Efficient Bioreduction of Ethyl 4-chloro-3-oxobutanoate to (S)4-chloro-3-hydrobutanoate by Whole Cells of Candida magnoliae in Water/ n-Butyl Acetate Two-phase System

  • Xua Zhinan (Institute of Bioengineering, Department of Chemical and Biochemical Engineering, Zhejiang University) ;
  • Fang Limei (Zhejiang University City College) ;
  • Lin Jianping (Institute of Bioengineering, Department of Chemical and Biochemical Engineering, Zhejiang University) ;
  • Jiang Xiaoxia (Institute of Bioengineering, Department of Chemical and Biochemical Engineering, Zhejiang University) ;
  • Liu Ying (Institute of Bioengineering, Department of Chemical and Biochemical Engineering, Zhejiang University) ;
  • Cen Peilin (Institute of Bioengineering, Department of Chemical and Biochemical Engineering, Zhejiang University)
  • 발행 : 2006.01.01

초록

The asymmetric biosynthesis of ethyl (S)-4-chloro-3-hydrobutanoate from ethyl 4-chloro-3-oxobutanoate was investigated by using whole cells of Candida magnoliae JX120-3 without the addition of glucose dehydrogenase or $NADP^+/NADPH$. In a one-phase system, the bioconversion yield was seriously affected on the addition of 12.1 g/L ethyl 4-chloro-3-oxobutanoate. In order to reduce this substrate inhibition, a water/ n-butyl acetate two-phase system was developed, and the bioreduction conditions optimized with regard to the yield and product enantiometric excess value. The optimal conditions were as following: water to n-butyl acetate volume ratio of 1:1, 4.0 g DCW/L active cells, 50 g/L glucose and $35^{\circ}C$. By adopting a dropwise substrate feeding strategy, high concentration of ethyl 4-chloro-3-oxobutanoate (60 g/L) could be asymmetrically reduced to ethyl (S)-4-chloro-3-hydrobutanoate with high yield (93.8%) and high enantiometric excess value (92.7%).

키워드

참고문헌

  1. Jiang, B., J. F. Liu, and S. Y. Zhao (2001) Enantioselective synthesis for the antipodes of slagenins B and C: establishment of absolute stereochemistry. Org. Lett. 3: 4011-4013 https://doi.org/10.1021/ol016689+
  2. Karanewsky, D. S., M. C. Badia, C. P. Ciosek, Jr., J. A. Robl, M. J. Sofia, L. M. Simpkins, B. DeLange, T. W. Harrity, S. A. Biller, and E. M. Gordon (1990) Phosphorus- containing inhibitors of HMG-CoA reductase. 1. 4- [(2-arylethyl) hydroxyphosphinyl]-3-hydroxy-butanoic acids: a new class of cell-selective inhibitors of cholesterol biosynthesis. J. Med. Chem. 33: 2952-2956 https://doi.org/10.1021/jm00173a007
  3. Kita, K., M. Kataoka, and S. Shimizu (1999) Diversity of 4-chloroacetoacetate ethyl ester-reducing enzymes in yeasts and their application to chiral alcohol synthesis. J. Biosci. Bioeng. 88: 591-598 https://doi.org/10.1016/S1389-1723(00)87085-1
  4. Yasohara, Y., N. Kizaki, J. Hasegawa, S. Takahashi, M. Wada, M. Kataoka, and S. Shimizu (1999) Synthesis of optically active ethyl 4-chloro-3-hydroxybutanoate by microbial reduction. Appl. Microbiol. Biotechnol. 51: 847- 851 https://doi.org/10.1007/s002530051472
  5. Wada, M., M., Kataoka, H. Kawabata, Y. Yasohara, N. Kizaki, J. Hasegawa, and S. Shimizu (1998) Purification and characterization of NADPH-dependent carbonyl reductase, involved in stere-oselective reduction of ethyl 4- chloro-3-oxobuta-noate, from Candida magnoliae. Biosci. Biotechnol. Biochem. 62: 280-285 https://doi.org/10.1271/bbb.62.280
  6. Shimizu, S., M. Kataoka, and K. Kita (1998) Chiral alcohol synthesis with yeast carbonyl reductases. J. Mol. Catal., B Enzym. 5: 321-325 https://doi.org/10.1016/S1381-1177(98)00064-2
  7. Hallinan, K. O., D. H. G. Crout, J. R. Hunt, A. S. Carter, H. Dalton, R. A. Holt, and J. Crosby (1995) Yeast catalysed reduction of $\beta$-keto esters. II. Optimization of the stereospecific reduction by Zygosaccharomyces rouxii. Biocatal. Biotransformation 12: 179-191 https://doi.org/10.3109/10242429508998161
  8. Patel, R. N., C. G. McNamee, A. Banerjee, J. M. Howell, R. S. Robinson, and L. Szarka (1992) Stereoselective reduction of $\beta$-keto esters by Geotrichum candidum. Enzyme Microb. Technol. 14: 731-738 https://doi.org/10.1016/0141-0229(92)90113-3
  9. Liu, Y., Z. Xu, K. Jing, et al. (2004) A novel NADPHregenerating system for production of chiral alcohols. The 8th China-Japan-Korea Joint Symposium on Enzyme Engineering. pp. 105. Hangzhou, China
  10. Liu, Y., Z. Xu, K. Jing, X. Jiang, J. Lin, F. Wang, and P. Cen (2005) Asymmetric reduction of COBE to CHBE with two co-existing, recombinant Escherichia coli strains. Biotechnol. Lett. 27: 119-125 https://doi.org/10.1007/s10529-004-7336-0
  11. Lou, W., M. Zong, Y. Zhang, and H. Wu (2004) Efficient synthesis of optically active organosilyl alcohol via asymmetric reduction of acyl silane with immobilized yeast. Enzyme Microb. Technol. 35: 190-196 https://doi.org/10.1016/j.enzmictec.2004.04.009
  12. Bar, R. (1988) Effect of interphase mixing on a waterorganic solvent two-liquid phase microbial system: ethanol fermentation. J. Chem. Technol. Biotechnol. 43: 49-62
  13. Shimizu, S., M. Kataoka, M. Katoh, T. Morikawa, T. Miyoshi, and H. Yamada (1990) Stereoselective reduction of ethyl 4-chloro-3-oxobutanoate by a microbial aldehyde reduction in an organic solvent-water diphasic system. Appl. Environ. Microbiol. 56: 2374-2377
  14. Hocknull, M. D. and M. D. Lilly (1988) Stability of the steroid delta-dehydrogenation system of Artehrobacter simplex in organic solvent-water two liquid phase environments. Enzyme Microb. Technol. 10: 669-674 https://doi.org/10.1016/0141-0229(88)90058-0
  15. Darrugo, P., G. P. Fantoni, S. Servi, et al. (1997) The effect of absorbing on substrate concentration and enantiomeric excess in yeast reduction. Tetrahedron Asymmetry 14: 375-379