N-(Benzyloxycarbonyl)-l-aspartyl-l-phenylalanine methyl ester production by immobilized Nocardia opaca: Thermolysin under hydrogen high pressure in a water-organic solvent system

Noriyuki Nakamura, Anny Sulaswatty, Shigeo Nishimura, Takanori Kitamura, Masao Tsuji, Toshio Kawaguchi, Tadashi Matsunaga

Research output: Contribution to journalArticlepeer-review

11 Citations (Scopus)

Abstract

N-(Benzyloxycarbonyl)-l-aspartyl-l-phenylalanine methyl ester (Z-APM) has been produced from phenylpyruvate methyl ester (PPyOMe) and N-(benzyloxycarbonyl)-aspartic acid (Z-Asp) under hydrogen high pressure in a water-ethyl acetate system. Z-APM was produced in a two step reaction using immobilized Nocardia opaca and thermolysin. Phenylalanine methyl ester (PheOMe) was produced from PPyOMe, NH4Cl and H2 by immobilized N. opaca. The PheOMe production rate increased linearly with increasing hydrogen pressure. The PheOMe production rate was 0.41 μmol/min/g dry cells under 100 atm of hydrogen. Z-APM was produced from PheOMe and Z-Asp by immobilized thermolysin. The highest activity (22.2 μmol/min/g thermolysin) was obtained by coupling N. opaca immobilized in calcium alginate and thermolysin immobilized in photo-crosslinkable resin prepolymer ENTP-4000. When repeated use was carried out without reactivation of the immobilized N. opaca, Z-APM production was not observed after triple use. Therefore, immobilized N. opaca was separated from immobilized thermolysin and reactivated periodically in nutrient broth for 20 h. As a result, the immobilized system retained 72% of the initial activity after being used 5 times for over 100 h.

Original languageEnglish
Pages (from-to)399-403
Number of pages5
JournalJournal of Fermentation and Bioengineering
Volume67
Issue number6
DOIs
Publication statusPublished - 1989
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

Fingerprint Dive into the research topics of 'N-(Benzyloxycarbonyl)-l-aspartyl-l-phenylalanine methyl ester production by immobilized Nocardia opaca: Thermolysin under hydrogen high pressure in a water-organic solvent system'. Together they form a unique fingerprint.

Cite this