Dissection of Synechococcus rubisco large subunit sections involved in holoenzyme formation in Escherichia coli by combinatorial section swapping and sequence analyses
Engineering the CO2-fixing enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) to improve photosynthesis has long been sought. Rubisco large subunits (RbcL) are highly-conserved but because of certain undefined sequence differences, plant Rubisco research cannot fully utilise the robust...
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Penerbit Universiti Kebangsaan Malaysia
2018
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ukm-124972019-01-28T08:53:25Z http://journalarticle.ukm.my/12497/ Dissection of Synechococcus rubisco large subunit sections involved in holoenzyme formation in Escherichia coli by combinatorial section swapping and sequence analyses Yeap, Yee Hung Koay, Teng Wei Wong, Hann Ling Lim, Boon Hoe Engineering the CO2-fixing enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) to improve photosynthesis has long been sought. Rubisco large subunits (RbcL) are highly-conserved but because of certain undefined sequence differences, plant Rubisco research cannot fully utilise the robust heterologous Escherichia coli expression system and its GroEL folding machinery. Previously, a series of chimeric cyanobacteria Synechococcus elongatus Rubisco, incorporated with sequences from the green alga Chlamydomonas reinhardtii, were expressed in E. coli; differences in RbcL sections essential for holoenzyme formation were pinpointed. In this study, the remaining sections, presumably not crucial for holoenzyme formation and also the small subunit (RbcS), are substituted to further ascertain the possible destabilising effects of multiple section mutations. To that end, combinations of Synechococcus RbcL Sections 1 (residues 1-47), 2 (residues 48-97), 5 (residues 198-247) and 10 (residues 448-472), and RbcS, were swapped with collinear Chlamydomonas sections and expressed in E. coli. Interestingly, only the chimera with Sections 1 and 2 together produces holoenzyme and an interaction network of complementing amino acid changes is delineated by crystal structure analysis. Furthermore, sequence-based analysis also highlighted possible GroEL binding site differences between the two RbcLs. Penerbit Universiti Kebangsaan Malaysia 2018-10 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/12497/1/04%20Yee%20Hung%20Yeap.pdf Yeap, Yee Hung and Koay, Teng Wei and Wong, Hann Ling and Lim, Boon Hoe (2018) Dissection of Synechococcus rubisco large subunit sections involved in holoenzyme formation in Escherichia coli by combinatorial section swapping and sequence analyses. Sains Malaysiana, 47 (10). pp. 2269-2289. ISSN 0126-6039 http://www.ukm.my/jsm/english_journals/vol47num10_2018/contentsVol47num10_2018.htm |
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Engineering the CO2-fixing enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) to improve photosynthesis has long been sought. Rubisco large subunits (RbcL) are highly-conserved but because of certain undefined sequence differences, plant Rubisco research cannot fully utilise the robust heterologous Escherichia coli expression system and its GroEL folding machinery. Previously, a series of chimeric cyanobacteria Synechococcus elongatus Rubisco, incorporated with sequences from the green alga Chlamydomonas reinhardtii, were expressed in E. coli; differences in RbcL sections essential for holoenzyme formation were pinpointed. In this study, the remaining sections, presumably not crucial for holoenzyme formation and also the small subunit (RbcS), are substituted to further ascertain the possible destabilising effects of multiple section mutations. To that end, combinations of Synechococcus RbcL Sections 1 (residues 1-47), 2 (residues 48-97), 5 (residues 198-247) and 10 (residues 448-472), and RbcS, were swapped with collinear Chlamydomonas sections and expressed in E. coli. Interestingly, only the chimera with Sections 1 and 2 together produces holoenzyme and an interaction network of complementing amino acid changes is delineated by crystal structure analysis. Furthermore, sequence-based analysis also highlighted possible GroEL binding site differences between the two RbcLs. |
format |
Article |
author |
Yeap, Yee Hung Koay, Teng Wei Wong, Hann Ling Lim, Boon Hoe |
spellingShingle |
Yeap, Yee Hung Koay, Teng Wei Wong, Hann Ling Lim, Boon Hoe Dissection of Synechococcus rubisco large subunit sections involved in holoenzyme formation in Escherichia coli by combinatorial section swapping and sequence analyses |
author_facet |
Yeap, Yee Hung Koay, Teng Wei Wong, Hann Ling Lim, Boon Hoe |
author_sort |
Yeap, Yee Hung |
title |
Dissection of Synechococcus rubisco large subunit sections involved in holoenzyme formation in Escherichia coli by combinatorial section
swapping and sequence analyses |
title_short |
Dissection of Synechococcus rubisco large subunit sections involved in holoenzyme formation in Escherichia coli by combinatorial section
swapping and sequence analyses |
title_full |
Dissection of Synechococcus rubisco large subunit sections involved in holoenzyme formation in Escherichia coli by combinatorial section
swapping and sequence analyses |
title_fullStr |
Dissection of Synechococcus rubisco large subunit sections involved in holoenzyme formation in Escherichia coli by combinatorial section
swapping and sequence analyses |
title_full_unstemmed |
Dissection of Synechococcus rubisco large subunit sections involved in holoenzyme formation in Escherichia coli by combinatorial section
swapping and sequence analyses |
title_sort |
dissection of synechococcus rubisco large subunit sections involved in holoenzyme formation in escherichia coli by combinatorial section
swapping and sequence analyses |
publisher |
Penerbit Universiti Kebangsaan Malaysia |
publishDate |
2018 |
url |
http://journalarticle.ukm.my/12497/ http://journalarticle.ukm.my/12497/ http://journalarticle.ukm.my/12497/1/04%20Yee%20Hung%20Yeap.pdf |
first_indexed |
2023-09-18T20:02:44Z |
last_indexed |
2023-09-18T20:02:44Z |
_version_ |
1777406957305462784 |