Home > Publications database > Novel plasmid-free Gluconobacter oxydans strains for production of the natural sweetener 5-ketofructose > print

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005     20220930130230.0
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100 1 _ |a Battling, Svenja
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245 _ _ |a Novel plasmid-free Gluconobacter oxydans strains for production of the natural sweetener 5-ketofructose
260 _ _ |a London
|c 2020
|b Biomed Central
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500 _ _ |a Biotechnologie 1
520 _ _ |a 5-Ketofructose (5-KF) has recently been identified as a promising non-nutritive natural sweetener. Gluconobacter oxydans strains have been developed that allow efficient production of 5-KF from fructose by plasmid-based expression of the fructose dehydrogenase For plasmid-free 5-KF production, we selected four sites in the genome of G. oxydans IK003.1 and inserted the fdhSCL genes under control of the strong P264 promoter into each of these sites. All four recombinant strains expressed fdhSCL and oxidized fructose to 5-KF, but site-specific differences were observed suggesting that the genomic vicinity influenced gene expression. For further improvement, a second copy of the fdhSCL genes under control of P264 was inserted into the second-best insertion site to obtain strain IK003.1::fdhSCL2. The 5-KF production rate and the 5-KF yield obtained with this double-integration strain were considerably higher than for the single integration strains and approached the values of IK003.1 with plasmid-based fdhSCL expression.We identified four sites in the genome of G. oxydans suitable for expression of heterologous genes and constructed a strain with two genomic copies of the fdhSCL genes enabling efficient plasmid-free 5-KF production. This strain will serve as basis for further metabolic engineering strategies aiming at the use of alternative carbon sources for 5-KF production and for bioprocess optimization.
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700 1 _ |a Wohlers, Karen
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700 1 _ |a Igwe, Chika
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700 1 _ |a Kranz, Angela
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700 1 _ |a Pesch, Matthias
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700 1 _ |a Wirtz, Astrid
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700 1 _ |a Baumgart, Meike
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700 1 _ |a Büchs, Jochen
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700 1 _ |a Bott, Michael
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773 _ _ |a 10.1186/s12934-020-01310-7
|g Vol. 19, no. 1, p. 54
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