Direct Production of D-Arabinose from D-Xylose by a Coupling Reaction Using D-Xylose Isomerase,D-Tagatose 3-Epimerase and D-Arabinose Isomerase

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Klebsiella pneumoniae 40bXX, a mutant strain that constitutively produces D-arabinose isomerase(D-AI), was isolated through a series of repeated subcultures from the parent strain on a mineral salt medium supplemented with L-xylose as the sole carbon source. D-AI could be efficiently immobilized on chitopearl beads. The optimum temperature for the activity of the immobilized enzyme was 40℃ and the enzyme was stable up to 50℃. The D-AI was active at pH 10.0 and was stable in the range of pH 6.0-11.0. The enzyme required manganese ions for maximum activity. Three immobilized enzymes, D-xylose isomerase (D-XI), D-tagatose 3-epimerase (D-TE) and D-AI were used for the preparation of D-arabinose from D-xylose in a coupling reaction. After completion of the reaction, degradation of D-xylulose was carried out by Saccharomyces cerevisiae. The reaction mixture containing D-xylose, D-ribulose and the product was then separated by ion exchange column chromatography. After crystallization, the product was checked by HPLC, IR spectroscopy, NMR spectroscopy and optical rotation measurements. Finally, 2.0g of D-arabinose could be obtained from 5g of the substrate.
社団法人日本生物工学会の論文
2003-04-25