Direct synthesis of Neu5Ac from GlcNAc using NALasc and GlcNAc 2-epimerase

  • 이정규 (서울대학교 응용화학부 유전공학연구소) ;
  • 이정오 (서울대학교 응용화학부 유전공학연구소) ;
  • 이선구 (서울대학교 응용화학부 유전공학연구소) ;
  • 김병기 (서울대학교 응용화학부 유전공학연구소)
  • Published : 2001.11.07

Abstract

GlcNAc 2-epimerase gene from human was cloned. However GIcNAc 2-epimerase was expressed in E. coli as inclusion body formation. Several approaches were tried such as expression in low temperature and low concentration of IPTG. With these treatments production of active form of human GIcNAc 2-epimerase ι ,vas enhanced. For the direct synthesis of NeuAc from GlcNAc and pyruvate, NALase and GlcNAc 2-epimerase were characterized in terms of temperature effect on activity. equilibrium and stability, inhibition by pyruvate etc. For cheap and ease preparation of both the NALase and GlcNAc 2-epimerase, pEN24ma vector was made. which express both the NALasc and GIcNAc 2-epimerase simultaneously. In addition, E. coli BL21(DE3) harboring two plasmids was also made. Of the two systems, the latter was better for the expression of both enzymes.

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