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Biochemical and Catalytic Properties of Three Recombinant Alcohol Acyltransferases of Melon. Sulfur-Containing Ester Formation, Regulatory Role of CoA-SH in Activity, and Sequence Elements Conferring Substrate Preference

Lucchetta, Luciano and Manriquez, Daniel and El-Sharkawy, Islam and Flores, Francisco B. and Sanchez-Bel, Paloma and Zouine, Mohamed and Ginies, Christian and Bouzayen, Mondher and Rombaldi, César and Pech, Jean-Claude and Latché, Alain Biochemical and Catalytic Properties of Three Recombinant Alcohol Acyltransferases of Melon. Sulfur-Containing Ester Formation, Regulatory Role of CoA-SH in Activity, and Sequence Elements Conferring Substrate Preference. (2007) Journal of Agricultural and Food Chemistry, vol. 5 . pp. 5213-5220. ISSN 0021-8561

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Official URL: http://dx.doi.org/10.1021/jf070210w

Abstract

Alcohol acyltransferases (AAT) play a key role in the biosynthesis of ester aroma volatiles in fruit. Three ripening-specific recombinant AATs of cantaloupe Charentais melon fruit (Cm-AAT1, Cm- AAT3, and Cm-AAT4) are capable of synthesizing thioether esters with Cm-AAT1 being by far the most active. All proteins, as well as AAT(s) extracted from melon fruit, are active as tetramers of around 200 kDa. Kinetic analysis demonstrated that CoA-SH, a product of the reaction, is an activator at low concentrations and an inhibitor at higher concentrations. This was confirmed by the addition of phosphotransacetylase at various concentrations, capable of modulating the level of CoA-SH in the reaction medium. Site-directed mutagenesis of some amino acids that were specific to the Cm- AAT sequences into amino acids that were consensus to other characterized AATs greatly affected the selectivity of the original protein and the number of esters produced.

Item Type:Article
Additional Information:The definitive version is available at http://pubs.acs.org The original PDF of the article can be found at Journal of Agricultural and Food Chemistry website : http://pubs.acs.org/doi/abs/10.1021/jf070210w
Audience (journal):International peer-reviewed journal
Uncontrolled Keywords:
Institution:French research institutions > Institut National de la Recherche Agronomique - INRA
Université de Toulouse > Institut National Polytechnique de Toulouse - INPT
Other partners > Universidade Federal de Pelotas - UFPel (BRAZIL)
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Deposited By:Dabernat Séverine

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