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Identifier |
000441204 |
Title |
Βελτιστοποίηση των συνθηκών παραγωγής του μονοτερπενίου β-φελλανδρενίου από το γενετικά τροποποιημένο κυανοβακτήριο Synechocystis sp. PCC 6803 |
Alternative Title |
Optimal conditions for the production of the monoterpene β-phellandrene by the genetically modified cyanobacterium Synechocystis sp. PCC 6803 |
Author
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Χρυσουλάκη, Ελένη Ε.
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Thesis advisor
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Γανωτάκης, Δημήτριος
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Reviewer
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Παυλίδης, Ιωάννης
Τσιώτης, Γεώργιος
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Abstract |
The main objective of this study was the finding of optimal conditions in order
to increase the heterologous production of the monoterpene β-phellandrene by
Synechocystis sp. PCC 6803 transformants. This cyanobacterium is a model organism,
in which the “photosynthesis to fuel” approach has been successfully applied. Thus, it
is a suitable microorganism for biotechnological exploitation and production of a
large number of products, by using only sunlight, water and carbon dioxide (CO2) as
raw materials.
Previous laboratory studies have extensively studied the growth,
photosynthetic activity and production of β-phellandrene by mutant strains of the
cyanobacterium, under conditions of high salinity or even under conditions that used
only 100% seawater and the experiment lasted ten days with measurement values
taken every two days. Both their growth and production were successful. Therefore,
in the first chapter of this work, the wild type of cyanobacterium was studied under
various conditions of enriched seawater, which is abundant on the planet. The aim
was to find the most suitable growth condition of the strain in order to then study this
condition in the mutant strains and what is the effect on the production of the desired
product. The conditions that stood out from those studied were those with enriched
seawater with all the components of BG11 and HEPES / NaOH as buffer with pH 7.5
and enriched seawater with some components of BG11 with the same buffer. Under
these conditions the growth simulates to a very good extent the corresponding growth
of the WT strain in the control condition-BG11. The same results were seen in
photosynthetic activity. Therefore, in order to have a fairly economical method in the
rest of our study, as a condition to be applied to mutant strains was chosen, the one
with enriched seawater with some components present in the nutrient medium BG11.
In the next chapter, the mutants Δcpc+cpcB.PHLS+cpcA.GPPS and
Δcpc+cpcB.PHLS+cpc(-cpcA) were studied for growth, photosynthetic activity and
production of β-phellandrene in the optimal condition of seawater with some
components of BG11. The experiments lasted ten days and the measurements were
performed every second day. The results showed great similarity to the control
condition in terms of growth and photosynthetic activity. The yield of β-phellandrene
seemed to be lower than the control condition-BG11, but the cells are still able to
produce it successfully.
Finally, in the last chapter the constructs Δcpc+cpcB.PHLS+cpc(-cpcA) and
Δcpc+cpcB.PHLS+cpcA.GPPS+cpc were immobilized in alginate polymeric spheres
in the condition with enriched seawater for 12 days and β-phellandrene production
was measured, always compared to the production of the same construct in free cells
under the same condition. A previous study in the laboratory showed that
immobilization of the cells of the construct Δcpc+cpcB.PHLS+cpc(-cpcA) gives
higher production of β-phellandrene compared to the free cells in the nutrient medium
BG11, but also in 100% seawater. Immobilization of cells in enriched seawater
seems to have lower yields than free cells. These two strains studied remain
metabolically active for the period of twelve days of studied and the production of β--
phellandrene is possible.
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Language |
Greek |
Subject |
Biofuels |
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Cpc operon |
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Cpc οπερόνιο |
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Cyanobacteria |
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Fusion proteins |
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Geranyl diphosphate synthase (GPPS) |
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Metabolic engineering |
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Phellandrene synthase (PHLS) |
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Photosynthesis |
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Terpenoids |
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Βιοκαύσιμα |
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Κυανοβακτήρια |
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Μεταβολική μηχανική |
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Πρωτεΐνες σύντηξης |
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Συνθάση του πυροφωσφορικού γερανυλίου (GPPS) |
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Συνθάση του φελλανδρενίου (PHLS) |
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Τερπενοειδή |
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Φωτοσύνθεση |
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β-φελλανδρένιο |
Issue date |
2021-07-30 |
Collection
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School/Department--School of Sciences and Engineering--Department of Chemistry--Post-graduate theses
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Type of Work--Post-graduate theses
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Permanent Link |
https://elocus.lib.uoc.gr//dlib/d/1/4/metadata-dlib-1626174247-589188-3268.tkl
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Views |
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