Avaliação da atividade da lipoxigenase em linhagens de soja

Sawazaki, Haiko Enok; Teixeira, João Paulo Feijão; Miranda, Manoel Albino Coelho de

doi:10.1590/S0006-87051987000200017

Resumos

Com o fim de verificar os melhores métodos para identificação de mutantes de soja para a enzima lipoxigenase e a possível correlação entre atividade enzimática e teor de ácido linolênico, foram analisadas sementes do cultivar IAC-8 e das linhagens A-5 (de baixo teor de ácido linolênico), PI 408251 (mutante menos L1), PI 86023 (mutante menos L2), Tohoko nº 74 (mutante menos L3), e as da geração F2 do cruzamento do 'IAC-8' com as linhagens mutantes (menos L1, menos L2 e menos L3), quanto aos teores de ácido linolênico, atividade de lipoxigenase e ocorrência de genótipos com ausência de isoenzima de lipoxigenase. A quantificação da atividade enzimática foi realizada por método colorimétrico e a detecção das isoenzimas de lipoxigenase, por focalização isoelétrica. O teor de óleo foi determinado gravimetricamente e o de ácidos graxos, por cromatografia em fase gasosa de seus ésteres metílicos. A determinação da atividade enzimática forneceu boa indicação para identificação final das isoenzimas por focalização isoelétrica, permitindo o emprego desse método para a escolha de linhagens em programa de melhoramento genético visando a maior estabilidade do óleo de soja. Para o material estudado, não se encontrou correlação entre atividade enzimática e teor de ácido linolênico.

soja; Glycine max (L) (Merrill); lipoxigenase; ácido linolênico

Seed of soybeans, Glycine max L. (Merrill), cultivar IAC-8 and lines A-5, low linolenic acid content, PI 408251, lacking L1 mutant, PI 86023, lacking L2 mutant, Tohoko nº 74, lacking L3 mutant, and F2 population from the crosses between IAC-8 and mutant lines (lacking L1, L2 and L3) were analysed for linolenic acid content, lipoxygenase activity and identification of mutant lines lacking L1, L2 or L3 isoenzymes. The purpose was to evaluate methods to identify mutants lines of soybean for lipoxygenase and to verify a possible correlation between enzimatic activity and linolenic acid content. The enzimatic activity was estimated quantitatively by a colorimetric method and lipoxygenase isoenzymes detection by isoelectric focusing. The oil seed content was determined by gravimetry and the fatty acids content by gas liquid chromatography of their methyl esters. The lipoxygenase activity data was good to indicate mutant lines for the final identification using isoelectric focusing. It permits the use of this method for choosing lines in breeding program to obtain a good soybean oil stability. It was not observed correlation, in the material studied, between enzymatic activity and linolenic acid content.

soybeans; Glycine max (L.) (Merrill); lipoxygenase; linolenic acid

ARTIGOS

Avaliação da atividade da lipoxigenase em linhagens de soja

Lipoxygenase activity in soybean lines

Haiko Enok Sawazaki^I; João Paulo Feijão TeixeiraI, ^* * Com bolsa de suplementação do CNPq. ; Manoel Albino Coelho de MirandaII, ^* * Com bolsa de suplementação do CNPq.

^ISeção de Fitoquímica, Instituto Agronômico (IAC), Caixa Postal 28, 13001 Campinas (SP)

^IISeção de Leguminosas, IAC

RESUMO

Com o fim de verificar os melhores métodos para identificação de mutantes de soja para a enzima lipoxigenase e a possível correlação entre atividade enzimática e teor de ácido linolênico, foram analisadas sementes do cultivar IAC-8 e das linhagens A-5 (de baixo teor de ácido linolênico), PI 408251 (mutante menos L₁), PI 86023 (mutante menos L₂), Tohoko nº 74 (mutante menos L₃), e as da geração F₂ do cruzamento do 'IAC-8' com as linhagens mutantes (menos L₁, menos L₂ e menos L₃), quanto aos teores de ácido linolênico, atividade de lipoxigenase e ocorrência de genótipos com ausência de isoenzima de lipoxigenase. A quantificação da atividade enzimática foi realizada por método colorimétrico e a detecção das isoenzimas de lipoxigenase, por focalização isoelétrica. O teor de óleo foi determinado gravimetricamente e o de ácidos graxos, por cromatografia em fase gasosa de seus ésteres metílicos. A determinação da atividade enzimática forneceu boa indicação para identificação final das isoenzimas por focalização isoelétrica, permitindo o emprego desse método para a escolha de linhagens em programa de melhoramento genético visando a maior estabilidade do óleo de soja. Para o material estudado, não se encontrou correlação entre atividade enzimática e teor de ácido linolênico.

Termos de indexação: soja, Glycine max (L) (Merrill), lipoxigenase, ácido linolênico.

SUMMARY

Seed of soybeans, Glycine max L. (Merrill), cultivar IAC-8 and lines A-5, low linolenic acid content, PI 408251, lacking L₁ mutant, PI 86023, lacking L₂ mutant, Tohoko nº 74, lacking L₃ mutant, and F₂ population from the crosses between IAC-8 and mutant lines (lacking L₁, L₂ and L₃) were analysed for linolenic acid content, lipoxygenase activity and identification of mutant lines lacking L₁, L₂ or L₃ isoenzymes. The purpose was to evaluate methods to identify mutants lines of soybean for lipoxygenase and to verify a possible correlation between enzimatic activity and linolenic acid content. The enzimatic activity was estimated quantitatively by a colorimetric method and lipoxygenase isoenzymes detection by isoelectric focusing. The oil seed content was determined by gravimetry and the fatty acids content by gas liquid chromatography of their methyl esters. The lipoxygenase activity data was good to indicate mutant lines for the final identification using isoelectric focusing. It permits the use of this method for choosing lines in breeding program to obtain a good soybean oil stability. It was not observed correlation, in the material studied, between enzymatic activity and linolenic acid content.

Index terms: soybeans, Glycine max (L.) (Merrill), lipoxygenase, linolenic acid

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ALLEN, J.C. Soybean lipoxygenase. 1. Purification, and the effect of organic solvents upon the kinetics of the reaction. European Journal Biochemistry, 4:201-208, 1968.
CHIBA, H.; TAKAHASHI, N. & SASAKI, R. Enzymatic improvement of food flavor. II. Removal of beany flavor from soybean products by aldehyde dehydrogenase. Agricultural and Biological Chemistry, 43:1883-1889, 1979.
CHRISTOPHER, J.; PISTORIUS, E. & AXELROB, B. Isolation of an isozyme of soybean lipoxygenase. Biochimica et Biophysica Acta, 198:12-19, 1970.
^__________; PISTORIUS, E.K. & BERNARD, A. Isolation of a third isoenzyme of soybean lipoxygenase. Biochimica et Biophysica Acta, 284:54-62, 1972.
DAVIES, C.S. & NIELSEN, N.C. Genetic analysis of a null-allele for lipoxygenase-2 in soybean. Crop Science, 26:460-463, 1986.
DILLARD, M.G.; HENICK, A.S. & KOCH, R.B. Differences in reactivity of legume lipoxidases. The Journal of Biological Chemistry, 236:37-40, 1961.
FUNK, M.O.; WHITNEY, M.A.; HAUSKENECHT, E.C. & O'BRIEN, E.M. Resolution of the isoenzymes of soybean lipoxygenase using isoelecric focusing and chromatofocusing. Analytical Biochemistry, 146:246-251, 1985.
GUSS, P.L.; RICHARDSON, T. & STAHMANN, M.A. The oxidation-reduction enzymes of wheat. III. Isoenzymes of lipoxidase in wheat fractions and soybean. Cereal Chemistry, 44:607-610, 1967.
GUSS, P.L.; RICHARDSON, T. & STAHMANN, M.A. Oxidation of various lipid substrates with unfractionated soybean and wheat lipoxidase. Journal of the American Oil Chemists Society, 45:272-276, 1968.
HAMMOND, E.G. & FEHR, W.R. Registration of A5 germplast line of soybean. Crop Science, 23:192, 1983.
HARTMAN, L. & LAGO, R.C.A. Rapid preparation of fatty acid methyl esters from lipids. Laboratory Practice, 22:475-477, 1973.
HILDÉBRAND, D.F. & HYMOWITZ, T. Inheritance of lipoxygenase-1 activity in soybean seeds. Crop Science, 22:851-853, 1982.
KITAMURA, K. Biochemical characterization of lipoxygenase lacking mutants, L-1-less, L-2-less, and L-3-less soybean. Agricultural and Biological Chemistry, 48:2339-2346, 1984.
^__________; DAVIES, C.S.; KAIZUMA, N. & NIELSEN, N.C. Genetic analysis of a null-allele for lipoxygenase-3 in soybean seeds. Crop Science, 23:924-927, 1983.
KOCH, R.B. Calcium ion activation of lipoxidase. Archives of Biochemistry and Biophysics, 125:303-307, 1968.
^__________; STERN, B. & FERRARI, C.G. Linoleic acid and trilinolein as substrates for soybean lipoxidase (s). Archives of Biochemistry and Biophysics, 78:165-179, 1958.
LOWRY, O.H.; ROSEBROUGH, N.J.; FARR, A.C. & RANDALL, R.J. Protein measurement with the Folin-phenol reagent. Journal of Biological Chemistry, 193:265-275, 1951.
MATTICK, L.R. & HAND, D.B. Identification of a volatile component in soybean that contributes to the raw bean flavor. Journal of Agricultural and Food Chemistry, 17:15-17, 1969.
MUSTAKAS, G.C.; ALBRECHT, W.J.; McGHEE, J.E.; BLACK, L.T.; BOOKWALTER, G.N. & GRIFFIN JUNIOR, E.L. Lipoxidase deactivation to improve stability, odor and flavor of full-fat soy flours. Journal of the American Oil Chemist's Societ y, 46:623-626, 1969.
RIGHETTI, P.G. & DRYSDALE, J.W. Isoelectric focusing in gels. Journal of Chromatography, 98:271-321, 1974.
SESSA, D.J.; HONIG, D.H. & RACKIS, J.J. Lipid oxidation in full-fat and defatted soybean flakes as related to soybean flavor. Cereal Chemistry, 46:675-686, 1969.
TRIEBOLD, H.O. & AURAND, L.N. Food composition and analysis. New York, Van Nostrand, 1963. 497p.
VERHUE, W.M. & FRANCKE, A. The heterogeneity of soybean lipoxygenase. Biochimica et Biophysica Acta, 284:43-53, 1972.

*

Com bolsa de suplementação do CNPq.

Datas de Publicação

Publicação nesta coleção
06 Dez 2007
Data do Fascículo
1987

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

[1] ALLEN, J.C. Soybean lipoxygenase. 1. Purification, and the effect of organic solvents upon the kinetics of the reaction. European Journal Biochemistry, 4:201-208, 1968.

[2] CHIBA, H.; TAKAHASHI, N. & SASAKI, R. Enzymatic improvement of food flavor. II. Removal of beany flavor from soybean products by aldehyde dehydrogenase. Agricultural and Biological Chemistry, 43:1883-1889, 1979.

[3] CHRISTOPHER, J.; PISTORIUS, E. & AXELROB, B. Isolation of an isozyme of soybean lipoxygenase. Biochimica et Biophysica Acta, 198:12-19, 1970.

[4] ^__________; PISTORIUS, E.K. & BERNARD, A. Isolation of a third isoenzyme of soybean lipoxygenase. Biochimica et Biophysica Acta, 284:54-62, 1972.

[5] DAVIES, C.S. & NIELSEN, N.C. Genetic analysis of a null-allele for lipoxygenase-2 in soybean. Crop Science, 26:460-463, 1986.

[6] DILLARD, M.G.; HENICK, A.S. & KOCH, R.B. Differences in reactivity of legume lipoxidases. The Journal of Biological Chemistry, 236:37-40, 1961.

[7] FUNK, M.O.; WHITNEY, M.A.; HAUSKENECHT, E.C. & O'BRIEN, E.M. Resolution of the isoenzymes of soybean lipoxygenase using isoelecric focusing and chromatofocusing. Analytical Biochemistry, 146:246-251, 1985.

[8] GUSS, P.L.; RICHARDSON, T. & STAHMANN, M.A. The oxidation-reduction enzymes of wheat. III. Isoenzymes of lipoxidase in wheat fractions and soybean. Cereal Chemistry, 44:607-610, 1967.

[9] GUSS, P.L.; RICHARDSON, T. & STAHMANN, M.A. Oxidation of various lipid substrates with unfractionated soybean and wheat lipoxidase. Journal of the American Oil Chemists Society, 45:272-276, 1968.

[10] HAMMOND, E.G. & FEHR, W.R. Registration of A5 germplast line of soybean. Crop Science, 23:192, 1983.

[11] HARTMAN, L. & LAGO, R.C.A. Rapid preparation of fatty acid methyl esters from lipids. Laboratory Practice, 22:475-477, 1973.

[12] HILDÉBRAND, D.F. & HYMOWITZ, T. Inheritance of lipoxygenase-1 activity in soybean seeds. Crop Science, 22:851-853, 1982.

[13] KITAMURA, K. Biochemical characterization of lipoxygenase lacking mutants, L-1-less, L-2-less, and L-3-less soybean. Agricultural and Biological Chemistry, 48:2339-2346, 1984.

[14] ^__________; DAVIES, C.S.; KAIZUMA, N. & NIELSEN, N.C. Genetic analysis of a null-allele for lipoxygenase-3 in soybean seeds. Crop Science, 23:924-927, 1983.

[15] KOCH, R.B. Calcium ion activation of lipoxidase. Archives of Biochemistry and Biophysics, 125:303-307, 1968.

[16] ^__________; STERN, B. & FERRARI, C.G. Linoleic acid and trilinolein as substrates for soybean lipoxidase (s). Archives of Biochemistry and Biophysics, 78:165-179, 1958.

[17] LOWRY, O.H.; ROSEBROUGH, N.J.; FARR, A.C. & RANDALL, R.J. Protein measurement with the Folin-phenol reagent. Journal of Biological Chemistry, 193:265-275, 1951.

[18] MATTICK, L.R. & HAND, D.B. Identification of a volatile component in soybean that contributes to the raw bean flavor. Journal of Agricultural and Food Chemistry, 17:15-17, 1969.

[19] MUSTAKAS, G.C.; ALBRECHT, W.J.; McGHEE, J.E.; BLACK, L.T.; BOOKWALTER, G.N. & GRIFFIN JUNIOR, E.L. Lipoxidase deactivation to improve stability, odor and flavor of full-fat soy flours. Journal of the American Oil Chemist's Societ y, 46:623-626, 1969.

[20] RIGHETTI, P.G. & DRYSDALE, J.W. Isoelectric focusing in gels. Journal of Chromatography, 98:271-321, 1974.

[21] SESSA, D.J.; HONIG, D.H. & RACKIS, J.J. Lipid oxidation in full-fat and defatted soybean flakes as related to soybean flavor. Cereal Chemistry, 46:675-686, 1969.

[22] TRIEBOLD, H.O. & AURAND, L.N. Food composition and analysis. New York, Van Nostrand, 1963. 497p.

[23] VERHUE, W.M. & FRANCKE, A. The heterogeneity of soybean lipoxygenase. Biochimica et Biophysica Acta, 284:43-53, 1972.

Brasil

Brasil

Avaliação da atividade da lipoxigenase em linhagens de soja

Lipoxygenase activity in soybean lines

Resumos

Datas de Publicação