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Two groups of thermophilic amino acid aminotransferases exhibiting broad substrate specificities for the synthesis of phenylglycine derivatives

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Abstract

Thirty two thermophilic amino acid aminotransferases (AATs) were expressed in Escherichia coli as soluble and active proteins. Based on their primary structures, the 32 AATs were divided into four phylogenetic groups (classes I, II, IV, and V). The substrate specificities of these AATs were examined, and 12 AATs were found capable of synthesizing ring-substituted phenylglycine derivatives such as hydroxyl-, methoxy-, and fluorophenylglycines. Eleven out of the 12 AATs were enzymes belonging to two phylogenetic groups namely, one subgroup of the class I family and the class IV family. AATs in these two groups may thus be useful for the synthesis of a variety of ring-substituted phenylglycine derivatives.

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Acknowledgments

We thank professor Isao Shimizu, Consolidated Research Institute for Advanced Science and Medical Care, Waseda University, for providing us the keto acids and Ms. Ayako Hashimoto for technical assistance. This research work was done in part at the “Center for Practical Chemical Wisdom” supported by Global COE program of Ministry of Education, Culture, Sports, Science and Technology.

Author information

Correspondence to Daisuke Koma.

Electronic supplementary material

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Supplemental Fig. S1

SDS-PAGE analysis of the expression and purification of MJ1008, TTE0933, and TTHA0124. The arrows indicate the respective desired proteins, and CFE and PF mean cell-free extract and purified enzyme, respectively (PPT 335 kb)

Supplemental Table S1

Primer sequences for the cloning of the aminotransferase genes (DOC 94.5 kb)

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Koma, D., Sawai, T., Hara, R. et al. Two groups of thermophilic amino acid aminotransferases exhibiting broad substrate specificities for the synthesis of phenylglycine derivatives. Appl Microbiol Biotechnol 79, 775–784 (2008). https://doi.org/10.1007/s00253-008-1487-1

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Keywords

  • Aminotransferase
  • Thermophile
  • Phenylglycine
  • Unnatural amino acid
  • Nonproteinogenic amino acid