Optimisation of high-value isoprenoid production in plants : Potential strategies and insights into carotenoid sequestration. / Nogueira, Marilise.

2013. 326 p.

Research output: ThesisDoctoral Thesis

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@phdthesis{520a6d6807164d2088a2923e4104a53f,
title = "Optimisation of high-value isoprenoid production in plants: Potential strategies and insights into carotenoid sequestration",
abstract = "Carotenoids and ketocarotenoids are isoprenoid molecules, which represent one of the most widespread classes of natural pigments, found in animals, plants andmicroorganisms. Moreover, they have valuable antioxidant properties. Their healthbenefits and colorant aspects have led to attempts to elevate their level in foodstuffs.In the present study, several metabolic engineering strategies were tested in order to enhance the levels of high-value carotenoid and ketocarotenoid compounds, such as lycopene, -carotene, canthaxanthin and astaxanthin, in tomato and tobacco plants.Biosynthetic bacterial pathway genes have been overexpressed independently(GGPP synthase (CrtE), phytoene synthase (CrtB) and phytoene desaturase (CrtI))and in combination (CrtE+B, CrtE+I, CrtB+I and -carotene hydroxylase andketolase (CrtZ+W)), with different promoters (for CrtB, CrtI and CrtB+I) or inassociation with transcription factors (Phytochrome-interacting factor 5(CrtZ+W+PIF5) and Arabidopsis Response Regulator 14 (CrtZ+W+ARR14)). Theeffects of these different strategies on the plant metabolism and especially oncarotenoid formation, sequestration and the activation of regulation mechanismswere studied.The combination of the two genes CrtB and CrtI, in their hemizygous state, had asynergistic effect on the production of carotenoids and the expression ofCrtZ+W+ARR14 increased the levels of ketocarotenoids in the plants. The importantfeatures for the design of metabolic engineering strategies were highlighted.Moreover, regulatory mechanisms that operate across multiple levels of cellularregulation, including transcription, protein localisation, metabolite levels, cell ortissue type, and organelle/sub-organelle structure and organisation were revealed. It was demonstrated how changes to chromoplast and sub-chromoplast structures, such as crystal formation, plastoglobule and membrane composition/structures can arise in response to changes in metabolites. A new carotenoid regulation mechanism at the sub-organellar level was discovered and a schematic model was proposed.",
keywords = "carotenoid, ketocarotenoid, sequestration",
author = "Marilise Nogueira",
year = "2013",
doi = "10.1105/tpc.113.116210",
language = "English",
school = "Royal Holloway, University of London",

}

RIS

TY - THES

T1 - Optimisation of high-value isoprenoid production in plants

T2 - Potential strategies and insights into carotenoid sequestration

AU - Nogueira, Marilise

PY - 2013

Y1 - 2013

N2 - Carotenoids and ketocarotenoids are isoprenoid molecules, which represent one of the most widespread classes of natural pigments, found in animals, plants andmicroorganisms. Moreover, they have valuable antioxidant properties. Their healthbenefits and colorant aspects have led to attempts to elevate their level in foodstuffs.In the present study, several metabolic engineering strategies were tested in order to enhance the levels of high-value carotenoid and ketocarotenoid compounds, such as lycopene, -carotene, canthaxanthin and astaxanthin, in tomato and tobacco plants.Biosynthetic bacterial pathway genes have been overexpressed independently(GGPP synthase (CrtE), phytoene synthase (CrtB) and phytoene desaturase (CrtI))and in combination (CrtE+B, CrtE+I, CrtB+I and -carotene hydroxylase andketolase (CrtZ+W)), with different promoters (for CrtB, CrtI and CrtB+I) or inassociation with transcription factors (Phytochrome-interacting factor 5(CrtZ+W+PIF5) and Arabidopsis Response Regulator 14 (CrtZ+W+ARR14)). Theeffects of these different strategies on the plant metabolism and especially oncarotenoid formation, sequestration and the activation of regulation mechanismswere studied.The combination of the two genes CrtB and CrtI, in their hemizygous state, had asynergistic effect on the production of carotenoids and the expression ofCrtZ+W+ARR14 increased the levels of ketocarotenoids in the plants. The importantfeatures for the design of metabolic engineering strategies were highlighted.Moreover, regulatory mechanisms that operate across multiple levels of cellularregulation, including transcription, protein localisation, metabolite levels, cell ortissue type, and organelle/sub-organelle structure and organisation were revealed. It was demonstrated how changes to chromoplast and sub-chromoplast structures, such as crystal formation, plastoglobule and membrane composition/structures can arise in response to changes in metabolites. A new carotenoid regulation mechanism at the sub-organellar level was discovered and a schematic model was proposed.

AB - Carotenoids and ketocarotenoids are isoprenoid molecules, which represent one of the most widespread classes of natural pigments, found in animals, plants andmicroorganisms. Moreover, they have valuable antioxidant properties. Their healthbenefits and colorant aspects have led to attempts to elevate their level in foodstuffs.In the present study, several metabolic engineering strategies were tested in order to enhance the levels of high-value carotenoid and ketocarotenoid compounds, such as lycopene, -carotene, canthaxanthin and astaxanthin, in tomato and tobacco plants.Biosynthetic bacterial pathway genes have been overexpressed independently(GGPP synthase (CrtE), phytoene synthase (CrtB) and phytoene desaturase (CrtI))and in combination (CrtE+B, CrtE+I, CrtB+I and -carotene hydroxylase andketolase (CrtZ+W)), with different promoters (for CrtB, CrtI and CrtB+I) or inassociation with transcription factors (Phytochrome-interacting factor 5(CrtZ+W+PIF5) and Arabidopsis Response Regulator 14 (CrtZ+W+ARR14)). Theeffects of these different strategies on the plant metabolism and especially oncarotenoid formation, sequestration and the activation of regulation mechanismswere studied.The combination of the two genes CrtB and CrtI, in their hemizygous state, had asynergistic effect on the production of carotenoids and the expression ofCrtZ+W+ARR14 increased the levels of ketocarotenoids in the plants. The importantfeatures for the design of metabolic engineering strategies were highlighted.Moreover, regulatory mechanisms that operate across multiple levels of cellularregulation, including transcription, protein localisation, metabolite levels, cell ortissue type, and organelle/sub-organelle structure and organisation were revealed. It was demonstrated how changes to chromoplast and sub-chromoplast structures, such as crystal formation, plastoglobule and membrane composition/structures can arise in response to changes in metabolites. A new carotenoid regulation mechanism at the sub-organellar level was discovered and a schematic model was proposed.

KW - carotenoid

KW - ketocarotenoid

KW - sequestration

U2 - 10.1105/tpc.113.116210

DO - 10.1105/tpc.113.116210

M3 - Doctoral Thesis

ER -