Enhancement of activity of lipase-displaying yeast cells and their application to optical resolution of (R,S)-1-benzyloxy-3-chloro-2-propyl monosuccinate.
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.
Nakamura
Yurie
Y
Department of Chemical Science and Engineering, Faculty of Engineering, Kobe University, Nada-ku, Japan.
Matsumoto
Takeshi
T
Nomoto
Fumiki
F
Ueda
Mitsuyoshi
M
Fukuda
Hideki
H
Kondo
Akihiko
A
D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D004789:Enzyme Activation; D006868:Hydrolysis; D008049:Lipase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015394:Molecular Structure; D012233:Rhizopus / Q000201:enzymology; D012441:Saccharomyces cerevisiae / Q000737:chemistry* / Q000201:enzymology / Q000235:genetics; D013237:Stereoisomerism; D013386:Succinates / Q000737:chemistry* / Q000302:isolation & purification; D013997:Time Factors
C523115:1-benzyloxy-3-chloro-2-propyl monosuccinate; D013386:Succinates; D008049:Lipase