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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 18, No. 5, 2015, pp. 376-380
Bioline Code: ej15062
Full paper language: English
Document type: Research Article
Document available free of charge

Electronic Journal of Biotechnology, Vol. 18, No. 5, 2015, pp. 376-380

 en Isolation and characterization of a novel (S)-canadine synthase gene from Coptis chinensis check for this species in other resources
He, Yang; Hou, Pei; Fan, Gang; Li, Dongmei & Peng, Cheng


Background: Berberine acts primarily as an important active ingredient in Coptis chinensis check for this species in other resources and has been traditionally applied in clinical treatment. Nevertheless, little information has been released about C. chinensis, as far as functional genes and biosynthetic pathway of berberine are concerned. Here, we isolated a novel (S)-canadine synthase gene (designated as CAS-1) from C. chinensis by using RT-PCR and RACE techniques.
Results: Bioinformatics analysis showed that the cDNA is 1942 bp in length with a complete open reading frame (ORF) of 1476 bp, and the ORF encodes a polypeptide of 491 amino acids with a calculated molecular mass of 55.29 kDa and a pI value of 8.92. Real-time quantitative PCR analysis showed that CcCAS-1 was constitutively expressed in leaf, petiole and rhizome tissues, especially in the leaves of C. chinensis. However, the results of berberine content in different tissues by high-performance liquid chromatography with photodiode array detection (HPLC-DAD)method showed that the leaves and the petiole tissues have similar content of berberine.
Conclusions: We found that the berberine content in the rhizomewas seven times (more or less) than that in the leaves and the petioles. In addition, the full length coding sequence of CcCAS-1 was inserted into pET-32a andwas successfully expressed in Escherichia coli check for this species in other resources , laying a solid foundation for protein purification, activity assay and multi-clonal antibody preparation. Together, our data suggest that CcCAS-1 is a novel heme-thiolate enzyme essential for berberine biosynthesis in C. chinensis.

Berberine biosynthesis; Coptis chinensis; Expression pattern

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