Amino Acids

, Volume 40, Issue 1, pp 215–220 | Cite as

Preparation of optically active β-hydroxy-α-amino acid by immobilized Escherichia coli cells with serine hydroxymethyl transferase activity

  • Gen-Hai Zhao
  • Hui Li
  • Wei Liu
  • Wei-Guo Zhang
  • Fei Zhang
  • Qian Liu
  • Qing-Cai Jiao
Original Article

Abstract

In this research, an improved method for preparation of optically pure β-hydroxy-α-amino acids, catalyzed by serine hydroxymethyl transferase with threonine aldolase activity, is reported. Using recombinant serine hydroxymethyl transferase (SHMT), an enzymatic resolution process was established. A series of new substrates, β-phenylserine, β-(nitrophenyl) serine and β-(methylsulfonylphenyl) serine were used in the resolution process catalyzed by immobilized Escherichia coli cells with SHMT activity. It was observed that the K m for l-threonine was 28-fold higher than that for l-allo-threonine, suggesting that this enzyme can be classified as a low-specificity l-allo-threonine aldolase. The results also shows that SHMT activity with β-phenylserine as substrate was about 1.48-fold and 1.25-fold higher than that with β-(methylsulfonylphenyl) serine and β-(nitrophenyl) serine as substrate, respectively. Reaction conditions were optimized by using 200 mmol/l β-hydroxy-α-amino acid, and 0.1 g/ml of immobilized SHMT cells at pH 7.5 and 45°C. Under these conditions, the immobilized cells were continuously used 10 times, yielding an average conversion rate of 60.4%. Bead activity did not change significantly the first five times they were used, and the average conversion rate during the first five instances was 84.1%. The immobilized cells exhibited favourable operational stability.

Keywords

d-threo-Amino acid Enzymatic resolution Immobilized cells Serine hydroxymethyl transferase 

Notes

Acknowledgments

This work was supported by the National Technology-Innovation Fund (02CJ-13-01-16) and the Open Fund of State Key Laboratory of Pharmaceutical Biotechnology of Nanjing University, P.R. China.

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Gen-Hai Zhao
    • 1
  • Hui Li
    • 1
  • Wei Liu
    • 1
  • Wei-Guo Zhang
    • 1
  • Fei Zhang
    • 1
  • Qian Liu
    • 1
  • Qing-Cai Jiao
    • 1
  1. 1.State Key Laboratory of Pharmaceutical Biotechnology, School of Life ScienceNanjing UniversityNanjingPeople’s Republic of China

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