Abstract
Macleaya cordata produces a variety of benzylisoquinoline alkaloids (BIAs), such as sanguinarine, protopine, and berberine, which are potential anticancer drugs and natural growth promoters. The genes encoding the berberine bridge enzyme (BBE) were isolated from M. cordata and Papaver somniferum, and then the two genes were overexpressed in M. cordata. Through liquid chromatography with triple-quadrupole mass spectrometry analysis, it was determined that McBBE-OX caused higher levels of (S)-norcoclaurine, (S)-coclaurine, (S)-N-cis-methylcoclaurine, (S)-reticuline, (S)-tetrahydrocolumbamine, (S)-tetrahydroberberine, (S)-cheilanthifoline, and (S)-scoulerine than PsBBE-OX, empty vector or control treatments. qRT-PCR analysis demonstrated that the introduced genes in the transgenic lines were all highly expressed. However, the levels of sanguinarine (SAN) and chelerythrine (CHE) in all the transgenic lines were slightly lower than those in the wild-type lines, possibly because the overexpression of McBBE causes feedback-inhibition. This is the first report on the overexpression of potential key genes in M. cordata, and the findings are important for the design of metabolic engineering strategies that target BIAs biosynthesis.
Highlights
Benzylisoquinoline alkaloids (BIAs) are a large and structurally diverse group of natural products, most of which occur in the families Papaveraceae, Ranunculaceae, Lauraceae, Rutaceae and Menispermaceae[1]
In a previous study, the content of benzylisoquinoline alkaloids (BIAs) in California poppy was increased through overexpression of the P. somniferum berberine bridge enzyme (BBE) gene (PsBBE), and this experiment indicated that BBE is a potential key enzyme to enhance the content of BIAs12
We investigated whether the overexpression of McBBE or PsBBE in M. cordata could affect the biosynthesis of BIAs
Summary
Benzylisoquinoline alkaloids (BIAs) are a large and structurally diverse group of natural products, most of which occur in the families Papaveraceae, Ranunculaceae, Lauraceae, Rutaceae and Menispermaceae[1]. This work solved the problem of BIA production needing precursors, but the synthesis step was too long, and the final product concentration was hardly sufficient for industrial production For this reason, many studies have used transgenic methods to increase production, especially overexpression of potential key enzymes. We investigated whether the overexpression of McBBE or PsBBE in M. cordata could affect the biosynthesis of BIAs. we investigated the metabolite profiles through ESI/QQQ MS analysis and detected the expression levels of 11 genes involved in the synthesis of SAN and CHE. We investigated the metabolite profiles through ESI/QQQ MS analysis and detected the expression levels of 11 genes involved in the synthesis of SAN and CHE To our knowledge, this is the first study using overexpression of BBE in transgenic M. cordata with increased alkaloid production
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