Abstract
Objective
To characterize l-rhamnose isomerase (l-RI) from the thermophilic bacterium Clostridium stercorarium and apply it to produce d-allose from d-allulose.
Results
A recombinant l-RI from C. stercorarium exhibited the highest specific activity and catalytic efficiency (k cat/K m) for l-rhamnose among the reported l-RIs. The l-RI was applied to the high-level production of d-allose from d-allulose. The isomerization activity for d-allulose was maximal at pH 7, 75 °C, and 1 mM Mn2+ over 10 min reaction time. The half-lives of the l-RI at 65, 70, 75, and 80 °C were 22.8, 9.5, 1.9, and 0.2 h, respectively. To ensure full stability during 2.5 h incubation, the optimal temperature was set at 70 °C. Under the optimized conditions of pH 7, 70 °C, 1 mM Mn2+, 27 U l-RI l−1, and 600 g d-allulose l−1, l-RI from C. stercorarium produced 199 g d-allose l−1 without by-products over 2.5 h, with a conversion yield of 33% and a productivity of 79.6 g l−1 h−1.
Conclusion
To the best of our knowledge, this is the highest concentration and productivity of d-allose reported thus far.
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Acknowledgements
This work was supported by a grant from the Next-Generation BioGreen 21 Program (No. PJ01113701), Rural Development Administration, Republic of Korea.
Supporting information
Supplementary Fig. 1—Effect of metal ions on the production of d-allose from d-allulose by l-RI from C. stercorarium.
Supplementary Fig. 2—Effect of pH on the production of d-allose from d-allulose by l-RI from C. stercorarium.
Supplementary Fig. 3—Effect of temperature on the production of d-allose from d-allulose by l-RI from C. stercorarium.
Supplementary Fig. 4—Effect of substrate concentration on the production of d-allose from d-allulose by l-RI from C. stercorarium.
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Seo, MJ., Choi, JH., Kang, SH. et al. Characterization of l-rhamnose isomerase from Clostridium stercorarium and its application to the production of d-allose from d-allulose (d-psicose). Biotechnol Lett 40, 325–334 (2018). https://doi.org/10.1007/s10529-017-2468-1
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DOI: https://doi.org/10.1007/s10529-017-2468-1