Detection of 1-aminocyclopropane-1-carboxylate oxidase activity in seeds of Stylosanthes humilis H.B.K

Barros, Raimundo Santos; Pinheiro, Frank James Araújo; Müller, Caroline; Pires, Marcel Viana; Silva, Alexei Gresta Vieira da; Ribeiro, Dimas Mendes

Abstract

The activity of 1-aminocyclopropane-1-carboxylate oxidase (ACO) was characterized in seeds of the tropical legume Townsville stylo (Stylosanthes humilis) both in vitro (desalted extract of non-dormant seeds) and in vivo (entire dormant seeds). Optimum conditions for maximum in vitro ACO activity in a Trizma-HCl 100 mM buffered medium were: pH 7.0, temperature 32ºC and cofactors and co-substrate at the following concentrations: NaHCO3 30 mM, sodium ascorbate 30 mM and FeSO4 50 µM. Rates of in vitro reaction catalyzed by ACO were shown to be constant within the interval 15-150 minutes from the onset of the reaction. The apparent Km for in vitro ACO, as determined from the non-linear curve fitting to the Michaelis-Menten equation, was 156±8.3 µM ACC with a Vmax 5.4±0.08 mmol (ET) g-1 h-1 on a fresh matter (FM) basis. In vivo (control basal medium: HCl-KOH 10 mM pH 7.0, 30ºC, reaction time 15 hours) apparent Km was 230±27 µM ACC and Vmax 11.9±0.38 mmol (ET) g-1.h-1 on a FM basis. These data suggest that the enzyme exhibits a relatively low affinity for the substrate. The well-known inhibitors of ACO activity, α-aminoisobutyric acid, salicylic and acetylsalicylic acids, n-propylgallate and cobaltous ions, were highly effective in inhibiting ACO activity of Townsville stylo seeds.

dioxygenase; enzymatic kinetics; ethylene; inhibition; ACO

References

Barros RS, Delatorre CA (1998) 1-Aminocyclopropane-1-carboxylic acid-stimulated germination of dormant seeds of Stylosanthes humilis is inhibited by abscisic acid. Seed Science and Technology 26:319-324.
Binnie JE, McManus MT (2009) Characterization of the 1-aminocyclopropane-1-1carboxylic acid (ACC) oxidase multigene family of Malus domestica Borkh. Phytochemistry 70:348-360.
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248-254.
Calvo AP, Nicolás C, Lorenzo O, Nicolás G, Rodríguez D (2004) Evidence for positive regulation by gibberellins and ethylene of ACC oxidase expression and activity during transition from dormancy to germination in Fagus sylvatica L. seeds. Journal of Plant Growth Regulation 23:44-53.
De Rueda PM, Gallardo M, Matilla AJ, Sánchez-Calle IM (1995) Preliminary characterization of 1-aminocyclopropane-1-carboxylate oxidase properties from embryonic axes of chick-pea (Circer arietinum L.) seeds. Journal of Experimental Botany 46:695-700.
Dilley DR, Kuai J, Poneleit L, Zhu Y, Pekker Y, Wilson ID, Burmeister DM, Gran C, Bowers A (1993) Purification and characterization of ACC oxidase and its expression during ripening in apple fruit. In: Peach JC, Latché A, Balagué C (eds) Cellular and molecular aspects of the plant hormone ethylene, pp. 46-52. Kluwer Academic Publishers, Dordrecht, The Netherlands.
Dunkley HM, Golden KD (1998) ACC oxidase from Carica papaya: isolation and characterization. Physiologia Plantarum 103:225-232.
Dupille E, Zacarías L (1996) Extraction and biochemical characterization of wound-induced ACC oxidase from Citrus peel. Plant Science 114:53-60.
Fan X, Mattheis JP, Fellman JK (1996) Inhibition of apple fruit 1-aminocyclopropane-1-carboxylic acid oxidase activity and respiration by acetylsalicylic acid. Journal of Plant Physiology 140:469-471.
Fernández-Maculet JC, Dong JG, Yang SF (1993) Activation of 1-aminocyclopropane-1-carboxylate oxidase by carbon dioxide. Biochemical and Biophysical Communications 193:1168-1173.
Fernández-Maculet JC, Yang SF (1992) Extraction and partial characterization of the ethylene-forming enzyme from apple fruit. Plant Physiology 99:751-754.
Gómez-Jiménez MC, Matilla AJ, Garrido D (1998) Isolation and characterization of a cDNA encoding an ACC oxidase from Cicer arietinum and its expression during embryogenesis and germination. Australian Journal of Plant Physiology 25:765-773.
Hermann K, Meinhard J, Dobrev P, Linkies A, Pesek B, Hess B, Macháckova I, Fisher U, Leubner-Metzger G (2007) 1-Aminocyclopropane-1-carboxylic acid and abscisic acid during the germination of sugar beet (Beta vulgaris L.): a comparative study of fruits and seeds. Journal of Experimental Botany 58:3047-3060.
John P (1997) Ethylene biosynthesis: the role of 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase, and its possible evolutionary origin. Physiologia Plantarum 100:583-592.
Kepczynski J, Kepczynska E (1997) Ethylene in seed dormancy and germination. Physiologia Plantarum 101:720-726.
Kruzname D, Ievinsh G (1999) Characterization of 1-aminocyclopropane-1-carboxylic acid oxidase from barley (Hordeum vulgare L.) seedlings and pine (Pinus sylvestris L) needles. Plant Science 142:13-19.
Linkies A, Müller K, Morris K, Tureckova V, Wenk M, Cadman CS, Corbineau F, Strnad M, Lyn JR, Finch-Savage WE, Leubner-Metzger G (2009) Ethylene interacts with abscisic acid to regulate endosperm rupture during germination: a comparative approach using Lepidium sativum and Arabidopsis thaliana The Plant Cell 21:3803-3822.
McGarvey DJ, Christoffersen RE (1992) Characterization and kinetic parameters of ethylene-forming enzyme from avocado fruit. Journal of Biological Chemistry 267:5964-6967.
Mohamed AH, Ejeta G, Housley TL (2001) Striga asiatica seed conditioning and 1-aminocyclopropane-1-carboxylate oxidase activity. Weed Research 41:165-176.
Moya-Léon M, John P (1995) Purification and biochemical characterization of 1-aminocyclopropane-1-carboxylate oxidase from banana fruit. Phytochemistry 39:15-20.
Nijenhus-De Vries MA, Woltering EJ, Vrije T (1994) Partial characterization of petal 1-aminocyclopropane-1-carboxylate oxidase. Journal of Plant Physiology 144: 549-554.
Osborne DJ, Walters J, Milborrow BV, Norville A, Stange LMC (1996) Evidence for a non ACC ethylene biosynthesis pathway in lower plants. Phytochemistry 42:51-60.
Pech JC, Latché A, Bouzayen M (2010) Ethylene biosynthesis. In: Davies P (ed) Plant hormones: biosynthesis, signal transduction, action! 3nd, pp. 115-136. Springer Dordrecht, The Netherlands.
Petruzelli L, Sturaro M, Manieri D, Leubner-Metzger G (2003) Calcium requirement for ethylene-dependent response involving 1-aminocyclopropane-1-carboxylic acid in radicle tissues of germinated pea seeds. Plant Cell and Environmental 26:661-667.
Ribeiro DM, Barros RS (2006) Sensitivity to ethylene as a major component in the germination of seeds of Stylosanthes humilis Seed Science Research 16:37-45.
Ritchie RJ, Prvan T (1996) Current statistical methods for estimating the K_m and V_maxof Michaelis-Menten kinetics. Biochemical Education 24:196-206.
Rodríguez-Gacio MC, Matilla AJ (2001) The last step of the ethylene-biosynthesis pathway in turnip tops (Brassica rapa) seed: alterations related to development and germination and its inhibition during desiccation. Physiologia Plantarum 11:273-279.
Rodríguez-Gacio MC, Nicolas C, Matilla AJ (2004) The final step of ethylene biosynthesis pathway in turnip tops (Brassica rapa): molecular characterization of the 1-aminocyclopropane-1-carboxilate oxidase BrACO1 throughout zygotic embryogenesis and germination of heterogeneous seeds. Physiologia Plantarum 121:132-140.
Rudus I, Sasiak M, Kepczynski J (2013) Regulation of ethylene biosynthesis at the level of 1-aminocyclopropane-1-carboxylte oxidase (ACO) gene. Acta Physiologiae Plantarum 35:295-307.
Satoh S, Esashi Y (1983) α-Aminoisobutyric acid, propyl gallate and cobalt ion and the mode of inhibition of ethylene production by cotyledonary segments of cocklebur seeds. Physiologia Plantarum 57:521-526.
Schnell S (2003) A century of enzyme kinetics: reliability of the K_m and V_max estimates. Comments on Theoretical Biology 8:169-187.
Sjörgen E, Lennermas H, Andersson TB, Gråsjö J, Bredberg U (2009) The multiple depletion curves method provides accurate estimates of intrinsic clearance (C_Lint), maximum velocity of the metabolic reaction (V_max) and Michaelis constant (K_m): accuracy and robustness evaluated through experimental data and Monte Carlo simulations. Drug Metabolism and Disposition 37:47-58.
Smith JJ, John P (1993) A activation of 1-aminocyclopropane- 1-carboxylate oxidase by bicarbonate/ carbon dioxide. Phytochemistry 32:1381-1386.
Smith JJ, Ververidis P, John P (1992) Characterization of the ethylene-forming enzyme partially purified from melon. Phytochemistry 31:1485-1494.
Ververidis P, John P (1991) Complete recovery in vitro of ethylene-forming enzyme activity. Phytochemistry 30:725-727.
Vioque B, Castellano JM (1994) Extraction and biochemical characterization of 1-aminocyclopropane-1-carboxilic acid oxidase from pear. Physiologia Plantarum 90:334-338.
Williams OJ, Golden KD (2002) Purification and characterization of ACC oxidase from Artocarpus altilis Plant Physiology and Biochemistry 40:273-279.
Yang SF, Hoffmann NE (1984) Ethylene biosynthesis and its regulation in higher plants. Annual Review of Plant Physiology 35:155-189.

Publication Dates

Publication in this collection
02 Dec 2013
Date of issue
2013

History

Received
29 Aug 2013
Accepted
16 Oct 2013

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

[1] Barros RS, Delatorre CA (1998) 1-Aminocyclopropane-1-carboxylic acid-stimulated germination of dormant seeds of Stylosanthes humilis is inhibited by abscisic acid. Seed Science and Technology 26:319-324.

[2] Binnie JE, McManus MT (2009) Characterization of the 1-aminocyclopropane-1-1carboxylic acid (ACC) oxidase multigene family of Malus domestica Borkh. Phytochemistry 70:348-360.

[3] Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248-254.

[4] Calvo AP, Nicolás C, Lorenzo O, Nicolás G, Rodríguez D (2004) Evidence for positive regulation by gibberellins and ethylene of ACC oxidase expression and activity during transition from dormancy to germination in Fagus sylvatica L. seeds. Journal of Plant Growth Regulation 23:44-53.

[5] De Rueda PM, Gallardo M, Matilla AJ, Sánchez-Calle IM (1995) Preliminary characterization of 1-aminocyclopropane-1-carboxylate oxidase properties from embryonic axes of chick-pea (Circer arietinum L.) seeds. Journal of Experimental Botany 46:695-700.

[6] Dilley DR, Kuai J, Poneleit L, Zhu Y, Pekker Y, Wilson ID, Burmeister DM, Gran C, Bowers A (1993) Purification and characterization of ACC oxidase and its expression during ripening in apple fruit. In: Peach JC, Latché A, Balagué C (eds) Cellular and molecular aspects of the plant hormone ethylene, pp. 46-52. Kluwer Academic Publishers, Dordrecht, The Netherlands.

[7] Dunkley HM, Golden KD (1998) ACC oxidase from Carica papaya: isolation and characterization. Physiologia Plantarum 103:225-232.

[8] Dupille E, Zacarías L (1996) Extraction and biochemical characterization of wound-induced ACC oxidase from Citrus peel. Plant Science 114:53-60.

[9] Fan X, Mattheis JP, Fellman JK (1996) Inhibition of apple fruit 1-aminocyclopropane-1-carboxylic acid oxidase activity and respiration by acetylsalicylic acid. Journal of Plant Physiology 140:469-471.

[10] Fernández-Maculet JC, Dong JG, Yang SF (1993) Activation of 1-aminocyclopropane-1-carboxylate oxidase by carbon dioxide. Biochemical and Biophysical Communications 193:1168-1173.

[11] Fernández-Maculet JC, Yang SF (1992) Extraction and partial characterization of the ethylene-forming enzyme from apple fruit. Plant Physiology 99:751-754.

[12] Gómez-Jiménez MC, Matilla AJ, Garrido D (1998) Isolation and characterization of a cDNA encoding an ACC oxidase from Cicer arietinum and its expression during embryogenesis and germination. Australian Journal of Plant Physiology 25:765-773.

[13] Hermann K, Meinhard J, Dobrev P, Linkies A, Pesek B, Hess B, Macháckova I, Fisher U, Leubner-Metzger G (2007) 1-Aminocyclopropane-1-carboxylic acid and abscisic acid during the germination of sugar beet (Beta vulgaris L.): a comparative study of fruits and seeds. Journal of Experimental Botany 58:3047-3060.

[14] John P (1997) Ethylene biosynthesis: the role of 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase, and its possible evolutionary origin. Physiologia Plantarum 100:583-592.

[15] Kepczynski J, Kepczynska E (1997) Ethylene in seed dormancy and germination. Physiologia Plantarum 101:720-726.

[16] Kruzname D, Ievinsh G (1999) Characterization of 1-aminocyclopropane-1-carboxylic acid oxidase from barley (Hordeum vulgare L.) seedlings and pine (Pinus sylvestris L) needles. Plant Science 142:13-19.

[17] Linkies A, Müller K, Morris K, Tureckova V, Wenk M, Cadman CS, Corbineau F, Strnad M, Lyn JR, Finch-Savage WE, Leubner-Metzger G (2009) Ethylene interacts with abscisic acid to regulate endosperm rupture during germination: a comparative approach using Lepidium sativum and Arabidopsis thaliana The Plant Cell 21:3803-3822.

[18] McGarvey DJ, Christoffersen RE (1992) Characterization and kinetic parameters of ethylene-forming enzyme from avocado fruit. Journal of Biological Chemistry 267:5964-6967.

[19] Mohamed AH, Ejeta G, Housley TL (2001) Striga asiatica seed conditioning and 1-aminocyclopropane-1-carboxylate oxidase activity. Weed Research 41:165-176.

[20] Moya-Léon M, John P (1995) Purification and biochemical characterization of 1-aminocyclopropane-1-carboxylate oxidase from banana fruit. Phytochemistry 39:15-20.

[21] Nijenhus-De Vries MA, Woltering EJ, Vrije T (1994) Partial characterization of petal 1-aminocyclopropane-1-carboxylate oxidase. Journal of Plant Physiology 144: 549-554.

[22] Osborne DJ, Walters J, Milborrow BV, Norville A, Stange LMC (1996) Evidence for a non ACC ethylene biosynthesis pathway in lower plants. Phytochemistry 42:51-60.

[23] Pech JC, Latché A, Bouzayen M (2010) Ethylene biosynthesis. In: Davies P (ed) Plant hormones: biosynthesis, signal transduction, action! 3nd, pp. 115-136. Springer Dordrecht, The Netherlands.

[24] Petruzelli L, Sturaro M, Manieri D, Leubner-Metzger G (2003) Calcium requirement for ethylene-dependent response involving 1-aminocyclopropane-1-carboxylic acid in radicle tissues of germinated pea seeds. Plant Cell and Environmental 26:661-667.

[25] Ribeiro DM, Barros RS (2006) Sensitivity to ethylene as a major component in the germination of seeds of Stylosanthes humilis Seed Science Research 16:37-45.

[26] Ritchie RJ, Prvan T (1996) Current statistical methods for estimating the K_m and V_maxof Michaelis-Menten kinetics. Biochemical Education 24:196-206.

[27] Rodríguez-Gacio MC, Matilla AJ (2001) The last step of the ethylene-biosynthesis pathway in turnip tops (Brassica rapa) seed: alterations related to development and germination and its inhibition during desiccation. Physiologia Plantarum 11:273-279.

[28] Rodríguez-Gacio MC, Nicolas C, Matilla AJ (2004) The final step of ethylene biosynthesis pathway in turnip tops (Brassica rapa): molecular characterization of the 1-aminocyclopropane-1-carboxilate oxidase BrACO1 throughout zygotic embryogenesis and germination of heterogeneous seeds. Physiologia Plantarum 121:132-140.

[29] Rudus I, Sasiak M, Kepczynski J (2013) Regulation of ethylene biosynthesis at the level of 1-aminocyclopropane-1-carboxylte oxidase (ACO) gene. Acta Physiologiae Plantarum 35:295-307.

[31] Satoh S, Esashi Y (1983) α-Aminoisobutyric acid, propyl gallate and cobalt ion and the mode of inhibition of ethylene production by cotyledonary segments of cocklebur seeds. Physiologia Plantarum 57:521-526.

[32] Schnell S (2003) A century of enzyme kinetics: reliability of the K_m and V_max estimates. Comments on Theoretical Biology 8:169-187.

[33] Sjörgen E, Lennermas H, Andersson TB, Gråsjö J, Bredberg U (2009) The multiple depletion curves method provides accurate estimates of intrinsic clearance (C_Lint), maximum velocity of the metabolic reaction (V_max) and Michaelis constant (K_m): accuracy and robustness evaluated through experimental data and Monte Carlo simulations. Drug Metabolism and Disposition 37:47-58.

[34] Smith JJ, John P (1993) A activation of 1-aminocyclopropane- 1-carboxylate oxidase by bicarbonate/ carbon dioxide. Phytochemistry 32:1381-1386.

[35] Smith JJ, Ververidis P, John P (1992) Characterization of the ethylene-forming enzyme partially purified from melon. Phytochemistry 31:1485-1494.

[36] Ververidis P, John P (1991) Complete recovery in vitro of ethylene-forming enzyme activity. Phytochemistry 30:725-727.

[37] Vioque B, Castellano JM (1994) Extraction and biochemical characterization of 1-aminocyclopropane-1-carboxilic acid oxidase from pear. Physiologia Plantarum 90:334-338.

[38] Williams OJ, Golden KD (2002) Purification and characterization of ACC oxidase from Artocarpus altilis Plant Physiology and Biochemistry 40:273-279.

[39] Yang SF, Hoffmann NE (1984) Ethylene biosynthesis and its regulation in higher plants. Annual Review of Plant Physiology 35:155-189.