Efficient Allitol Bioproduction from d -Fructose Catalyzed by Recombinant E. coli Whole Cells, and the Condition Optimiz
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Efficient Allitol Bioproduction from D-Fructose Catalyzed by Recombinant E. coli Whole Cells, and the Condition Optimization, Product Purification Xin Wen 1 & Huibin Lin 2 & Yilin Ren 3 Jianqun Lin 1 & Jianqiang Lin 1
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& Can Li & Chengjia Zhang & Xin Song &
Received: 27 December 2019 / Accepted: 22 May 2020/ # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract
Allitol is a kind of rare sugar alcohol with potential application value. An engineered strain, which simultaneously expressed D-psicose-3-epimerase (DPE), ribitol dehydrogenase (RDH), and formate dehydrogenase (FDH) three enzymes, was constructed by cloning above three genes into one plasmid and transformed into the host E. coli strain, and used as the whole-cell catalysts for biotransformation of allitol from the low-cost substrate of D-fructose. The whole cell allitol biotransformation conditions were optimized. The medium, recombinant gene induction conditions, and the substrate feeding rate for cultivation of the catalytic cells were optimized. Then, the fed-batch culture was made and scaled up to 10 L fermentor. Finally, 63.44 g/L allitol was obtained from 100 g/ L D-fructose after 3 h of biotransformation, and the allitol crystals of 99.9% purity were obtained by using cooling recrystallization. The allitol production method developed in this research has high product purity, and is highly efficient, easily scaled up, and suitable for large-scale production of highly purified allitol. Keywords Allitol . D-fructose . Whole-cell biotransformation . Fed-batch culture . Cooling crystallization
Introduction Allitol is an important rare sugar alcohol, which not only has significant application value in pharmaceutical and food industries [1–4] but also is vital for production of other rare sugars. As it is in the center between the D and L-type mono-sugars, it can be used to synthesize L-type rare sugars like L-psicose and L-fructose, suggested by Izumoring strategy [5–7]. * Xin Song [email protected] * Jianqiang Lin [email protected] Extended author information available on the last page of the article
Applied Biochemistry and Biotechnology
Although allitol can be produced by extraction from natural resources or by chemical synthesis, the yield and purity of these methods were very low [4, 8–10]. Besides, these methods were not environmental unfriendly. In contrast, the biotransformation method has the advantages of high purity, no toxic byproducts, mild reaction conditions, low cost, and environmental friendly, which is the best choice for allitol production. The allitol biotransformation scheme is shown in Fig. 1a. Allitol could be produced from D-psicose or Dfructose by enzymatic catalysis by using D-psicose-3-epimerase (DPE) or D-tagatose-3epimerase (DTE), ribitol dehydrogenase (RDH), and formate dehydrogenase (FDH) [1, 11–14]. However, enzyme extraction and purification were cumbersome and of high cost. In addition, initial addition of the coenzyme of NADH was needed for the enzymatic catalysis method, which
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