Thermostable ATP regeneration system using polyphosphate kinase from Thermosynechococcus elongatus BP-1 for d-amino acid dipeptide synthesis

Masaru Sato, Yusuke Masuda, Kohtaro Kirimura, Kuniki Kino

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

d-Alanine-d-alanine ligase from Thermotoga maritima ATCC 43589 (TmDdl) was a useful biocatalyst for synthesizing d-amino acid dipeptides. TmDdl showed a broad substrate specificity at a high temperature; however, ATP was required for its reaction. One of the methods for an effective ATP supply was the coupling reaction with an ATP regeneration system. However, ATP regeneration systems consisted of enzymes from mesophiles and were difficult to operate at high temperatures. Therefore, an ATP regeneration system that could be used at high temperatures was desired to utilize TmDdl for the effective production of d-amino acid dipeptides. To establish a thermostable ATP regeneration system, polyphosphate kinase from a thermophile, Thermosynechococcus elongatus BP-1 (TePpk), was characterized. TePpk showed thermostability up to 70°C; therefore, it was considered that a thermostable ATP regeneration system could be established using TePpk. In the coupling reaction with purified TmDdl and TePpk at 60°C, the amount of ATP required for d-alanyl-d-alanine synthesis could be reduced to 1% of the theoretical amount required when there was no ATP regeneration. When the coupling reaction was applied to a resting cell reaction, ATP was regenerated from an adenosine scaffold in the cell, and d-alanyl-d-alanine was successfully synthesized in the maximum yield of 80% (mol/mol) without the addition of ATP. Thus, an effective synthesis of d-amino acid dipepitides was achieved using the thermostable ATP regeneration system.

Original languageEnglish
Pages (from-to)179-184
Number of pages6
JournalJournal of Bioscience and Bioengineering
Volume103
Issue number2
DOIs
Publication statusPublished - 2007 Feb 1

Keywords

  • ATP regeneration
  • d-alanine-d-alanine ligase
  • d-amino acid dipeptide
  • polyphosphate kinase
  • thermophile

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Fingerprint Dive into the research topics of 'Thermostable ATP regeneration system using polyphosphate kinase from Thermosynechococcus elongatus BP-1 for d-amino acid dipeptide synthesis'. Together they form a unique fingerprint.

  • Cite this