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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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Main Authors: Yeap, Yee Hung, Koay, Teng Wei, Wong, Hann Ling, Lim, Boon Hoe
Format: Article
Language:English
Published: Penerbit Universiti Kebangsaan Malaysia 2018
Online Access:http://journalarticle.ukm.my/12497/1/04%20Yee%20Hung%20Yeap.pdf
http://journalarticle.ukm.my/12497/
http://www.ukm.my/jsm/english_journals/vol47num10_2018/contentsVol47num10_2018.htm
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spelling my-ukm.journal.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
institution Universiti Kebangsaan Malaysia
building Perpustakaan Tun Sri Lanang Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Kebangsaan Malaysia
content_source UKM Journal Article Repository
url_provider http://journalarticle.ukm.my/
language English
description 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/12497/1/04%20Yee%20Hung%20Yeap.pdf
http://journalarticle.ukm.my/12497/
http://www.ukm.my/jsm/english_journals/vol47num10_2018/contentsVol47num10_2018.htm
_version_ 1643738805814951936
score 13.211869

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