Biotechnology Bulletin ›› 2019, Vol. 35 ›› Issue (6): 221-226.doi: 10.13560/j.cnki.biotech.bull.1985.2018-0977

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Construction of ATP Synthetic Strain and Its Application in the Production of S- adenosylmethionine

JIANG Lin-lin, WU Lei, XU Hai-xia, HUANG Jian-li, ZHANG Yong-jin, XU Qi, YANG Yong   

  1. Zhejiang Hisun Pharmaceutical Co. Ltd.,Taizhou 318000
  • Received:2018-11-13 Online:2019-06-26 Published:2019-07-08

Abstract: In order to achieve ATP production as well as reduce the cost of producing S-adenosylmethionine,a recombinant Escherichia coli strain expressing adenosine kinase,adenylate kinase and acetate kinase simultaneously was constructed for ATP synthesis. The conditions of transforming ATP were optimized,and the optimized reaction system was as:adenosine 30 mmol/L,acetylphosphate dilithium 135 mmol/L,magnesium sulfate 5 mmol/L,borax 50 mmol/L,E. coli strain 2 g/L(wet weight),initial pH 7.5,the reaction temperature 35℃,and reacting for 3 h,the conversion rate was over 99%. Based on the above result,the reaction system was enlarged in 5 L. After reaction ended,50 g/L(wet weight)recombinant E. coli strain expressing adenosylmethionine synthase,65 mmol/L D,L-methionine and 15 mmol/L magnesium sulfate were added to the reaction system,the concentration of S-adenosylmethionine was up to 8.7 g/L and the conversion rate reached 72.5% after 18 h transformation.

Key words: ATP synthesis, adenosine kinase, adenylate kinase, acetate kinase, S-adenosylmethionine