Molecular cloning and characterisation of two enzymes involved in the rosmarinic acid biosynthesis pathway of Prunella vulgaris L.

详细信息    查看全文

• 作者：Mei Ru ; Kunru Wang ; Zhenqing Bai ; Liang Peng…
• 关键词：Prunella vulgaris ; Rosmarinic acid ; Rosmarinic acid synthase ; Cytochrome P450 monooxygenase ; Subcellular localisation ; Expression pattern
• 刊名：Plant Cell, Tissue and Organ Culture (PCTOC)
• 出版年：2017
• 出版时间：February 2017
• 年：2017
• 卷：128
• 期：2
• 页码：381-390
• 全文大小：
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Plant Sciences; Plant Physiology; Plant Genetics & Genomics; Plant Pathology;
• 出版者：Springer Netherlands
• ISSN：1573-5044
• 卷排序：128

文摘

Prunella vulgaris, a widely-used medicinal perennial herb, contains various active compounds and has multifaceted medicinal activities. As the index component in P. vulgaris, rosmarinic acid (RA) is a typical phenolic acid, with significant anti-inflammatory, anti-oxidant, anti-tumour, anti-viral, and anti-microbial properties. To better understand the RA biosynthetic pathway in P. vulgaris, we isolated and cloned the cDNA sequences of putative RA synthase (PvRAS) and cytochrome P450 monooxygenase (PvCYP98A101), two important enzymes catalysing RA biosynthesis. Sequence analysis revealed that PvRAS contained an open reading frame (ORF) of 1305 bp encoding a 435-amino-acid residue belonging to the BAHD acyltransferase family, and PvCYP98A101 contained an ORF of 1530 bp encoding a 510-amino-acid residue, as a member of the CYP98A family. The deduced PvRAS and PvCYP98A101 amino acid sequences shared high similarity with other putative/known RASs and CYPs, respectively. Quantitative real-time PCR analysis showed that constitutive expression of PvRAS and PvCYP98A101 was much higher in roots than in leaves, stems, or spikes. Further analysis indicated that PvRAS was localised in the cytosol and nucleus, whilst PvCYP98A101 existed as a membrane protein in the endoplasmic reticulum. These results highlight the RA biosynthesis pathway in P. vulgaris and, provide useful information to engineer natural products.