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Identification and functional characterization of two Δ12-fatty acid desaturases associated with essential linoleic acid biosynthesis in Physcomitrella patens

  • Genetics and Molecular Biology of Industrial Organisms
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Two Δ12-desaturases associated with the primary steps of long-chain polyunsaturated fatty acid (LC-PUFA) biosynthesis were successfully cloned from Physcomitrella patens and their functions identified. The open reading frames (ORFs) of PpFAD2-1 and PpFAD2-2 consisted of 1,128 bp and code for 375 amino acids. Their deduced polypeptides showed 62–64 % identity to microsomal Δ12-desaturases from other higher plants, and each contained the three histidine clusters typical of the catalytic domains of such enzymes. Yeast cells transformed with plasmid constructs containing PpFAD2-1 or PpFAD2-2 produced an appreciable amount of hexadecadienoic (16:2 Δ9,12) and linoleic acids (18:2 Δ9,12), not normally present in wild-type yeast cells, indicating that the genes encoded functional Δ12-desaturase enzymes. In addition, reduction of the growth temperature from 30 to 15 °C resulted in increased accumulation of unsaturated fatty acid products.

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Acknowledgments

This research was supported by Prince of Songkla University (PSU, PHA540648S), the Higher Education Research Promotion and National Research University Project of Thailand, Office of the Higher Education Commission (PHA5405395), and the Marine Natural Products Research Unit (MNP) at the Faculty of Pharmaceutical Sciences, PSU. We gratefully acknowledge Assist. Prof. Dr. Akkharawit Kanjana-Opas for providing the laboratory facilities and also thank Dr. Brian Hodgson of PSU for assistance with the English.

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Authors and Affiliations

  1. Marine Natural Products Research Unit, Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Prince of Songkla University, Songkhla, 90112, Thailand

    Pichit Chodok & Sireewan Kaewsuwan

  2. Faculty of Science and Technology, Phranakorn si Ayutthaya Rajabhat University, Phranakorn si Ayutthaya, 13000, Thailand

    Pradinunt Eiamsa-ard

  3. Department of Biology, Washington University in St. Louis, St. Louis, MO, 63130-4899, USA

    David J. Cove & Ralph S. Quatrano

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  1. Pichit Chodok
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  2. Pradinunt Eiamsa-ard
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  3. David J. Cove
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  5. Sireewan Kaewsuwan
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Correspondence to Sireewan Kaewsuwan.

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Chodok, P., Eiamsa-ard, P., Cove, D.J. et al. Identification and functional characterization of two Δ12-fatty acid desaturases associated with essential linoleic acid biosynthesis in Physcomitrella patens . J Ind Microbiol Biotechnol 40, 901–913 (2013). https://doi.org/10.1007/s10295-013-1285-3

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