Purification and characterization of chymotrypsin-like enzyme from rat plasma

R. Senthil Kumar, T. N. Pattabiraman

Research output: Contribution to journalArticle

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Abstract

A chymotrypsin-like enzyme was purified from rat plasma, involving ammonium sulfate fractionation and chromatographgy on CM-sephadex and red sepharose. The purified enzyme effectively hydrolysed the ester substrates for chymotrypsin (N-acetyl L-tyrosine ethyl ester and N-acetyl L-tryptophan ethyl ester). The Km values for the two substrates were 2.2×10-3M and 9.0×10-3M respectively. The hydrolytic activity of the enzyme was inhibited by phenylmethyl sulfonyl fluoride and tosylphenylalanine chloromethylketone, suggesting the presence of serine and histidine at the active centre. The enzyme exhibited anionic nature and possessed a high molecular weight (MW 71,000) as observed by gel exclusion chromatography on Sephadex G-200. The enzyme was stable upon exposure to pH 7.0-9.0, but was inactivated upon heat treatment at 60°C for 5 min.

Original languageEnglish
Pages (from-to)152-157
Number of pages6
JournalIndian Journal of Clinical Biochemistry
Volume11
Issue number2
DOIs
Publication statusPublished - 01-07-1996
Externally publishedYes

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Purification
Rats
Plasmas
Enzymes
Gel Chromatography
Chymotrypsin
Ammonium Sulfate
Substrates
Fractionation
Chromatography
Histidine
Tryptophan
Sepharose
Serine
Esters
Hot Temperature
Molecular Weight
Gels
Molecular weight
Heat treatment

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry

Cite this

Kumar, R. Senthil ; Pattabiraman, T. N. / Purification and characterization of chymotrypsin-like enzyme from rat plasma. In: Indian Journal of Clinical Biochemistry. 1996 ; Vol. 11, No. 2. pp. 152-157.
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Purification and characterization of chymotrypsin-like enzyme from rat plasma. / Kumar, R. Senthil; Pattabiraman, T. N.

In: Indian Journal of Clinical Biochemistry, Vol. 11, No. 2, 01.07.1996, p. 152-157.

Research output: Contribution to journalArticle

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