Chemical Modification of Earthworm Fibrinolytic Enzyme with Human Serum Albumin Fragment and Characterization of the Protease as a Therapeutic Enzyme

Nobuyoshi Nakajima, Kohji Ishihara, Manabu Sugimoto, Hiroyuki Sumi, Katsuhiko Mikuni, Hiroki Hamada

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

The strongest fibrinolytic protease (F-III-2) in the six enzyme proteins purified from earthworm, Lumbricus rubellus [N. Nakajima et al., Biosci. Biotech. Biochem., 57, 1726-1730 (1993)] has been modified chemically with fragmented human serum albumin (mol. wt., 10,000-30,000). The modified enzyme lost the antigenicity of the native enzyme and reacted with the antisera against human serum albumin, the human serum albumin fragments, and the conjugate with the native enzyme to form precipitation lines, which fused with each other. The conjugate was significantly more resistant to inactivation by protease inhibitors in rat plasma. The enzyme was a non-hemorrhagic protein and did not induce platelet aggregation. The enzyme kept potent proteolytic activity for fibrin and fibrinogen than that of human plasmin. The enzyme easily solubilized actual fibrin clots (thrombi) of whole blood induced by thrombin in a rat's vena cava. The continuous fibrinolysis for fibrin suspension in an enzyme reactor system using the modified enzyme immobilized to oxirane-activated acrylic beads has been achieved without any inactivation of the activity at least for more than 1 month. The N-terminal amino acid sequence of the protein was also investigated and the sequence showed local similarity to those of the serine proteases such as plasmin and chymotrypsin.

Original languageEnglish
Pages (from-to)293-300
Number of pages8
JournalBioscience, Biotechnology and Biochemistry
Volume60
Issue number2
Publication statusPublished - Feb 1996

Fingerprint

Oligochaeta
Chemical modification
earthworms
Serum Albumin
Peptide Hydrolases
proteinases
therapeutics
Enzymes
enzymes
fibrin
Fibrinolysin
Fibrin
plasmin
pitrilysin
Therapeutics
Rats
inactivation
Lumbricus rubellus
fibrinolysis
ethylene oxide

Keywords

  • Chemical modification
  • Fibrinolytic enzyme (of earthworm)
  • Immobilized enzyme
  • Protein sequence
  • Serine protease (of earthworm)

ASJC Scopus subject areas

  • Food Science
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biotechnology
  • Chemistry (miscellaneous)
  • Applied Microbiology and Biotechnology

Cite this

Chemical Modification of Earthworm Fibrinolytic Enzyme with Human Serum Albumin Fragment and Characterization of the Protease as a Therapeutic Enzyme. / Nakajima, Nobuyoshi; Ishihara, Kohji; Sugimoto, Manabu; Sumi, Hiroyuki; Mikuni, Katsuhiko; Hamada, Hiroki.

In: Bioscience, Biotechnology and Biochemistry, Vol. 60, No. 2, 02.1996, p. 293-300.

Research output: Contribution to journalArticle

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