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Protocol: An updated integrated methodology for analysis of metabolites and enzyme activities of ethylene biosynthesis.

Bulens I, Van de Poel B, Hertog ML, De Proft MP, Geeraerd AH, Nicolaï BM - Plant Methods (2011)

Bottom Line: The foundations for ethylene research were laid many years ago by researchers such as Lizada, Yang and Hoffman.Technological developments since then have led to small but significant improvements, contributing to a more efficient workflow.The detailed protocol allows other scientists to rapidly implement these methods in their own laboratories in a consistent and efficient way.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Mechatronics, Biostatistics and Sensors (MeBioS), Department of Biosystems (BIOSYST), Katholieke Universiteit Leuven, Willem de Croylaan 42, bus 2428, B-3001 Leuven, Belgium. inge.bulens@biw.kuleuven.be.

ABSTRACT

Background: The foundations for ethylene research were laid many years ago by researchers such as Lizada, Yang and Hoffman. Nowadays, most of the methods developed by them are still being used. Technological developments since then have led to small but significant improvements, contributing to a more efficient workflow. Despite this, many of these improvements have never been properly documented.

Results: This article provides an updated, integrated set of protocols suitable for the assembly of a complete picture of ethylene biosynthesis, including the measurement of ethylene itself. The original protocols for the metabolites 1-aminocyclopropane-1-carboxylic acid and 1-(malonylamino)cyclopropane-1-carboxylic acid have been updated and downscaled, while protocols to determine in vitro activities of the key enzymes 1-aminocyclopropane-1-carboxylate synthase and 1-aminocyclopropane-1-carboxylate oxidase have been optimised for efficiency, repeatability and accuracy. All the protocols described were optimised for apple fruit, but have been proven to be suitable for the analysis of tomato fruit as well.

Conclusions: This work collates an integrated set of detailed protocols for the measurement of components of the ethylene biosynthetic pathway, starting from well-established methods. These protocols have been optimised for smaller sample volumes, increased efficiency, repeatability and accuracy. The detailed protocol allows other scientists to rapidly implement these methods in their own laboratories in a consistent and efficient way.

No MeSH data available.


Pathway of the ethylene biosynthesis. The ethylene biosynthesis starts from the conversion of S-adenosyl-L-methione (SAM), into 1-aminocyclopropane-1-carboxylic acid (ACC) by the enzyme 1-aminocyclopropane-1-carboxylate synthase (ACS). ACC can then be converted to 1-(malonylamino)cyclopropane-1-carboxylic acid (MACC), by ACC N-Malonyl transferase, or to the end product ethylene, by 1-aminocyclopropane-1-carboxylate oxidase (ACO).
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Figure 1: Pathway of the ethylene biosynthesis. The ethylene biosynthesis starts from the conversion of S-adenosyl-L-methione (SAM), into 1-aminocyclopropane-1-carboxylic acid (ACC) by the enzyme 1-aminocyclopropane-1-carboxylate synthase (ACS). ACC can then be converted to 1-(malonylamino)cyclopropane-1-carboxylic acid (MACC), by ACC N-Malonyl transferase, or to the end product ethylene, by 1-aminocyclopropane-1-carboxylate oxidase (ACO).

Mentions: Ethylene biosynthesis starts from the conversion of S-adenosyl-L-methione (SAM) into 1-aminocyclopropane-1-carboxylic acid (ACC) by the enzyme 1-aminocyclopropane-1-carboxylate synthase (ACS). ACC can then be converted to either 1-(malonylamino)cyclopropane-1-carboxylic acid (MACC) by ACC N-Malonyl transferase, or to the end product, ethylene, by 1-aminocyclopropane-1-carboxylate oxidase (ACO) [1] (Figure 1).


Protocol: An updated integrated methodology for analysis of metabolites and enzyme activities of ethylene biosynthesis.

Bulens I, Van de Poel B, Hertog ML, De Proft MP, Geeraerd AH, Nicolaï BM - Plant Methods (2011)

Pathway of the ethylene biosynthesis. The ethylene biosynthesis starts from the conversion of S-adenosyl-L-methione (SAM), into 1-aminocyclopropane-1-carboxylic acid (ACC) by the enzyme 1-aminocyclopropane-1-carboxylate synthase (ACS). ACC can then be converted to 1-(malonylamino)cyclopropane-1-carboxylic acid (MACC), by ACC N-Malonyl transferase, or to the end product ethylene, by 1-aminocyclopropane-1-carboxylate oxidase (ACO).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3142538&req=5

Figure 1: Pathway of the ethylene biosynthesis. The ethylene biosynthesis starts from the conversion of S-adenosyl-L-methione (SAM), into 1-aminocyclopropane-1-carboxylic acid (ACC) by the enzyme 1-aminocyclopropane-1-carboxylate synthase (ACS). ACC can then be converted to 1-(malonylamino)cyclopropane-1-carboxylic acid (MACC), by ACC N-Malonyl transferase, or to the end product ethylene, by 1-aminocyclopropane-1-carboxylate oxidase (ACO).
Mentions: Ethylene biosynthesis starts from the conversion of S-adenosyl-L-methione (SAM) into 1-aminocyclopropane-1-carboxylic acid (ACC) by the enzyme 1-aminocyclopropane-1-carboxylate synthase (ACS). ACC can then be converted to either 1-(malonylamino)cyclopropane-1-carboxylic acid (MACC) by ACC N-Malonyl transferase, or to the end product, ethylene, by 1-aminocyclopropane-1-carboxylate oxidase (ACO) [1] (Figure 1).

Bottom Line: The foundations for ethylene research were laid many years ago by researchers such as Lizada, Yang and Hoffman.Technological developments since then have led to small but significant improvements, contributing to a more efficient workflow.The detailed protocol allows other scientists to rapidly implement these methods in their own laboratories in a consistent and efficient way.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Mechatronics, Biostatistics and Sensors (MeBioS), Department of Biosystems (BIOSYST), Katholieke Universiteit Leuven, Willem de Croylaan 42, bus 2428, B-3001 Leuven, Belgium. inge.bulens@biw.kuleuven.be.

ABSTRACT

Background: The foundations for ethylene research were laid many years ago by researchers such as Lizada, Yang and Hoffman. Nowadays, most of the methods developed by them are still being used. Technological developments since then have led to small but significant improvements, contributing to a more efficient workflow. Despite this, many of these improvements have never been properly documented.

Results: This article provides an updated, integrated set of protocols suitable for the assembly of a complete picture of ethylene biosynthesis, including the measurement of ethylene itself. The original protocols for the metabolites 1-aminocyclopropane-1-carboxylic acid and 1-(malonylamino)cyclopropane-1-carboxylic acid have been updated and downscaled, while protocols to determine in vitro activities of the key enzymes 1-aminocyclopropane-1-carboxylate synthase and 1-aminocyclopropane-1-carboxylate oxidase have been optimised for efficiency, repeatability and accuracy. All the protocols described were optimised for apple fruit, but have been proven to be suitable for the analysis of tomato fruit as well.

Conclusions: This work collates an integrated set of detailed protocols for the measurement of components of the ethylene biosynthetic pathway, starting from well-established methods. These protocols have been optimised for smaller sample volumes, increased efficiency, repeatability and accuracy. The detailed protocol allows other scientists to rapidly implement these methods in their own laboratories in a consistent and efficient way.

No MeSH data available.