Staff View: Metabolic alteration of Catharanthus roseus cell suspension cultures overexpressing geraniol synthase in the plastids or cytosol

Metabolic alteration of Catharanthus roseus cell suspension cultures overexpressing geraniol synthase in the plastids or cytosol

Previous studies showed that geraniol could be an upstream limiting factor in the monoterpenoid pathway towards the production of terpenoid indole alkaloid (TIA) in Catharanthus roseus cells and hairy root cultures. This shortage in precursor availability could be due to (1) limited expression of th...

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Main Authors:	Saiman, Mohd Zuwairi, Miettinen, Karel, Mustafa, Natali Rianika, Choi, Young Hae, Verpoorte, Robert, Schulte, Anna Elisabeth
Format:	Article
Published:	Springer 2018
Subjects:	Q Science (General) QH Natural history
Online Access:	http://eprints.um.edu.my/20811/ https://doi.org/10.1007/s11240-018-1398-5
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spelling	my.um.eprints.208112019-04-05T08:04:58Z http://eprints.um.edu.my/20811/ Metabolic alteration of Catharanthus roseus cell suspension cultures overexpressing geraniol synthase in the plastids or cytosol Saiman, Mohd Zuwairi Miettinen, Karel Mustafa, Natali Rianika Choi, Young Hae Verpoorte, Robert Schulte, Anna Elisabeth Q Science (General) QH Natural history Previous studies showed that geraniol could be an upstream limiting factor in the monoterpenoid pathway towards the production of terpenoid indole alkaloid (TIA) in Catharanthus roseus cells and hairy root cultures. This shortage in precursor availability could be due to (1) limited expression of the plastidial geraniol synthase resulted in a low activity of the enzyme to catalyze the conversion of geranyl diphosphate to geraniol; or (2) the limitation of geraniol transport from plastids to cytosol. Therefore, in this study, C. roseus’s geraniol synthase (CrGES) gene was overexpressed in either plastids or cytosol of a non-TIA producing C. roseus cell line. The expression of CrGES in the plastids or cytosol was confirmed and the constitutive transformation lines were successfully established. A targeted metabolite analysis using HPLC shows that the transformed cell lines did not produce TIA or iridoid precursors unless elicited with jasmonic acid, as their parent cell line. This indicates a requirement for expression of additional, inducible pathway genes to reach production of TIA in this cell line. Interestingly, further analysis using NMR-based metabolomics reveals that the overexpression of CrGES impacts primary metabolism differently if expressed in the plastids or cytosol. The levels of valine, leucine, and some metabolites derived from the shikimate pathway, i.e. phenylalanine and tyrosine were significantly higher in the plastidial- but lower in the cytosolic-CrGES overexpressing cell lines. This result shows that overexpression of CrGES in the plastids or cytosol caused alteration of primary metabolism that associated to the plant cell growth and development. A comprehensive omics analysis is necessary to reveal the full effect of metabolic engineering. Springer 2018 Article PeerReviewed Saiman, Mohd Zuwairi and Miettinen, Karel and Mustafa, Natali Rianika and Choi, Young Hae and Verpoorte, Robert and Schulte, Anna Elisabeth (2018) Metabolic alteration of Catharanthus roseus cell suspension cultures overexpressing geraniol synthase in the plastids or cytosol. Plant Cell, Tissue and Organ Culture, 134 (1). pp. 41-53. ISSN 0167-6857 https://doi.org/10.1007/s11240-018-1398-5 doi:10.1007/s11240-018-1398-5
institution	Universiti Malaya
building	UM Library
collection	Institutional Repository
continent	Asia
country	Malaysia
content_provider	Universiti Malaya
content_source	UM Research Repository
url_provider	http://eprints.um.edu.my/
topic	Q Science (General) QH Natural history
spellingShingle	Q Science (General) QH Natural history Saiman, Mohd Zuwairi Miettinen, Karel Mustafa, Natali Rianika Choi, Young Hae Verpoorte, Robert Schulte, Anna Elisabeth Metabolic alteration of Catharanthus roseus cell suspension cultures overexpressing geraniol synthase in the plastids or cytosol
description	Previous studies showed that geraniol could be an upstream limiting factor in the monoterpenoid pathway towards the production of terpenoid indole alkaloid (TIA) in Catharanthus roseus cells and hairy root cultures. This shortage in precursor availability could be due to (1) limited expression of the plastidial geraniol synthase resulted in a low activity of the enzyme to catalyze the conversion of geranyl diphosphate to geraniol; or (2) the limitation of geraniol transport from plastids to cytosol. Therefore, in this study, C. roseus’s geraniol synthase (CrGES) gene was overexpressed in either plastids or cytosol of a non-TIA producing C. roseus cell line. The expression of CrGES in the plastids or cytosol was confirmed and the constitutive transformation lines were successfully established. A targeted metabolite analysis using HPLC shows that the transformed cell lines did not produce TIA or iridoid precursors unless elicited with jasmonic acid, as their parent cell line. This indicates a requirement for expression of additional, inducible pathway genes to reach production of TIA in this cell line. Interestingly, further analysis using NMR-based metabolomics reveals that the overexpression of CrGES impacts primary metabolism differently if expressed in the plastids or cytosol. The levels of valine, leucine, and some metabolites derived from the shikimate pathway, i.e. phenylalanine and tyrosine were significantly higher in the plastidial- but lower in the cytosolic-CrGES overexpressing cell lines. This result shows that overexpression of CrGES in the plastids or cytosol caused alteration of primary metabolism that associated to the plant cell growth and development. A comprehensive omics analysis is necessary to reveal the full effect of metabolic engineering.
format	Article
author	Saiman, Mohd Zuwairi Miettinen, Karel Mustafa, Natali Rianika Choi, Young Hae Verpoorte, Robert Schulte, Anna Elisabeth
author_facet	Saiman, Mohd Zuwairi Miettinen, Karel Mustafa, Natali Rianika Choi, Young Hae Verpoorte, Robert Schulte, Anna Elisabeth
author_sort	Saiman, Mohd Zuwairi
title	Metabolic alteration of Catharanthus roseus cell suspension cultures overexpressing geraniol synthase in the plastids or cytosol
title_short	Metabolic alteration of Catharanthus roseus cell suspension cultures overexpressing geraniol synthase in the plastids or cytosol
title_full	Metabolic alteration of Catharanthus roseus cell suspension cultures overexpressing geraniol synthase in the plastids or cytosol
title_fullStr	Metabolic alteration of Catharanthus roseus cell suspension cultures overexpressing geraniol synthase in the plastids or cytosol
title_full_unstemmed	Metabolic alteration of Catharanthus roseus cell suspension cultures overexpressing geraniol synthase in the plastids or cytosol
title_sort	metabolic alteration of catharanthus roseus cell suspension cultures overexpressing geraniol synthase in the plastids or cytosol
publisher	Springer
publishDate	2018
url	http://eprints.um.edu.my/20811/ https://doi.org/10.1007/s11240-018-1398-5
_version_	1643691386925481984
score	13.211869

Metabolic alteration of Catharanthus roseus cell suspension cultures overexpressing geraniol synthase in the plastids or cytosol

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