Article Sidebar
Published:
Jan 13, 2017Keywords:
Article Details
Submission of a manuscript to this journal is a representation that the manuscript has not been published previously and is not under consideration for publication elsewhere.
All authors named in each manuscript would be required to sign a form (to be supplied by the Editor) so that they may retain their copyright in the article but to assign to us (the Publishers) and its licensees in perpetuity, in all forms, formats and media (whether known or created in the future) to (i) publish, reproduce, distribute, display and store the contribution, (ii) translate the contribution into other languages, create adaptations, reprints, include within collections and create summaries, extracts and/or abstracts of the contribution, (iii) create any other derivative works(s) based on the contribution, (iv) to exploit all subsidiary rights in the contribution, (v) the inclusion of electronic links from the contribution to third party material where-ever it may be located, and (vi) license any thrid party to do any or all of the above.
Main Article Content
Synthesis of allitol from D-psicose using ribitol dehydrogenase and formate dehydrogenase
Abstract
Purpose: To synthesize allitol from D-psicose by a combination of novel ribitol dehydrogenase (RDH) and formate dehydrogenase (FDH) under optimised production conditions.
Methods: RDH and FDH genes were cloned and introduced into pET-22b(+) vectors for expression in Escherichia coli to produce the corresponding enzymes. The effects of temperature, pH, shaking velocity (75, 100, 125, and 150 rpm), and shaking type (horizontal and vortex) were optimised to maximise the production yield of allitol. The final product was purified and subjected to nuclear magnetic resonance (NMR) spectroscopy, infrared (IR) spectrometry, and liquid chromatography-mass spectrometry (LC-MS) to confirm its structure.
Results: The optimal pH and temperature for the reaction were 7.5 and 40 °C, respectively. The results revealed that allitol yield significantly increased with increase in reaction shaking velocity and reached a maximum yield of 95.60 ± 0.54 % at 150 rpm shaking velocity after 6 h of reaction. When the reaction was run under horizontal shaking, allitol yield increased from 100.00 ± 6.05 (without shaking) to 124.20 ± 9.70 %. Twenty milligrams of D-psicose were successfully reduced to allitol under optimum conditions with a high production yield of 16.7 ± 0.62 mg after 6 h. No by-products were formed during or after the reaction. The produced allitol had a purity of 95 %, and its structure was confirmed by HPLC, IR, LCMS, and NMR spectral analyses.
Conclusion: Using D-psicose as a substrate, allitol with 95 % purity was successfully produced by the combination of novel RDH and FDH.
Keywords: Allitol, Ribitol dehydrogenase, Formate dehydrogenase, D-Psicose, Providencia alcalifaciens
