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Biochemical Characterization of Acyl-CoA: Lysophosphatidylcholine Acyltransferase (LPCAT) Enzyme from the Seeds of Salvia hispanica

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Abstract

Salvia hispanica (chia) is the highest reported terrestrial plant source of alpha-linolenic acid (ALA, ~ 65%), an ω-3 polyunsaturated fatty acid with numerous health benefits. The molecular basis of high ALA accumulation in chia is yet to be understood. We have identified lysophosphatidylcholine acyltransferase (LPCAT) gene from the developing seed transcriptome data of chia and carried out its biochemical characterization through heterologous expression in Saccharomyces cerevisiae. Expression profiling showed that the enzyme was active throughout the seed development, indicating a pivotal role in oil biosynthesis. The enzyme could utilize both saturated and unsaturated lysophosphatidylcholine substrates at the same rate, to synthesize phosphatidylcholine (PC). The enzyme also exhibited lysophosphatidic acid acyltransferase (LPAAT) activity, by preferring lysophosphatidic acid substrate. Substrate specificity studies showed that the enzyme preferred both monounsaturated and polyunsaturated fatty acyl CoAs over saturated CoAs. This activity may play a key role in enriching the PC fraction with polyunsaturated fatty acids (PUFAs). PUFAs present on PC can be transferred to oil through the action of other acyltransferases. Our results describe a new LPCAT enzyme that can be used to biotechnologically alter oilseed crops to incorporate more PUFA in its seed oil.

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Acknowledgements

The authors thank the Director, CSIR-CFTRI, Mysuru for the facilities provided. The research work is financially supported by DST-SERB, under the project (EEQ/2016/000531). Rahul Gopalam is thankful to CSIR-UGC, New Delhi for providing fellowship.

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Authors and Affiliations

  1. Department of Lipid Science, CSIR-Central Food Technological Research Institute (CSIR-CFTRI), Mysore, 570020, India

    Rahul Gopalam, Sharath Bijoor & Ajay W. Tumaney

  2. Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, 201002, India

    Rahul Gopalam & Ajay W. Tumaney

  3. Department of Microbiology and Cell Biology, Indian Institute of Science, Bengaluru, 560012, India

    Akshay Datey & Dipshikha Chakravortty

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  1. Rahul Gopalam
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  2. Akshay Datey
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  3. Sharath Bijoor
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  4. Dipshikha Chakravortty
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  5. Ajay W. Tumaney
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Contributions

RG and AWT planned and designed experiments. RG, AD and SB performed the experiments. RG, DC AWT analyzed the data; RG prepared the original draft; All the authors reviewed, edited and finalized the final manuscript.

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Correspondence to Ajay W. Tumaney.

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Gopalam, R., Datey, A., Bijoor, S. et al. Biochemical Characterization of Acyl-CoA: Lysophosphatidylcholine Acyltransferase (LPCAT) Enzyme from the Seeds of Salvia hispanica. Mol Biotechnol 63, 963–972 (2021). https://doi.org/10.1007/s12033-021-00354-3

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