Co-immobilised aspartase and transaminase for high-yield synthesis of l-phenylalanine

Max Cárdenas-Fernández, Elvira Khalikova, Timo Korpela, Carmen López, Gregorio Álvaro

Research output: Contribution to journalArticleResearchpeer-review

6 Citations (Scopus)

Abstract

© 2014 Elsevier B.V. l-phenylalanine (Phe) was synthesised by coupling the enzymes aspartase (AspB) catalysing the synthesis of l-aspartate from fumarate and NH4Cl and microbial aspartate transaminase (TA) catalysing the transfer of the amino group from l-aspartate to phenylpyruvate. Phe synthesis was studied with enzymes in solution and immobilised separately and together on amino-epoxy Relizyme® support. Immobilisation efficiencies and recovered activities of co-immobilised enzymes were slightly lower than those obtained when immobilised separately. Substrate and enzyme concentrations for the synthesis reactions were optimised as follows: co-immobilised 0.3U/mL AspB and 2U/mL TA, 0.15M fumarate, 0.3M NH4Cl, 0.1M phenylpyruvate, 0.1mM pyridoxal-5'-phosphate (PLP) at pH 7.5 and 37°C. Total reaction yield of 83% and Phe yield of 95% were obtained. The initial rates of the reactions catalysed by co-immobilised enzymes were similar to those obtained when the reactions were catalysed by free enzymes, indicating negligible diffusional limitations associated to the application of the co-immobilised enzymes.
Original languageEnglish
Pages (from-to)173-178
JournalBiochemical Engineering Journal
Volume93
DOIs
Publication statusPublished - 5 Jan 2015

Keywords

  • Aspartase
  • Co-immobilised enzymes
  • Immobilisation on epoxy support
  • L-phenylalanine
  • One-pot multienzymatic reaction
  • Transaminase

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