Enhancement of Activity of Lipase-Displaying Yeast Cells and Their Application to Optical Resolution of (R,S)-1-Benzyloxy-3-Chloro-2-Propyl Monosuccinate

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Abstract
Rhizopus oryzae lipase (ROL) was displayed on the cell surface of Saccharomyces cerevisiae via the Flo1 N‐terminal region (1100 amino acids), which corresponds to a flocculation functional domain. The activity of lipase‐displaying yeast whole‐cell biocatalysts was enhanced 7.3‐fold by incubation of the yeast cells at 20 °C in distilled water for 8 days after 8 day cultivation. The amount of lipase molecules present in cell wall and intracellular fractions was found to be increased 4.5‐ and 1.8‐fold, respectively, by incubation, which proves that ROL molecules are expressed during incubation. The ROL‐displaying yeast whole‐cell biocatalyst with enhanced activity was successfully catalyzed by optical resolution of the pharmaceutical precursor (R,S)‐1‐benzyloxy‐3‐chloro‐2‐propyl monosuccinate. Moreover, it showed stable activity through at least eight reaction cycles. These results demonstrate that ROL‐displaying yeast cells with enhanced activity by incubation in distilled water are very effective in industrial bioconversion processes.

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