The final aim of this thesis was to achieve the encapsulation of photosynthetic reaction centers
with light harvesting complex 1 from Rhodobacter sphaeroides into viral capsids of
bacteriophage P22. To this end, the reaction centers and the capsids had to be isolated and
purified first. The capsids are composed of a coat protein forming the stable outer shell and a
scaffolding protein driving the self-assembly of the capsids and forming its inner lining.
Therefore, scaffolding proteins could be used to introduce the reaction center into the capsids.
To achieve that, the scaffolding protein was successfully functionalized with an NTA-
maleimide linker for binding with the HisTag of the reaction center's H subunit, creating a
reaction center-scaffolding protein complex. By inducing self-assembly of the capsid's
components with this complex, the reaction centers were successfully encapsulated. This was
confirmed by transmission electron microscopy pictures showing the encapsulated reaction
centers. These engineered virus-like particles containing photosystems are therefore a first
step towards the assembly of a nanoreactor featuring a photosynthetic reaction center
powering a cascade of chemical reactions performed by additionally co-encapsulated
enzymes.
Anotace v angličtině
The final aim of this thesis was to achieve the encapsulation of photosynthetic reaction centers
with light harvesting complex 1 from Rhodobacter sphaeroides into viral capsids of
bacteriophage P22. To this end, the reaction centers and the capsids had to be isolated and
purified first. The capsids are composed of a coat protein forming the stable outer shell and a
scaffolding protein driving the self-assembly of the capsids and forming its inner lining.
Therefore, scaffolding proteins could be used to introduce the reaction center into the capsids.
To achieve that, the scaffolding protein was successfully functionalized with an NTA-
maleimide linker for binding with the HisTag of the reaction center's H subunit, creating a
reaction center-scaffolding protein complex. By inducing self-assembly of the capsid's
components with this complex, the reaction centers were successfully encapsulated. This was
confirmed by transmission electron microscopy pictures showing the encapsulated reaction
centers. These engineered virus-like particles containing photosystems are therefore a first
step towards the assembly of a nanoreactor featuring a photosynthetic reaction center
powering a cascade of chemical reactions performed by additionally co-encapsulated
enzymes.
Klíčová slova
photosynthesis, reaction center, Rhodobacter sphaeroides, viral capsids, bacteriophage P22, nanobioreactor, scaffolding protein, coat protein, NTA maleimide, thrombin
Klíčová slova v angličtině
photosynthesis, reaction center, Rhodobacter sphaeroides, viral capsids, bacteriophage P22, nanobioreactor, scaffolding protein, coat protein, NTA maleimide, thrombin
Rozsah průvodní práce
37
Jazyk
AN
Anotace
The final aim of this thesis was to achieve the encapsulation of photosynthetic reaction centers
with light harvesting complex 1 from Rhodobacter sphaeroides into viral capsids of
bacteriophage P22. To this end, the reaction centers and the capsids had to be isolated and
purified first. The capsids are composed of a coat protein forming the stable outer shell and a
scaffolding protein driving the self-assembly of the capsids and forming its inner lining.
Therefore, scaffolding proteins could be used to introduce the reaction center into the capsids.
To achieve that, the scaffolding protein was successfully functionalized with an NTA-
maleimide linker for binding with the HisTag of the reaction center's H subunit, creating a
reaction center-scaffolding protein complex. By inducing self-assembly of the capsid's
components with this complex, the reaction centers were successfully encapsulated. This was
confirmed by transmission electron microscopy pictures showing the encapsulated reaction
centers. These engineered virus-like particles containing photosystems are therefore a first
step towards the assembly of a nanoreactor featuring a photosynthetic reaction center
powering a cascade of chemical reactions performed by additionally co-encapsulated
enzymes.
Anotace v angličtině
The final aim of this thesis was to achieve the encapsulation of photosynthetic reaction centers
with light harvesting complex 1 from Rhodobacter sphaeroides into viral capsids of
bacteriophage P22. To this end, the reaction centers and the capsids had to be isolated and
purified first. The capsids are composed of a coat protein forming the stable outer shell and a
scaffolding protein driving the self-assembly of the capsids and forming its inner lining.
Therefore, scaffolding proteins could be used to introduce the reaction center into the capsids.
To achieve that, the scaffolding protein was successfully functionalized with an NTA-
maleimide linker for binding with the HisTag of the reaction center's H subunit, creating a
reaction center-scaffolding protein complex. By inducing self-assembly of the capsid's
components with this complex, the reaction centers were successfully encapsulated. This was
confirmed by transmission electron microscopy pictures showing the encapsulated reaction
centers. These engineered virus-like particles containing photosystems are therefore a first
step towards the assembly of a nanoreactor featuring a photosynthetic reaction center
powering a cascade of chemical reactions performed by additionally co-encapsulated
enzymes.
Klíčová slova
photosynthesis, reaction center, Rhodobacter sphaeroides, viral capsids, bacteriophage P22, nanobioreactor, scaffolding protein, coat protein, NTA maleimide, thrombin
Klíčová slova v angličtině
photosynthesis, reaction center, Rhodobacter sphaeroides, viral capsids, bacteriophage P22, nanobioreactor, scaffolding protein, coat protein, NTA maleimide, thrombin
Zásady pro vypracování
-
Zásady pro vypracování
-
Seznam doporučené literatury
-
Seznam doporučené literatury
-
Přílohy volně vložené
-
Přílohy vázané v práci
ilustrace, grafy, schémata, tabulky
Převzato z knihovny
Ne
Plný text práce
Přílohy
Posudek(y) oponenta
Hodnocení vedoucího
Záznam průběhu obhajoby
Prof Grubhoffer welcomed the student and commission members; opponent Dr Malinovská was not present. The student presented the theoretical background of his work on the studied phage and the encapsulation, followed by the experimental part and the results. The supervisor, Dr Kaftan, presented his review, and the opponent review was read by Dr Štěrba. The student answered all the questions of the opponent and also questions from the commission members on the encapsulation process, expression of proteins, yield of production, future steps of the thesis.