Purification and characterization of 1-aminocyclopropane-1-carboxylate N-malonyltransferase from etiolated mung bean hypocotyls

Lining Guo, Richard N. Arteca, Allen T. Phillips, Yu Liu

Research output: Contribution to journalArticle

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Abstract

1-Aminocyclopropane-1-carboxylate (ACC) N-malonyltransferase converts ACC, an immediate precursor of ethylene, to the presumably inactive product malonyl-ACC (MACC). This enzyme plays a role in ethylene production by reducing the level of free ACC in plant tissue. In this study, ACC N-malonyltransferase was purified 3660-fold from etiolated mutig bean (Vigna radiate) hypocotyls, with a 6% overall recovery. The final specific activity was about 83,000 nmol of MACC formed mg-1 protein h-1. The five-step purification protocol consisted of polyethylene glycol fractionation, Cibacron blue 3GA-agarose chromatography using salt gradient elution, Sephadex G-100 gel filtration, MonoQ anion-exchange chromatography, and Cibacron blue 3GA-agarose chromatography using malonyl-CoA plus ACC for elution. The molecular mass of the native enzyme determined by Sephadex G-100 chromatography was 50 ± 3 kD. Protein from the final purification step showed one major band at 55 kD after sodium dodecyl sulfate polyacrylamide gel electrophoresis, indicating that ACC N-malonyltransferase is a monomer. The mung bean ACC N-malonyltransferase has a pH optimum of 8.0, an apparent Km of 0.5 mM for ACC and 0.2 mM for malonyl-coenzyme A, and an Arrhenius activation energy of 70.29 kJ mol-1 degree-1.

Original languageEnglish (US)
Pages (from-to)2041-2045
Number of pages5
JournalPlant physiology
Volume100
Issue number4
DOIs
StatePublished - Dec 1992

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Malonyl Coenzyme A
Hypocotyl
Agarose Chromatography
malonyl coenzyme A
mung beans
hypocotyls
Chromatography
chromatography
agarose
Enzymes
Sodium Dodecyl Sulfate
Gel Chromatography
Anions
Polyacrylamide Gel Electrophoresis
Vigna
Proteins
Salts
1-aminocyclopropane-1-carboxylic acid
enzymes
polyethylene glycol

All Science Journal Classification (ASJC) codes

  • Physiology
  • Genetics
  • Plant Science

Cite this

Guo, Lining ; Arteca, Richard N. ; Phillips, Allen T. ; Liu, Yu. / Purification and characterization of 1-aminocyclopropane-1-carboxylate N-malonyltransferase from etiolated mung bean hypocotyls. In: Plant physiology. 1992 ; Vol. 100, No. 4. pp. 2041-2045.
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abstract = "1-Aminocyclopropane-1-carboxylate (ACC) N-malonyltransferase converts ACC, an immediate precursor of ethylene, to the presumably inactive product malonyl-ACC (MACC). This enzyme plays a role in ethylene production by reducing the level of free ACC in plant tissue. In this study, ACC N-malonyltransferase was purified 3660-fold from etiolated mutig bean (Vigna radiate) hypocotyls, with a 6{\%} overall recovery. The final specific activity was about 83,000 nmol of MACC formed mg-1 protein h-1. The five-step purification protocol consisted of polyethylene glycol fractionation, Cibacron blue 3GA-agarose chromatography using salt gradient elution, Sephadex G-100 gel filtration, MonoQ anion-exchange chromatography, and Cibacron blue 3GA-agarose chromatography using malonyl-CoA plus ACC for elution. The molecular mass of the native enzyme determined by Sephadex G-100 chromatography was 50 ± 3 kD. Protein from the final purification step showed one major band at 55 kD after sodium dodecyl sulfate polyacrylamide gel electrophoresis, indicating that ACC N-malonyltransferase is a monomer. The mung bean ACC N-malonyltransferase has a pH optimum of 8.0, an apparent Km of 0.5 mM for ACC and 0.2 mM for malonyl-coenzyme A, and an Arrhenius activation energy of 70.29 kJ mol-1 degree-1.",
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Purification and characterization of 1-aminocyclopropane-1-carboxylate N-malonyltransferase from etiolated mung bean hypocotyls. / Guo, Lining; Arteca, Richard N.; Phillips, Allen T.; Liu, Yu.

In: Plant physiology, Vol. 100, No. 4, 12.1992, p. 2041-2045.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Purification and characterization of 1-aminocyclopropane-1-carboxylate N-malonyltransferase from etiolated mung bean hypocotyls

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N2 - 1-Aminocyclopropane-1-carboxylate (ACC) N-malonyltransferase converts ACC, an immediate precursor of ethylene, to the presumably inactive product malonyl-ACC (MACC). This enzyme plays a role in ethylene production by reducing the level of free ACC in plant tissue. In this study, ACC N-malonyltransferase was purified 3660-fold from etiolated mutig bean (Vigna radiate) hypocotyls, with a 6% overall recovery. The final specific activity was about 83,000 nmol of MACC formed mg-1 protein h-1. The five-step purification protocol consisted of polyethylene glycol fractionation, Cibacron blue 3GA-agarose chromatography using salt gradient elution, Sephadex G-100 gel filtration, MonoQ anion-exchange chromatography, and Cibacron blue 3GA-agarose chromatography using malonyl-CoA plus ACC for elution. The molecular mass of the native enzyme determined by Sephadex G-100 chromatography was 50 ± 3 kD. Protein from the final purification step showed one major band at 55 kD after sodium dodecyl sulfate polyacrylamide gel electrophoresis, indicating that ACC N-malonyltransferase is a monomer. The mung bean ACC N-malonyltransferase has a pH optimum of 8.0, an apparent Km of 0.5 mM for ACC and 0.2 mM for malonyl-coenzyme A, and an Arrhenius activation energy of 70.29 kJ mol-1 degree-1.

AB - 1-Aminocyclopropane-1-carboxylate (ACC) N-malonyltransferase converts ACC, an immediate precursor of ethylene, to the presumably inactive product malonyl-ACC (MACC). This enzyme plays a role in ethylene production by reducing the level of free ACC in plant tissue. In this study, ACC N-malonyltransferase was purified 3660-fold from etiolated mutig bean (Vigna radiate) hypocotyls, with a 6% overall recovery. The final specific activity was about 83,000 nmol of MACC formed mg-1 protein h-1. The five-step purification protocol consisted of polyethylene glycol fractionation, Cibacron blue 3GA-agarose chromatography using salt gradient elution, Sephadex G-100 gel filtration, MonoQ anion-exchange chromatography, and Cibacron blue 3GA-agarose chromatography using malonyl-CoA plus ACC for elution. The molecular mass of the native enzyme determined by Sephadex G-100 chromatography was 50 ± 3 kD. Protein from the final purification step showed one major band at 55 kD after sodium dodecyl sulfate polyacrylamide gel electrophoresis, indicating that ACC N-malonyltransferase is a monomer. The mung bean ACC N-malonyltransferase has a pH optimum of 8.0, an apparent Km of 0.5 mM for ACC and 0.2 mM for malonyl-coenzyme A, and an Arrhenius activation energy of 70.29 kJ mol-1 degree-1.

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