153-163.

Biles, C.L., Bruton, B.D., Russo, V. and

Wall, M.M. (1997). Characterization of β-

galactosidase isoenzymes of ripening peppers.

Journal of the Science of Food and

Agriculture 75:237-243.

Brenchley, J. (1996). Psychrophilic microorganisms

and their cold – active enzymes.

Journal of Industrial Microbiology 17:

432 - 437

Brummell, D.A. and Harpster, M.H.

(2001). Cell wall metabolism in fruit softening

and quality and its manipulation in

transgenic plants. Plant Molecular Biology

47:311-340.

Carey, A.T., Holt, K., Picard, S., Wilde, R.,

Tucker, G.A., Bird, C.R., Schuch,

W. and Seymour, G.B. (1995). Tomato

exo-(1-4)-β-D- galactanase: Isolation,

changes during ripening in normal and

mutant tomato fruit, and characterization

of a related cDNA clone. Plant Physiology

108:1099 - 1107.

Carpita, N.C. and Gibeaut, D.M. (1993).

Structural models of primary cell walls in

flowering plants: consistency of molecular

structure with the

physical properties of the walls during

growth. The Plant Journal 3: 1-30.

Carrington, C.M.S. and Pressey, R. (1996).

β-Galactosidase II activity in relation to

changes in cell wall galactosyl composition

during tomato ripening. Journal of

American Society of Horticultural Science

121:132-136.

Chun, J-P. and Huber, D.J. (2000). Reduced

levels of β-subunit protein influence tomato

fruit firmness, cell-wall ultrastructure,

and PG2-mediated pectin hydrolysis

in excised pericarp tissue. Journal of Plant

Physiology 157:153-160.

Cooper, W., Bouzayen, M., Hamilton, A.,

Barry, C., Rossall, S. and Grierson, D.

(1998). Use of transgenic plants to study

the role of ethylene and polygalacturonase

during infection of tomato fruit by Colletotrichum

gloeosporioides. Plant Pathology

47:308-316.

Cosgrove, D.J. (2001). Wall structure and

wall loosening. A look backwards and forwards.

Plant Physiology 125:131-134.

Crookes, P.R. and Grierson, D. (1983). Ultrastructure

of tomato fruit ripening and

the role of polygalacturonase isozymes in

cell wall degradation. Plant Physiology

72:1088-1093.

deVeau, E.J.I., Gross, K.C., Huber, D.J.

and Watada, A.E. (1993). Degradation

and solubilization of pectin by β-

galactosidase purified from avocado meso-

carp. Physiologia Plantarum 87:279-288.

Dick, A.J., Opoku-Gyamfua, A. and

DeMarco, A.C.(1990). Glycosidases of

apple fruit: a multifunctional β-

galactosidase. Physiologia Plantarum

80: 250-256.

Golden, K.D., John, M.A. and Kean, E.C.

(1993). β-Galactosidase from Coffea arabica

and its role in fruit ripening. Phytochemistry

34: 355- 360.

Grierson, D. and Schuch, W. (1993). Control

of ripening. Philosophical Transactions

of the Royal Society of London, Series

B 342: 241-250.

Gross, K.C. (1984). Fractionation and partial

characterization of cell walls from normal

and non-ripening mutant tomatoes. Physiologia

Plantarium 62: 25-32.

Hall, L.N., Tucker, G.A. and Smith, C.J.S.

(1993). Antisense inhibition of pectin esterase

gene-expression in transgenic tomatoes.

The Plant

Journal 3: 121-129.

Huber, D.J. (1983). Polyuronide degradation

and hemicellulose modification in ripening

tomato fruit. Journal of the American Society

of Horticultural Science 108: 405-

409.

Kang, I.K., Suh, S.G., Gross, K.C. and

Byun, J.K. (1994). N-terminal amino and

sequence of persimmon fruit β-

galactosidase. Plant Physiology 105: 975-

979.

Koch, J.L. and Nevins, D.J. (1989). Tomato

fruit cell wall. Use of purified tomato polygalacturonase

and pectinmethylesterase

to identify developmental changes in

pectins. Plant Physiology 91: 816-822.

Kitagawa, Y., Kanayama, Y. and Yamaki,

S. (1995). Isolation of β- galactosidase

fractions from Japanese pear: activity

against native cell wall polysaccharides.

Physiologia Plantarum 93: 545-550.

Langley, K.R., Martin, A., Stenning, R.,

Murray, A.J., Hobson, G.E., Schuch,

W.W. and Bird, C.R. (1994). Mechanical

and Optical assessment of the ripening of

tomato fruit with reduced polygalacturonase

activity. Journal of the Science of

Food and Agriculture 66:547-554.

Li, S.C., Han, J.W., Chen, K.C.and Chen

C.S. (2001). Purification and characterization

of isoforms of β-galactosidases in

mung bean seedlings. Phytochemistry 57:

349-359.

Loveland, J., Gutshall, K, Kasmir, J.,

Prema, P. and Brenchley, J. E. (1994).

Charactrization of psychrotropic microorganisms

producing β- galactosidase

activities. Applied and Environmental Microbiology

60: 12 – 18.

Lowry, O.H., Rosebrough, N.J., Farr, A.L.

and Randall, R.J. (1951). Protein measurement

with the Folin phenol reagent.

Journal of Biological Chemistry 193: 265

-275.

Pressey, R. (1983). β-Galactosidases in ripening

tomatoes. Plant Physiology 71: 132-

135.

Ranwala, A.P., Suematsu, C. and Matsuda,

H. (1992). The role of β-galactosidase in

the modification of cell wall components

during muskmelon fruit ripening. Plant

Physiology 100:1318-1325.

Rose, J.K.C. and Bennett, A.B. (1999). Cooperative

disassembly of the cellulosexyloglucan

network of plant cell walls:

parallels between cell expansion and fruit

ripening. Trends in Plant Science 4: 176-

183.

Ross, G.S., Redgwell, R.J. and MacRae,

E.A. (1993). Kiwifruit β-galactosidase:

isolation and activity against specific fruit

cell-wall polysaccharides. Planta. 189:

499-506.

Ross, G.S., Wegrzyn, T., MacRae, E.A.

Redgwell, R.J. (1994). Apple β-

galactosidase: activity against cell wall

polysaccharides and characterization of a

related cDNA clone. Physiologica Plantarum

106: 521-528.

Smith, D.L. and Gross, K.C. (2000). A family

of at least seven β-galactosidase genes

is expressed during tomato fruit development.

Plant Physiology 123: 1173-1183.

Smith, D.L., Abbott, J.A. and Gross, K.C.

(2002). Down-regulation of tomato β-

galactosidase 4 results in decreased fruit

softening. Plant Physiology 129: 1755-

1762.

Tieman, D.M., Harriman, R.W., Ramamohan,

G. and Handa, A.K. (1992). An antisense

pectin methylesterase gene alters

pectin chemistry and soluble solids in to-

mato fruit. The Plant Cell 4: 667-679.

Tieman, D.M. and Handa, A.K. (1994). Reduction

in pectin methylesterase activity

modifies tissue integrity and cation levels

in ripening tomato (Lycopersicon esculentum

Mill.) fruits. Plant Physiology 106:

429- 436.

Wang, T-W., Zhang, C-G., Wu, W.,

Nowack, L.M., Madey, E. and Thompson,

J.E. (2005). Antisense suppression of

deoxyhypusine synthase in tomato delays

fruit softening and alters growth and

development. Plant Physiology 138: 1372-

1382.

Whitaker, J. R. (1990). New and Future use

of enzymes in food processing. Food Biotechnology:

4: 669 – 697.