Kinetics and equilibrium for thermolysin-catalyzed syntheses of dipeptide precursors in aqueous/organic biphasic systems

Masamitsu Miyanaga, Michihiro Ohmori, Koreyoshi Imamura, Takaharu Sakiyama, Kazuhiro Nakanishi

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The initial kinetics for the syntheses of N-(benzyloxycarbonyl)-L-alanyl-L-phenylalanine methyl ester (ZAPM) and N-(benzyloxycarbonyl)-L-aspartyl-L-phenylalanine methyl ester (ZDPM) in an aqueous/organic biphasic system, using free thermolysin were elucidated, both experimentally and theoretically. As model organic solvents, ethyl acetate and tert-amyl alcohol were used. The substrate concentration dependencies of the initial rate of syntheses for ZAPM and ZDPM observed in the biphasic system were well simulated using the overall partition coefficients of the substrates and product taking into consideration the effect of the formation of ion-pair complexes between the acid and amine components of the substrate, the initial rate equations determined in an aqueous buffer saturated with the organic solvent, and the pH dependence of the rate constant. The equilibrium yield for the synthesis of ZDPM was also in good agreement with the calculated result using the overall partition coefficients and equilibrium constant measured in the aqueous buffer.

Original languageEnglish
Pages (from-to)43-51
Number of pages9
JournalJournal of Bioscience and Bioengineering
Volume90
Issue number1
DOIs
Publication statusPublished - Jul 2000

Fingerprint

Thermolysin
Dipeptides
aspartyl-phenylalanine
Buffers
Organic solvents
Kinetics
Esters
Substrates
Phenylalanine
Amines
Equilibrium constants
Ions
Acids
Rate constants
Alcohols
phenylalanine methyl ester
alanylphenylalanine
tert-amyl alcohol
ethyl acetate

Keywords

  • Aqueous/organic biphasic reaction
  • Dipeptide precursor
  • Enzymatic synthesis
  • Immobilized enzyme
  • Thermolysin

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering

Cite this

Kinetics and equilibrium for thermolysin-catalyzed syntheses of dipeptide precursors in aqueous/organic biphasic systems. / Miyanaga, Masamitsu; Ohmori, Michihiro; Imamura, Koreyoshi; Sakiyama, Takaharu; Nakanishi, Kazuhiro.

In: Journal of Bioscience and Bioengineering, Vol. 90, No. 1, 07.2000, p. 43-51.

Research output: Contribution to journalArticle

Miyanaga, Masamitsu ; Ohmori, Michihiro ; Imamura, Koreyoshi ; Sakiyama, Takaharu ; Nakanishi, Kazuhiro. / Kinetics and equilibrium for thermolysin-catalyzed syntheses of dipeptide precursors in aqueous/organic biphasic systems. In: Journal of Bioscience and Bioengineering. 2000 ; Vol. 90, No. 1. pp. 43-51.
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