Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Metabolic Engineering of Corynebacterium glutamicum for N-acetylglucosamine Production

N-아세틸글루코사민 생산을 위한 코리네박테리움 글루타미컴의 대사공학

  • Kim, Jin-Yeon (Department of Pharmacy, Kyungsung University) ;
  • Kim, Bu-yeon (Major in Food Biotechnology, School of Food Biotechnology & Nutrition, Kyungsung University) ;
  • Moon, Kyung-Ho (Department of Pharmacy, Kyungsung University) ;
  • Lee, Jin-Ho (Major in Food Biotechnology, School of Food Biotechnology & Nutrition, Kyungsung University)
  • Received : 2018.12.25
  • Accepted : 2019.01.04
  • Published : 2019.03.28
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Abstract

Recombinant Corynebacterium glutamicum producing N-acetylglucosamine (GlcNAc) was constructed by metabolic engineering. To construct a basal strain producing GlcNAc, the genes nagA, nagB, and nanE encoding N-acetylglucosamine-6-phosphate deacetylase, glucosamine-6-phosphate deaminase, and N-acetylmannosamine-6-phosphate epimerase, respectively, were sequentially deleted from C. glutamicum ATCC 13032, yielding strain KG208. In addition, the genes glmS and gna1 encoding glucosamine-6-phosphate synthase and glucosamine-6-phosphate N-acetyltransferase, which originated from C. glutamicum and Saccharomyces cerevisiae, respectively, were expressed in several expression vectors. Among several combinations of glmS and gna1 expression, recombinant cells expressing glmS and gna1 under control of the ilvC promoter produced 1.77 g/l of GlcNAc and 0.63 g/l of glucosamine in flask cultures.

대사공학을 이용하여 N-아세틸글루코사민(GlcNAc)을 생산하는 재조합 Corynebacterium glutamicum을 개발하였다. 먼저 GlcNAc를 생산하는 기반균주를 제작하기 위하여, N-acetylglucosamine-6-phosphate deacetylase와 glucosamine-6-phosphate deaminase를 암호화하는 nagAB와 N-acetylmannosamine-6-phosphate epimerase를 암호화하는 nanE를 C. glutamicum ATCC 13032에서 순차적으로 결손하여, 최종적으로 KG208 균주를 제작하였다. 또한, glucosamine-6-phosphate synthase를 암호화하는 C. glutamicum 유래의 glmS와 glucosamine-6-phosphate N-acetyltransferase를 암호화하는 Saccharomyces cerevisiae 유래의 gna1을 각각 여러 발현벡터에 클로닝하였다. 여러 발현 조합의 플라스미드들 중에서 pCXI40-glmS와 pCEI40-gna1을 함유한 제조합균주 KG440은 삼각플라스크 발효에서 1.77 g/l의 GlcNAc와 0.63 g/l의 글루코사민을 생산하였다.

Keywords

References

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