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Characterization of two acyl-acyl carrier protein thioesterases from developing Cuphea seeds specific for medium-chain- and oleoyl-acyl carrier protein

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Abstract

Two acyl-acyl carrier protein (ACP) thioesterases were partially purified from developing seeds of Cuphea lanceolata Ait., a plant with decanoic acid-rich triacylglycerols. The two enzymes differ markedly in their substrate specificity. One is specific for medium-chain acyl-ACPs, the other one for oleoyl-ACP. In addition, these enzymes are distinct with regard to molecular weight, pH optimum and sensitivity to salt. The thioesterases could be separated by Mono Q chromatography or gel filtration. The medium-chain acyl-ACP thioesterase and oleoyl-ACP thioesterase were purified from a crude extract 29- and 180-fold, respectively. In Cuphea wrightii A. Gray, which predominantly contains decanoic a nd lauric acid in the seeds, two different thioesterases were also found with a similar substrate specificity as in Cuphea lanceolata.

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Abbreviations

ACP:

acyl carrier protein

Ches:

2-[N-cyclo-hexylamino]ethanesulfonic acid

DTT:

dithiothreitol

MCTE:

medium-chain acyl-acyl carrier protein thioesterase

Mes:

2-[N-morpholino]ethanesulfonic acid

OTE:

oleoyl-acyl carrier protein thioesterase; fatty acids are abbreviated as usual in the form: number of carbon atoms in the acyl chain “:” number of double bonds

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Authors and Affiliations

  1. Department of Botany and Plant Pathology, Michigan State University, 48824-1312, East Lansing, MI, USA

    Peter Dörmann & John B. Ohlrogge

  2. Institut für Biochemie, Universität Münster, Wilhelm-Klemm-Strasse 2, W-4400, Münster, FRG

    Peter Dörmann & Friedrich Spener

Authors
  1. Peter Dörmann
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  2. Friedrich Spener
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  3. John B. Ohlrogge
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Additional information

This work was supported by grants of the German Federal Ministry for Research and Technology (Förderkennzeichen 0319412 C), by the German Academic Exchange Service and by the National Science Foundation (grant number DCB 9005290). The authors would like to thank Dr. William Hitz, DuPont, for kindly providing anti-soybean OTE antibodies. Acknowledgement is made to the Michigan Agricultural Experiment Station for its support of this research.

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Dörmann, P., Spener, F. & Ohlrogge, J.B. Characterization of two acyl-acyl carrier protein thioesterases from developing Cuphea seeds specific for medium-chain- and oleoyl-acyl carrier protein. Planta 189, 425–432 (1993). https://doi.org/10.1007/BF00194441

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