Inulin and levan synthesis by probiotic Lactobacillus gasseri strains: characterization of three novel fructansucrase enzymes and their fructan products
| Autor: | Slavko Kralj, Marc J. E. C. van der Maarel, Hans Leemhuis, Munir A. Anwar, Lubbert Dijkhuizen, Anna Villar Pique |
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| Přispěvatelé: | Groningen Biomolecular Sciences and Biotechnology, Host-Microbe Interactions, Molecular Microbiology |
| Rok vydání: | 2010 |
| Předmět: |
Glycoside Hydrolases
Inulin Molecular Sequence Data Lactobacillus gasseri LACTIC-ACID BACTERIA Microbiology law.invention Substrate Specificity chemistry.chemical_compound Fructan FUNCTIONAL-CHARACTERIZATION Bacterial Proteins law INULOSUCRASE Raffinose Cloning Molecular SUCROSE chemistry.chemical_classification Inulosucrase LEVANSUCRASE biology REUTERI Probiotics Levansucrase GENOME SEQUENCE Oligosaccharide biology.organism_classification STREPTOCOCCUS-MUTANS Fructans Kinetics Lactobacillus FRUCTOSYLTRANSFERASE chemistry Biochemistry Recombinant DNA JOHNSONII NCC-533 |
| Zdroj: | Microbiology-Sgm, 156(4), 1264-1274. MAIK NAUKA/INTERPERIODICA/SPRINGER |
| ISSN: | 1465-2080 1350-0872 |
| Popis: | Fructansucrase enzymes polymerize the fructose moiety of sucrose into levan or inulin fructans, with β(2-6) and β(2-1) linkages, respectively. Here, we report an evaluation of fructan synthesis in three Lactobacillus gasseri strains, identification of the fructansucrase-encoding genes and characterization of the recombinant proteins and fructan (oligosaccharide) products. High-performance anion-exchange chromatography and nuclear magnetic resonance analysis of the fructo-oligosaccharides (FOS) and polymers produced by the L. gasseri strains and the recombinant enzymes revealed that, in situ, L. gasseri strains DSM 20604 and 20077 synthesize inulin (and oligosaccharides) and levan products, respectively. L. gasseri DSM 20604 is only the second Lactobacillus strain shown to produce inulin polymer and FOS in situ, and is unique in its distribution of FOS synthesized, ranging from DP2 to DP13. The probiotic bacterium L. gasseri DSM 20243 did not produce any fructan, although we identified a fructansucrase-encoding gene in its genome sequence. Further studies showed that this L. gasseri DSM 20243 gene was prematurely terminated by a stop codon. Exchanging the stop codon for a glutamine codon resulted in a recombinant enzyme producing inulin and FOS. The three recombinant fructansucrase enzymes characterized from three different L. gasseri strains have very similar primary protein structures, yet synthesize different fructan products. An interesting feature of the L. gasseri strains is that they were unable to ferment raffinose, whereas their respective recombinant enzymes converted raffinose into fructan and FOS. |
| Databáze: | OpenAIRE |
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