Recombining low homology, functionally rich regions of bacterial subtilisins by combinatorial fragment exchange.
Combinatorial fragment exchange was utilised to recombine key structural and functional low homology regions of bacilli subtilisins to generate new active hybrid proteases with altered substrate profiles. Up to six different regions comprising mostly of loop residues from the commercially important...
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Public Library of Science (PLoS)
2011-01-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0024319&type=printable |
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| author | D Dafydd Jones |
| author_facet | D Dafydd Jones |
| author_sort | D Dafydd Jones |
| collection | DOAJ |
| description | Combinatorial fragment exchange was utilised to recombine key structural and functional low homology regions of bacilli subtilisins to generate new active hybrid proteases with altered substrate profiles. Up to six different regions comprising mostly of loop residues from the commercially important subtilisin Savinase were exchanged with the structurally equivalent regions of six other subtilisins. The six additional subtilisins derive from diverse origins and included thermophilic and intracellular subtilisins as well as other academically and commercially relevant subtilisins. Savinase was largely tolerant to fragment exchange; rational replacement of all six regions with 5 of 6 donating subtilisin sequences preserved activity, albeit reduced compared to Savinase. A combinatorial approach was used to generate hybrid Savinase variants in which the sequences derived from all seven subtilisins at each region were recombined to generate new region combinations. Variants with different substrate profiles and with greater apparent activity compared to Savinase and the rational fragment exchange variants were generated with the substrate profile exhibited by variants dependent on the sequence combination at each region. |
| format | Article |
| id | doaj-art-e7616f689c3e4f718af7ebea36fc00b3 |
| institution | Kabale University |
| issn | 1932-6203 |
| language | English |
| publishDate | 2011-01-01 |
| publisher | Public Library of Science (PLoS) |
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| series | PLoS ONE |
| spelling | doaj-art-e7616f689c3e4f718af7ebea36fc00b32025-08-20T03:25:08ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-0169e2431910.1371/journal.pone.0024319Recombining low homology, functionally rich regions of bacterial subtilisins by combinatorial fragment exchange.D Dafydd JonesCombinatorial fragment exchange was utilised to recombine key structural and functional low homology regions of bacilli subtilisins to generate new active hybrid proteases with altered substrate profiles. Up to six different regions comprising mostly of loop residues from the commercially important subtilisin Savinase were exchanged with the structurally equivalent regions of six other subtilisins. The six additional subtilisins derive from diverse origins and included thermophilic and intracellular subtilisins as well as other academically and commercially relevant subtilisins. Savinase was largely tolerant to fragment exchange; rational replacement of all six regions with 5 of 6 donating subtilisin sequences preserved activity, albeit reduced compared to Savinase. A combinatorial approach was used to generate hybrid Savinase variants in which the sequences derived from all seven subtilisins at each region were recombined to generate new region combinations. Variants with different substrate profiles and with greater apparent activity compared to Savinase and the rational fragment exchange variants were generated with the substrate profile exhibited by variants dependent on the sequence combination at each region.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0024319&type=printable |
| spellingShingle | D Dafydd Jones Recombining low homology, functionally rich regions of bacterial subtilisins by combinatorial fragment exchange. PLoS ONE |
| title | Recombining low homology, functionally rich regions of bacterial subtilisins by combinatorial fragment exchange. |
| title_full | Recombining low homology, functionally rich regions of bacterial subtilisins by combinatorial fragment exchange. |
| title_fullStr | Recombining low homology, functionally rich regions of bacterial subtilisins by combinatorial fragment exchange. |
| title_full_unstemmed | Recombining low homology, functionally rich regions of bacterial subtilisins by combinatorial fragment exchange. |
| title_short | Recombining low homology, functionally rich regions of bacterial subtilisins by combinatorial fragment exchange. |
| title_sort | recombining low homology functionally rich regions of bacterial subtilisins by combinatorial fragment exchange |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0024319&type=printable |
| work_keys_str_mv | AT ddafyddjones recombininglowhomologyfunctionallyrichregionsofbacterialsubtilisinsbycombinatorialfragmentexchange |