Engineered cyclodextrin glucanotransferases from Bacillus sp. G‐825‐6 produce large‐ring cyclodextrins with high specificity
Abstract Cyclodextrin glucanotransferases (CGTases) synthesize cyclic oligosaccharides (cyclodextrins, CD) from starch. A CGTase from Bacillus sp. G‐825‐6 was engineered by site‐directed mutagenesis at two positions by the construction of the variants Y183W, Y183R, D358R, Y183W/D358R, and Y183R/D358...
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doaj-a2c22178f13d4257acf96176e39ec7682020-11-24T22:04:00ZengWileyMicrobiologyOpen2045-88272019-06-0186n/an/a10.1002/mbo3.757Engineered cyclodextrin glucanotransferases from Bacillus sp. G‐825‐6 produce large‐ring cyclodextrins with high specificityChristian Sonnendecker0Susanne Melzer1Wolfgang Zimmermann2Department of Microbiology and Bioprocess Technology, Institute of Biochemistry Leipzig University Leipzig GermanyDepartment of Microbiology and Bioprocess Technology, Institute of Biochemistry Leipzig University Leipzig GermanyDepartment of Microbiology and Bioprocess Technology, Institute of Biochemistry Leipzig University Leipzig GermanyAbstract Cyclodextrin glucanotransferases (CGTases) synthesize cyclic oligosaccharides (cyclodextrins, CD) from starch. A CGTase from Bacillus sp. G‐825‐6 was engineered by site‐directed mutagenesis at two positions by the construction of the variants Y183W, Y183R, D358R, Y183W/D358R, and Y183R/D358R. Among CD composed of 7–12 glucose units (CD7–CD12), Y183W mainly produced CD8. Y183R had completely lost its ability to synthesize CD7, and CD8 and the larger CD were the only cyclic oligosaccharides produced. D358R also formed mainly CD8–CD12 during a reaction time of 24 hr. The double mutant Y183W/D358R showed combined characteristics of the single mutations with very low CD7 cyclization activity and an increased formation of the larger CD. The results show that CGTases synthesizing mainly CD8–CD12 can be constructed allowing a convenient production of larger CD in significant amounts as host molecules in supramolecular complexing reactions.https://doi.org/10.1002/mbo3.757Bacilluscyclodextrin glucanotransferasecyclodextrinsprotein engineeringside‐directed mutagenesis |
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DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Christian Sonnendecker Susanne Melzer Wolfgang Zimmermann |
spellingShingle |
Christian Sonnendecker Susanne Melzer Wolfgang Zimmermann Engineered cyclodextrin glucanotransferases from Bacillus sp. G‐825‐6 produce large‐ring cyclodextrins with high specificity MicrobiologyOpen Bacillus cyclodextrin glucanotransferase cyclodextrins protein engineering side‐directed mutagenesis |
author_facet |
Christian Sonnendecker Susanne Melzer Wolfgang Zimmermann |
author_sort |
Christian Sonnendecker |
title |
Engineered cyclodextrin glucanotransferases from Bacillus sp. G‐825‐6 produce large‐ring cyclodextrins with high specificity |
title_short |
Engineered cyclodextrin glucanotransferases from Bacillus sp. G‐825‐6 produce large‐ring cyclodextrins with high specificity |
title_full |
Engineered cyclodextrin glucanotransferases from Bacillus sp. G‐825‐6 produce large‐ring cyclodextrins with high specificity |
title_fullStr |
Engineered cyclodextrin glucanotransferases from Bacillus sp. G‐825‐6 produce large‐ring cyclodextrins with high specificity |
title_full_unstemmed |
Engineered cyclodextrin glucanotransferases from Bacillus sp. G‐825‐6 produce large‐ring cyclodextrins with high specificity |
title_sort |
engineered cyclodextrin glucanotransferases from bacillus sp. g‐825‐6 produce large‐ring cyclodextrins with high specificity |
publisher |
Wiley |
series |
MicrobiologyOpen |
issn |
2045-8827 |
publishDate |
2019-06-01 |
description |
Abstract Cyclodextrin glucanotransferases (CGTases) synthesize cyclic oligosaccharides (cyclodextrins, CD) from starch. A CGTase from Bacillus sp. G‐825‐6 was engineered by site‐directed mutagenesis at two positions by the construction of the variants Y183W, Y183R, D358R, Y183W/D358R, and Y183R/D358R. Among CD composed of 7–12 glucose units (CD7–CD12), Y183W mainly produced CD8. Y183R had completely lost its ability to synthesize CD7, and CD8 and the larger CD were the only cyclic oligosaccharides produced. D358R also formed mainly CD8–CD12 during a reaction time of 24 hr. The double mutant Y183W/D358R showed combined characteristics of the single mutations with very low CD7 cyclization activity and an increased formation of the larger CD. The results show that CGTases synthesizing mainly CD8–CD12 can be constructed allowing a convenient production of larger CD in significant amounts as host molecules in supramolecular complexing reactions. |
topic |
Bacillus cyclodextrin glucanotransferase cyclodextrins protein engineering side‐directed mutagenesis |
url |
https://doi.org/10.1002/mbo3.757 |
work_keys_str_mv |
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