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Determination of the role of synechococcus rubisco residue val-425 in chimeras

Ribulose-1,5-bisphosphate carboxylase/oxygenase (EC 4.1.1.39), Rubisco is one of the most extensively studied enzymes due to its abundance on Earth and its significance in the food supply chain. As the rate-limiting enzyme in the Calvin cycle, the catalytic inefficiency of Rubisco has led to an unde...

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Main Author: Wong, Clara Chia Ci
Format: Final Year Project / Dissertation / Thesis
Published: 2023
Subjects:
Q Science (General)
QD Chemistry
QR Microbiology
Online Access:http://eprints.utar.edu.my/5924/1/2006159_Thesis_%2D_WONG_CHIA_CI_CLARA.pdf
http://eprints.utar.edu.my/5924/
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spelling my-utar-eprints.59242024-01-03T10:46:20Z Determination of the role of synechococcus rubisco residue val-425 in chimeras Wong, Clara Chia Ci Q Science (General) QD Chemistry QR Microbiology Ribulose-1,5-bisphosphate carboxylase/oxygenase (EC 4.1.1.39), Rubisco is one of the most extensively studied enzymes due to its abundance on Earth and its significance in the food supply chain. As the rate-limiting enzyme in the Calvin cycle, the catalytic inefficiency of Rubisco has led to an undesirable side reaction known as photorespiration that compromises the carboxylase activity of CO2 fixation, thus, making it the primary target for bioengineering to improve its catalytic efficiency. Innovative approaches were made by researchers to improve the carboxylation capacity of Rubisco which nonetheless was hampered by the unsuccessful heterologous formation of functional Rubisco enzyme, plausibly due to incompatible assembly pathways. Previous findings by Koay, Wong and Lim (2016) were extended in this study whereby the chimeric mutant bearing mutated residue 425 (Thr-425) is used as the target for site-directed mutagenesis (SDM) that reverts the Thr mutation to Val residue. The current study explores the potential role of Val-425 in the large subunit (RbcL) of Synechococcus Rubisco that is previously hypothesized to be critical for the chaperone-mediated assembly of the holoenzyme. SDM had successfully restored Val residue at position 425 of RbcL which was validated by restriction iii digestion of PstI enzyme. Expression for both Rubisco subunits (RbcL and RbcS) upon SDM was detected on SDS-PAGE at 55 kDa and below 15 kDa, respectively. Furthermore, Val-425 is also found to be important in conferring stability in the native conformation of Synechococcus RbcL and its contribution to greater hydrophobicity suggests its associated role in chaperonin-binding. 2023-06 Final Year Project / Dissertation / Thesis NonPeerReviewed application/pdf http://eprints.utar.edu.my/5924/1/2006159_Thesis_%2D_WONG_CHIA_CI_CLARA.pdf Wong, Clara Chia Ci (2023) Determination of the role of synechococcus rubisco residue val-425 in chimeras. Final Year Project, UTAR. http://eprints.utar.edu.my/5924/
institution Universiti Tunku Abdul Rahman
building UTAR Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Tunku Abdul Rahman
content_source UTAR Institutional Repository
url_provider http://eprints.utar.edu.my
topic Q Science (General)
QD Chemistry
QR Microbiology
spellingShingle Q Science (General)
QD Chemistry
QR Microbiology
Wong, Clara Chia Ci
Determination of the role of synechococcus rubisco residue val-425 in chimeras
description Ribulose-1,5-bisphosphate carboxylase/oxygenase (EC 4.1.1.39), Rubisco is one of the most extensively studied enzymes due to its abundance on Earth and its significance in the food supply chain. As the rate-limiting enzyme in the Calvin cycle, the catalytic inefficiency of Rubisco has led to an undesirable side reaction known as photorespiration that compromises the carboxylase activity of CO2 fixation, thus, making it the primary target for bioengineering to improve its catalytic efficiency. Innovative approaches were made by researchers to improve the carboxylation capacity of Rubisco which nonetheless was hampered by the unsuccessful heterologous formation of functional Rubisco enzyme, plausibly due to incompatible assembly pathways. Previous findings by Koay, Wong and Lim (2016) were extended in this study whereby the chimeric mutant bearing mutated residue 425 (Thr-425) is used as the target for site-directed mutagenesis (SDM) that reverts the Thr mutation to Val residue. The current study explores the potential role of Val-425 in the large subunit (RbcL) of Synechococcus Rubisco that is previously hypothesized to be critical for the chaperone-mediated assembly of the holoenzyme. SDM had successfully restored Val residue at position 425 of RbcL which was validated by restriction iii digestion of PstI enzyme. Expression for both Rubisco subunits (RbcL and RbcS) upon SDM was detected on SDS-PAGE at 55 kDa and below 15 kDa, respectively. Furthermore, Val-425 is also found to be important in conferring stability in the native conformation of Synechococcus RbcL and its contribution to greater hydrophobicity suggests its associated role in chaperonin-binding.
format Final Year Project / Dissertation / Thesis
author Wong, Clara Chia Ci
author_facet Wong, Clara Chia Ci
author_sort Wong, Clara Chia Ci
title Determination of the role of synechococcus rubisco residue val-425 in chimeras
title_short Determination of the role of synechococcus rubisco residue val-425 in chimeras
title_full Determination of the role of synechococcus rubisco residue val-425 in chimeras
title_fullStr Determination of the role of synechococcus rubisco residue val-425 in chimeras
title_full_unstemmed Determination of the role of synechococcus rubisco residue val-425 in chimeras
title_sort determination of the role of synechococcus rubisco residue val-425 in chimeras
publishDate 2023
url http://eprints.utar.edu.my/5924/1/2006159_Thesis_%2D_WONG_CHIA_CI_CLARA.pdf
http://eprints.utar.edu.my/5924/
_version_ 1787519964528246784
score 13.211869

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