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Screening, characterization and utilization of D-amino acid aminotransferase to obtain D-phenylalanine

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Abstract

D-phenylalanine (D-Phe) is an important side-chain building block for semi-synthetic penicillins and cephalosporins. To synthesize D-Phe, D-amino acid aminotransferases (Dat) from Bacillus subtilis WB600, Bacillus licheniformis ATCC 14580, and Bacillus amyloliquefaciens were firstly compared. The theoretical 3 dimensional models of Dat were constructed, which showed appropriate configuration for synthesis of D-Phe in virtual screening. The comparison 3 different Dat on D-Phe formation was tested. All the heterogenous Dat showed high thermostability and pH stability, while Dat from B. subtilis demonstrated a better potential for Phe production than those from B. licheniformis and B. amyloliquefaciens. Then the dat from B. subtilis was expressed in an L-Phe producing chassis of Escherichia coli W14 (pR15ABK) to obtain D-Phe and different factors affecting D-Phe accumulation were tested. The engineered strain E. coli BCEA (pR15ABKApRdatBS) accumulated 1.72 g/L D-Phe in a 15-L jar fermenter which is the highest fermentation concentration that had been reported.

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Authors and Affiliations

  1. The Key Laboratory of Industrial Biotechnology of Ministry of Education, National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi, 214122, China

    R. X. Liu, S. P. Liu, S. Cheng, L. Zhang, Z. Y. Ding, Z. H. Gu & G. Y. Shi

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  1. R. X. Liu
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  2. S. P. Liu
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  3. S. Cheng
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  4. L. Zhang
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  5. Z. Y. Ding
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  6. Z. H. Gu
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  7. G. Y. Shi
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Correspondence to R. X. Liu.

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Liu, R.X., Liu, S.P., Cheng, S. et al. Screening, characterization and utilization of D-amino acid aminotransferase to obtain D-phenylalanine. Appl Biochem Microbiol 51, 695–703 (2015). https://doi.org/10.1134/S0003683815060095

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  • DOI: https://doi.org/10.1134/S0003683815060095

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