Isolation, purification and characterization of a trypsin from the pyloric ceca of mullet (Mugil cephalus)

N. Guizani, R. S. Rolle, M. R. Marshall, C. I. Wei

Research output: Contribution to journalArticle

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Abstract

1. 1. A protease classified as trypsin on the basis of its mol. wt of 24,000, its ability to hydrolyze synthetic substrates N-α-benzoyl-arginine-p-nitroanilide (BAPA) and tosylarginine methyl ester (TAME), and the inhibitory effects of methyl sulfonyl fluoride (PMSF), SBTI, aprotinin and benzamidine on its activity was isolated from the pyloric ceca of mullet. 2. 2. The enzyme exhibited optimal activity at pH 8 and a temperature of 55°C and was stable in the pH range of 7.5-9.0. The Arrhenius energy of activation (Ea) for BAPA hydrolysis by this enzyme was 7.4 kcal/mol. 3. 3. The enzyme showed greater affinity for TAME (K′m = 0.19 mM) as a substrate than for BAPA (K′m = 0.49 mM), and preferential hydrolysis of ester bonds (Vmax = 2640 TAME units/μmmol enzyme) than amide bonds (Vmax = 400 BAPA units/μmol enzyme).

Original languageEnglish
Pages (from-to)517-521
Number of pages5
JournalComparative Biochemistry and Physiology - B Biochemistry and Molecular Biology
Volume98
Issue number4
DOIs
Publication statusPublished - 1991

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Smegmamorpha
Tosylarginine Methyl Ester
Cecum
Trypsin
Purification
Enzymes
Hydrolysis
Aprotinin
Substrates
Amides
Arginine
Esters
Peptide Hydrolases
Chemical activation
Temperature

ASJC Scopus subject areas

  • Biochemistry
  • Physiology

Cite this

Isolation, purification and characterization of a trypsin from the pyloric ceca of mullet (Mugil cephalus). / Guizani, N.; Rolle, R. S.; Marshall, M. R.; Wei, C. I.

In: Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology, Vol. 98, No. 4, 1991, p. 517-521.

Research output: Contribution to journalArticle

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AU - Wei, C. I.

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