Surfactant-protease complex as a novel biocatalyst for peptide synthesis in hydrophilic organic solvents

Shin ya Okazaki, Masahiro Goto, Shintaro Furusaki

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

The peptide synthesis from N-acetyl-L-phenylalanine ethyl ester with alaninamide catalyzed by a surfactant-protease complex has been performed in anhydrous hydrophilic organic solvents. Proteases derived from various sources were converted to surfactant-coated complexes with a nonionic surfactant. The surfactant-subtilisin Carlsberg (STC) complex had a higher enzymatic activity than the other protease complexes and the initial reaction rate in tert-amyl alcohol was 26-fold that of STC lyophilized from an optimum aqueous buffer solution. Native STC hardly catalyzed the same reaction. The addition of water to the reaction medium activated the lyophilized STC, however, the reaction rate was much lower than that of the STC complex, and a hydrolysis reaction preferentially proceeded. The STC complex exhibited a high catalytic activity in hydrophilic organic solvents (e.g. tertiary alcohol). The addition of dimethylformamide as a cosolvent improved the solubility of amino acid amides and further activated the STC complex due to the water mimicking effect. When hydrophilic amino acid amides were employed as an acyl acceptor, the peptide formation proceeded efficiently compared to that using hydrophobic substrates. The surfactant-STC complex is a powerful biocatalyst for peptide synthesis because the STC complexes display a high catalytic activity in anhydrous hydrophilic organic solvents and did not require the excess amount of water. Thus the side (hydrolysis) reaction is effectively suppressed and the yield in the dipeptide formation is considerably high.

Original languageEnglish
Pages (from-to)159-164
Number of pages6
JournalEnzyme and Microbial Technology
Volume26
Issue number2-4
DOIs
Publication statusPublished - Jan 1 2000

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Subtilisins
Biocatalysts
Surface-Active Agents
Organic solvents
Peptides
Surface active agents
Peptide Hydrolases
Enzymes
Amides
Reaction rates
Amino acids
Hydrolysis
Catalyst activity
Alcohols
Water
Nonionic surfactants
Dimethylformamide
Esters
Solubility
Amino Acids

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology

Cite this

Surfactant-protease complex as a novel biocatalyst for peptide synthesis in hydrophilic organic solvents. / Okazaki, Shin ya; Goto, Masahiro; Furusaki, Shintaro.

In: Enzyme and Microbial Technology, Vol. 26, No. 2-4, 01.01.2000, p. 159-164.

Research output: Contribution to journalArticle

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