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Tropical Journal of Pharmaceutical Research
Pharmacotherapy Group, Faculty of Pharmacy, University of Benin, Benin City, Nigeria
ISSN: 1596-5996
EISSN: 1596-9827
Vol. 15, No. 12, 2016, pp. 2701-2708
Bioline Code: pr16355
Full paper language: English
Document type: Research Article
Document available free of charge

Tropical Journal of Pharmaceutical Research, Vol. 15, No. 12, 2016, pp. 2701-2708

 en Synthesis of allitol from D-psicose using ribitol dehydrogenase and formate dehydrogenase
Hassanin, Hinawi AM; Letsididi, Rebaone; Koko, Marwa YF; Mu, Wanmeng; Elferga, Ammar & Jiang, Bo

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 check for this species in other resources 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, LC-MS, 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

 
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