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- Title
Identification of sucrose synthase in nonphotosynthetic bacteria and characterization of the recombinant enzymes.
- Authors
Diricks, Margo; De Bruyn, Frederik; Van Daele, Paul; Walmagh, Maarten; Desmet, Tom
- Abstract
Sucrose synthase (SuSy) catalyzes the reversible conversion of sucrose and a nucleoside diphosphate into fructose and nucleotide (NDP)-glucose. To date, only SuSy's from plants and cyanobacteria, both photosynthetic organisms, have been characterized. Here, four prokaryotic SuSy enzymes from the nonphotosynthetic organisms Nitrosomonas Europaea (SuSyNe), Acidithiobacillus caldus (SuSyAc), Denitrovibrio acetiphilus (SusyDa), and Melioribacter roseus (SuSyMr) were recombinantly expressed in Escherichia coli and thoroughly characterized. The purified enzymes were found to display high-temperature optima (up to 80 °C), high activities (up to 125 U/mg), and high thermostability (up to 15 min at 60 °C). Furthermore, SuSyAc, SuSyNe, and SuSyDa showed a clear preference for ADP as nucleotide, as opposed to plant SuSy's which prefer UDP. A structural and mutational analysis was performed to elucidate the difference in NDP preference between eukaryotic and prokaryotic SuSy's. Finally, the physiological relevance of this enzyme specificity is discussed in the context of metabolic pathways and genomic organization.
- Subjects
SUCROSE synthase; RECOMBINANT proteins; PHOTOSYNTHETIC bacteria; NITROSOMONAS europaea; ACIDITHIOBACILLUS caldus; NUCLEOTIDES; FRUCTOSE; BACTERIAL metabolism
- Publication
Applied Microbiology & Biotechnology, 2015, Vol 99, Issue 20, p8465
- ISSN
0175-7598
- Publication type
Article
- DOI
10.1007/s00253-015-6548-7