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<span style="font-variant: small-caps;">d</span>-pantolactone is an intermediate in the synthesis of <span style="font-variant: small-caps;">d</span>-pantothenic acid, which is known as vitamin B₅. The commercial synthesis of <span style="font-variant: small-caps;">d</span>-pantolactone is carried out through the selective resolution of <span style="font-variant: small-caps;">dl</span>-pantolactone catalyzed by lactone hydrolase. In contrast to a kinetic resolution approach, the deracemization of <span style="font-variant: small-caps;">dl</span>-pantolactone is a simpler, greener, and more sustainable way to obtain <span style="font-variant: small-caps;">d</span>-pantolactone with high optical purity. Herein, an efficient three-enzyme cascade was developed for the deracemization of <span style="font-variant: small-caps;">dl</span>-pantolactone, using <span style="font-variant: small-caps;">l</span>-pantolactone dehydrogenase from <i>Amycolatopsis methanolica</i> (<i>Ame</i>LPLDH), conjugated polyketone reductase from <i>Zygosaccharomyces parabailii</i> (<i>Zpa</i>CPR), and glucose dehydrogenase from <i>Bacillus subtilis</i> (<i>Bs</i>GDH). The <i>Ame</i>LPLDH was used to catalyze the dehydrogenated <span style="font-variant: small-caps;">l</span>-pantolactone into ketopantolactone; the <i>Zpa</i>CPR was used to further catalyze the ketopantolactone into <span style="font-variant: small-caps;">d</span>-pantolactone; and glucose dehydrogenase together with glucose fulfilled the function of coenzyme regeneration. All three enzymes were co-expressed in <i>E. coli</i> strain BL21(DE3), which served as the whole-cell biocatalyst. Under optimized conditions, 36 h deracemization of 1.25 M <span style="font-variant: small-caps;">dl</span>-pantolactone <span style="font-variant: small-caps;">d</span>-pantolactone led to an <i>e.e.</i>_p value of 98.6%, corresponding to productivity of 107.7 g/(l·d).