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Identification of a cinnamoyl-CoA reductase from Cinnamomum cassia involved in trans-cinnamaldehyde biosynthesis

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Abstract

Main conclusion

The identification of a functional cinnamoyl-CoA reductase enzyme from Cinnamomum cassia involved in trans-cinnamaldehyde biosynthesis offers the potential for enhancing trans-cinnamaldehyde production through genetic engineering.

A significant accumulation of trans-cinnamaldehyde has been found in the bark tissues of C. cassia, used in traditional Chinese medicine. trans-Cinnamaldehyde exhibits various pharmacological properties such as anti-inflammatory, analgesic, and protection of the stomach and the digestive tract. However, further elucidation and characterization of the biosynthetic pathway for trans-cinnamaldehyde is required. In this study, we conducted an integrated analysis of trans-cinnamaldehyde accumulation profiles and transcriptomic data from five different C. cassia tissues to identify the genes involved in its biosynthesis. The transcriptome data we obtained included nearly all genes associated with the trans-cinnamaldehyde pathway, with the majority demonstrating high abundance in branch barks and trunk barks. We successfully cloned four C. cassia cinnamoyl-CoA reductases (CcCCRs), a key gene in trans-cinnamaldehyde biosynthesis. We found that the recombinant CcCCR1 protein was the only one that more efficiently converted cinnamoyl-CoA into trans-cinnamaldehyde. CcCCR1 exhibited approximately 14.7-fold higher catalytic efficiency (kcat/Km) compared to the Arabidopsis thaliana cinnamoyl-CoA reductase 1 (AtCCR1); therefore, it can be utilized for engineering higher trans-cinnamaldehyde production as previously reported. Molecular docking studies and mutagenesis experiments also validated the superior catalytic activity of CcCCR1 compared to AtCCR1. These findings provide valuable insights for the functional characterization of enzyme-coding genes and hold potential for future engineering of trans-cinnamaldehyde biosynthetic pathways.

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Data availability

The CDS and protein sequence of CcCCR1 have been submitted to NCBI GenBank (accession number: OR416486).

Abbreviations

4CL:

4-Coumarate: CoA ligase

CCR:

Cinnamoyl-CoA reductase

PAL:

Phenylalanine ammonia-lyases

t-CA:

trans-Cinnamaldehyde

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Acknowledgement

We are grateful for financial support of the Guangdong Basic and Applied Basic Research Foundation (2020B1515420007) and the Open Competition Program of Ten Major Directions of Agricultural Science and Technology Innovation for the 14th Five-Year Plan of Guangdong Province, China (2022SDZG07) and the Nature Science Foundation of China (32370383).

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Authors and Affiliations

  1. Center for Medicinal Plants Research, College of Life Sciences, South China Agricultural University, Guangzhou, 510642, China

    Peng Ye, Jianmu Su, Jianhao Lin, Yanqun Li & Hong Wu

  2. Guangdong Key Laboratory for Innovative Development and Utilization of Forest Plant Germplasm, College of Forestry, South China Agricultural University, Guangzhou, 510642, China

    Yanqun Li & Hong Wu

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  1. Peng Ye
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  2. Jianmu Su
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  3. Jianhao Lin
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Contributions

Hong Wu and Yanqun Li conceived and designed the research. Peng Ye analyzed data and wrote the manuscript. Jianmu Su and Jianhao Lin assisted during the experiments. All authors read and approved the manuscripts.

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Correspondence to Yanqun Li or Hong Wu.

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Communicated by De-Yu Xie.

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Ye, P., Su, J., Lin, J. et al. Identification of a cinnamoyl-CoA reductase from Cinnamomum cassia involved in trans-cinnamaldehyde biosynthesis. Planta 259, 138 (2024). https://doi.org/10.1007/s00425-024-04419-w

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