Nakamura Yurie, Matsumoto Takeshi, Nomoto Fumiki, Ueda Mitsuyoshi, Fukuda Hideki, Kondo Akihiko
Department of Chemical Science and Engineering, Faculty of Engineering, Kobe University, Nada-ku, Japan.
Biotechnol Prog. 2006 Jul-Aug;22(4):998-1002. doi: 10.1021/bp060136m.
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 degrees 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.
米根霉脂肪酶（ROL）通过Flo1 N端区域（1100个氨基酸）展示在酿酒酵母的细胞表面，该区域对应一个絮凝功能域。在培养8天后，将酵母细胞于20℃在蒸馏水中孵育8天，展示脂肪酶的酵母全细胞生物催化剂的活性提高了7.3倍。通过孵育发现，存在于细胞壁和细胞内部分的脂肪酶分子数量分别增加了4.5倍和1.8倍，这证明在孵育过程中ROL分子得以表达。具有增强活性的展示ROL的酵母全细胞生物催化剂成功地用于药物前体（R,S）-1-苄氧基-3-氯-2-丙基单琥珀酸酯的光学拆分。此外，它在至少八个反应循环中表现出稳定的活性。这些结果表明，通过在蒸馏水中孵育而具有增强活性的展示ROL的酵母细胞在工业生物转化过程中非常有效。
