Metabolic engineering and elicitation of pharmaceutically active metabolites in Rhazya stricta (Apocynaceau)

Amir Akhgari, I. Laakso, T. Yrjönen, H. Vuorela, Kirsi-Marja Oksman-Caldentey, Heiko Rischer

Research output: Contribution to journalOther journal contributionScientific

Abstract

Rhazya stricta Decne., a small evergreen shrub native to the Middle East and Indian sub-continent, is a rich source of pharmacologically active terpenoid indole alkaloids (TIAs). The plant has been in the focus of phytochemical, pharmacological and ethnobotanical studies due to its long use in folk medicine to treat a variety of ailments. More recently especially the antimicrobial and anticancer properties of certain constituents (Fig. 1) have been established. TIAs are formed by the condensation of the indole and iridoid moieties catalysed by strictosidine synthase (STR) to form the precursor strictosidine. High STR activity associated with the induction of rate-limiting enzyme activities of the iridoid pathway (GES, G10 H) favours TIA production. Biosynthetic steps in the TIA pathway are co-ordinately regulated at the level of transcription of the structural genes by transcription factors (e.g. orca3). Wild type hairy root cultures were established in order to study TIA production and enable metabolic engineering. A validated GC-MS method was developed and 20 compounds were identified. Rhazinilam, rhazine and vallesiachotamine were present in several lines (Fig. 1). Gene(s) from the early part of the TIA pathway, single gene geraniol synthase (ges), geraniol 10-hydroxylase (g10h) and strictosidine synthase (str) and double-gene (str+orca3), were introduced to R. stricta to investigate their over-expression effects on the TIA biosynthesis in transgenic hairy root lines. Integration and expression of the gene(s) were confirmed by molecular techniques. Suitable elicitations by methyl jasmonate to stimulate TIA accumulation were evaluated. Chemical analysis of transformed hairy roots and elicited samples showed an increased level of selected TIAs compared to the wild type hairy root lines and non-elicited samples. Further analysis revealed higher accumulation of total alkaloids in transformed and elicited hairy roots.
Original languageEnglish
Article numberPM1
JournalPlanta Medica
Volume79
Issue number13
DOIs
Publication statusPublished - 2013
MoE publication typeNot Eligible
Event61st International Congress and Annual Meeting of the Society for Medicinal Plant and Natural Product Research - Münster, Germany
Duration: 1 Sep 20135 Sep 2013
Conference number: 61

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Secologanin Tryptamine Alkaloids
Apocynaceae
Metabolic engineering
Metabolic Engineering
Metabolites
Genes
Middle East
Biosynthesis
Enzyme activity
Phytochemicals
Traditional Medicine
Transcription
Alkaloids
Medicine
Condensation
Transcription Factors
Pharmacology

Cite this

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title = "Metabolic engineering and elicitation of pharmaceutically active metabolites in Rhazya stricta (Apocynaceau)",
abstract = "Rhazya stricta Decne., a small evergreen shrub native to the Middle East and Indian sub-continent, is a rich source of pharmacologically active terpenoid indole alkaloids (TIAs). The plant has been in the focus of phytochemical, pharmacological and ethnobotanical studies due to its long use in folk medicine to treat a variety of ailments. More recently especially the antimicrobial and anticancer properties of certain constituents (Fig. 1) have been established. TIAs are formed by the condensation of the indole and iridoid moieties catalysed by strictosidine synthase (STR) to form the precursor strictosidine. High STR activity associated with the induction of rate-limiting enzyme activities of the iridoid pathway (GES, G10 H) favours TIA production. Biosynthetic steps in the TIA pathway are co-ordinately regulated at the level of transcription of the structural genes by transcription factors (e.g. orca3). Wild type hairy root cultures were established in order to study TIA production and enable metabolic engineering. A validated GC-MS method was developed and 20 compounds were identified. Rhazinilam, rhazine and vallesiachotamine were present in several lines (Fig. 1). Gene(s) from the early part of the TIA pathway, single gene geraniol synthase (ges), geraniol 10-hydroxylase (g10h) and strictosidine synthase (str) and double-gene (str+orca3), were introduced to R. stricta to investigate their over-expression effects on the TIA biosynthesis in transgenic hairy root lines. Integration and expression of the gene(s) were confirmed by molecular techniques. Suitable elicitations by methyl jasmonate to stimulate TIA accumulation were evaluated. Chemical analysis of transformed hairy roots and elicited samples showed an increased level of selected TIAs compared to the wild type hairy root lines and non-elicited samples. Further analysis revealed higher accumulation of total alkaloids in transformed and elicited hairy roots.",
author = "Amir Akhgari and I. Laakso and T. Yrj{\"o}nen and H. Vuorela and Kirsi-Marja Oksman-Caldentey and Heiko Rischer",
note = "Only abstract published",
year = "2013",
doi = "10.1055/s-0033-1352334",
language = "English",
volume = "79",
journal = "Planta Medica",
issn = "0032-0943",
publisher = "Georg Thieme Verlag",
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Metabolic engineering and elicitation of pharmaceutically active metabolites in Rhazya stricta (Apocynaceau). / Akhgari, Amir; Laakso, I.; Yrjönen, T.; Vuorela, H.; Oksman-Caldentey, Kirsi-Marja; Rischer, Heiko.

In: Planta Medica, Vol. 79, No. 13, PM1, 2013.

Research output: Contribution to journalOther journal contributionScientific

TY - JOUR

T1 - Metabolic engineering and elicitation of pharmaceutically active metabolites in Rhazya stricta (Apocynaceau)

AU - Akhgari, Amir

AU - Laakso, I.

AU - Yrjönen, T.

AU - Vuorela, H.

AU - Oksman-Caldentey, Kirsi-Marja

AU - Rischer, Heiko

N1 - Only abstract published

PY - 2013

Y1 - 2013

N2 - Rhazya stricta Decne., a small evergreen shrub native to the Middle East and Indian sub-continent, is a rich source of pharmacologically active terpenoid indole alkaloids (TIAs). The plant has been in the focus of phytochemical, pharmacological and ethnobotanical studies due to its long use in folk medicine to treat a variety of ailments. More recently especially the antimicrobial and anticancer properties of certain constituents (Fig. 1) have been established. TIAs are formed by the condensation of the indole and iridoid moieties catalysed by strictosidine synthase (STR) to form the precursor strictosidine. High STR activity associated with the induction of rate-limiting enzyme activities of the iridoid pathway (GES, G10 H) favours TIA production. Biosynthetic steps in the TIA pathway are co-ordinately regulated at the level of transcription of the structural genes by transcription factors (e.g. orca3). Wild type hairy root cultures were established in order to study TIA production and enable metabolic engineering. A validated GC-MS method was developed and 20 compounds were identified. Rhazinilam, rhazine and vallesiachotamine were present in several lines (Fig. 1). Gene(s) from the early part of the TIA pathway, single gene geraniol synthase (ges), geraniol 10-hydroxylase (g10h) and strictosidine synthase (str) and double-gene (str+orca3), were introduced to R. stricta to investigate their over-expression effects on the TIA biosynthesis in transgenic hairy root lines. Integration and expression of the gene(s) were confirmed by molecular techniques. Suitable elicitations by methyl jasmonate to stimulate TIA accumulation were evaluated. Chemical analysis of transformed hairy roots and elicited samples showed an increased level of selected TIAs compared to the wild type hairy root lines and non-elicited samples. Further analysis revealed higher accumulation of total alkaloids in transformed and elicited hairy roots.

AB - Rhazya stricta Decne., a small evergreen shrub native to the Middle East and Indian sub-continent, is a rich source of pharmacologically active terpenoid indole alkaloids (TIAs). The plant has been in the focus of phytochemical, pharmacological and ethnobotanical studies due to its long use in folk medicine to treat a variety of ailments. More recently especially the antimicrobial and anticancer properties of certain constituents (Fig. 1) have been established. TIAs are formed by the condensation of the indole and iridoid moieties catalysed by strictosidine synthase (STR) to form the precursor strictosidine. High STR activity associated with the induction of rate-limiting enzyme activities of the iridoid pathway (GES, G10 H) favours TIA production. Biosynthetic steps in the TIA pathway are co-ordinately regulated at the level of transcription of the structural genes by transcription factors (e.g. orca3). Wild type hairy root cultures were established in order to study TIA production and enable metabolic engineering. A validated GC-MS method was developed and 20 compounds were identified. Rhazinilam, rhazine and vallesiachotamine were present in several lines (Fig. 1). Gene(s) from the early part of the TIA pathway, single gene geraniol synthase (ges), geraniol 10-hydroxylase (g10h) and strictosidine synthase (str) and double-gene (str+orca3), were introduced to R. stricta to investigate their over-expression effects on the TIA biosynthesis in transgenic hairy root lines. Integration and expression of the gene(s) were confirmed by molecular techniques. Suitable elicitations by methyl jasmonate to stimulate TIA accumulation were evaluated. Chemical analysis of transformed hairy roots and elicited samples showed an increased level of selected TIAs compared to the wild type hairy root lines and non-elicited samples. Further analysis revealed higher accumulation of total alkaloids in transformed and elicited hairy roots.

U2 - 10.1055/s-0033-1352334

DO - 10.1055/s-0033-1352334

M3 - Other journal contribution

VL - 79

JO - Planta Medica

JF - Planta Medica

SN - 0032-0943

IS - 13

M1 - PM1

ER -