Effect of loop-grafting on the activity, stability and regioselectivity of Priestia megaterium levansucrase using two inulosucrases as loop donors Article Swipe
Maria Elena Ortiz‐Soto
,
K.W. Schmid
,
C. Weber
,
Martin Gross
,
Jürgen Seibel
·
YOU?
·
· 2025
· Open Access
·
· DOI: https://doi.org/10.1016/j.ijbiomac.2025.141584
YOU?
·
· 2025
· Open Access
·
· DOI: https://doi.org/10.1016/j.ijbiomac.2025.141584
Active-site loops of enzymes modulate activity, stability, regioselectivity and substrate specificity. This study examines the impact of loop transplantation on Priestia megaterium levansucrase Pm-SacB, which produces levan-type, β-(2,6)-linked fructooligosaccharides. Inulosucrases from Lactobacillus johnsonii (Lj-InuJ) and Halalkalicoccus jeotgali (Hje-Inu), which synthesize β-(2,1)-linked inulin fructans, served as donors of eight active-site loops. Most Lj-InuJ-based loop-grafting enzymes produce smaller oligosaccharides than Pm-SacB, and variants carrying a Lj-InuJ loop 3, whether alone or in combination with grafted-loops 7 and 8, exhibit inulin-type oligosaccharides 1-kestose and 1-nystose among their products. Pm-SacB does not synthesize inulin-type oligosaccharides other than 1-kestose but can elongate them via β-(2,6)-linkages. A construct containing Lj-InuJ loops 3, 7 and 8, along with the mutations S372N, K373R and H423Y (variant Lj-Loop3-7-8**), has impaired 6-kestose and levan synthesis, accumulates 14-fold more 1-kestose than Pm-SacB and outperforms other variants in the synthesis of 1-nystose. The enzyme exhibits similar thermal stability to Pm-SacB, but its catalytic efficiency is sevenfold lower. The product profile of Lj-Loop3-7-8** could not be replicated by mutating reported levan- and inulin-binding residues in Pm-SacB, suggesting that modifying neighboring residues, in addition to those involved in FOS-binding may be necessary to gain access to levansucrases with unique properties.
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- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1016/j.ijbiomac.2025.141584
- OA Status
- hybrid
- Cited By
- 2
- References
- 35
- Related Works
- 10
- OpenAlex ID
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All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W4408016688Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.1016/j.ijbiomac.2025.141584Digital Object Identifier
- Title
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Effect of loop-grafting on the activity, stability and regioselectivity of Priestia megaterium levansucrase using two inulosucrases as loop donorsWork title
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articleOpenAlex work type
- Language
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enPrimary language
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2025Year of publication
- Publication date
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2025-02-27Full publication date if available
- Authors
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Maria Elena Ortiz‐Soto, K.W. Schmid, C. Weber, Martin Gross, Jürgen SeibelList of authors in order
- Landing page
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https://doi.org/10.1016/j.ijbiomac.2025.141584Publisher landing page
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YesWhether a free full text is available
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hybridOpen access status per OpenAlex
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https://doi.org/10.1016/j.ijbiomac.2025.141584Direct OA link when available
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Levansucrase, Loop (graph theory), Regioselectivity, Bacillus megaterium, Chemistry, Grafting, Biochemistry, Biology, Organic chemistry, Mathematics, Catalysis, Genetics, Combinatorics, Bacteria, Polymer, Bacillus subtilisTop concepts (fields/topics) attached by OpenAlex
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2Total citation count in OpenAlex
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2025: 2Per-year citation counts (last 5 years)
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10Other works algorithmically related by OpenAlex
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