Kinetics of the hydrolysis of benzoylglycine ethyl ester catalyzed by papain

L.A.A.E. Sluyterman

Research output: Contribution to journalArticleAcademic

27 Citations (Scopus)

Abstract

The hydrolysis of benzoylglycine ethyl ester has been investigated. The kinetics can be described by the conventional Michaelis-Menten scheme. The substrate appeared to be only four times less sensitive than the best known substrate benzoylarginine ethyl ester. The pH dependence of the hydrolysis of benzoylglycine ethyl ester closely resembled equivalent data for other papain (EC 3.4.4.10) substrates as reported in the literature. The binding of the substrate was maximal at pH 6, half-maximal at pH 4 (ascribed to a carboxyl group in the active centre) and at pH 8 (ascribed to a sulfhydryl group or an imidazolium group). The vmax was constant in the range pH 4.2–8.4 , which indicates that histidine is not involved in the catalytic step as it is with trypsin (EC 3.4.4.4) and chymotrypsin (EC 3.4.4.5). The experimental procedure is described in detail. The purity of the papain was checked by chromatography on a CM-cellulose column, and by electrophoresis in starch gel.
Original languageEnglish
Pages (from-to)305-315
Number of pages11
JournalBiochimica et Biophysica Acta, Specialized Section on Enzymological Subjects
Volume85
Issue number2
DOIs
Publication statusPublished - 1964

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Papain
Hydrolysis
Esters
Kinetics
Substrates
Chymotrypsin
Chromatography
Electrophoresis
Histidine
Cellulose
Starch
Trypsin
Gels
hippuric acid

Cite this

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title = "Kinetics of the hydrolysis of benzoylglycine ethyl ester catalyzed by papain",
abstract = "The hydrolysis of benzoylglycine ethyl ester has been investigated. The kinetics can be described by the conventional Michaelis-Menten scheme. The substrate appeared to be only four times less sensitive than the best known substrate benzoylarginine ethyl ester. The pH dependence of the hydrolysis of benzoylglycine ethyl ester closely resembled equivalent data for other papain (EC 3.4.4.10) substrates as reported in the literature. The binding of the substrate was maximal at pH 6, half-maximal at pH 4 (ascribed to a carboxyl group in the active centre) and at pH 8 (ascribed to a sulfhydryl group or an imidazolium group). The vmax was constant in the range pH 4.2–8.4 , which indicates that histidine is not involved in the catalytic step as it is with trypsin (EC 3.4.4.4) and chymotrypsin (EC 3.4.4.5). The experimental procedure is described in detail. The purity of the papain was checked by chromatography on a CM-cellulose column, and by electrophoresis in starch gel.",
author = "L.A.A.E. Sluyterman",
year = "1964",
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journal = "Biochimica et Biophysica Acta, Specialized Section on Enzymological Subjects",
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Kinetics of the hydrolysis of benzoylglycine ethyl ester catalyzed by papain. / Sluyterman, L.A.A.E.

In: Biochimica et Biophysica Acta, Specialized Section on Enzymological Subjects, Vol. 85, No. 2, 1964, p. 305-315.

Research output: Contribution to journalArticleAcademic

TY - JOUR

T1 - Kinetics of the hydrolysis of benzoylglycine ethyl ester catalyzed by papain

AU - Sluyterman, L.A.A.E.

PY - 1964

Y1 - 1964

N2 - The hydrolysis of benzoylglycine ethyl ester has been investigated. The kinetics can be described by the conventional Michaelis-Menten scheme. The substrate appeared to be only four times less sensitive than the best known substrate benzoylarginine ethyl ester. The pH dependence of the hydrolysis of benzoylglycine ethyl ester closely resembled equivalent data for other papain (EC 3.4.4.10) substrates as reported in the literature. The binding of the substrate was maximal at pH 6, half-maximal at pH 4 (ascribed to a carboxyl group in the active centre) and at pH 8 (ascribed to a sulfhydryl group or an imidazolium group). The vmax was constant in the range pH 4.2–8.4 , which indicates that histidine is not involved in the catalytic step as it is with trypsin (EC 3.4.4.4) and chymotrypsin (EC 3.4.4.5). The experimental procedure is described in detail. The purity of the papain was checked by chromatography on a CM-cellulose column, and by electrophoresis in starch gel.

AB - The hydrolysis of benzoylglycine ethyl ester has been investigated. The kinetics can be described by the conventional Michaelis-Menten scheme. The substrate appeared to be only four times less sensitive than the best known substrate benzoylarginine ethyl ester. The pH dependence of the hydrolysis of benzoylglycine ethyl ester closely resembled equivalent data for other papain (EC 3.4.4.10) substrates as reported in the literature. The binding of the substrate was maximal at pH 6, half-maximal at pH 4 (ascribed to a carboxyl group in the active centre) and at pH 8 (ascribed to a sulfhydryl group or an imidazolium group). The vmax was constant in the range pH 4.2–8.4 , which indicates that histidine is not involved in the catalytic step as it is with trypsin (EC 3.4.4.4) and chymotrypsin (EC 3.4.4.5). The experimental procedure is described in detail. The purity of the papain was checked by chromatography on a CM-cellulose column, and by electrophoresis in starch gel.

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DO - 10.1016/0926-6569(64)90251-2

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JO - Biochimica et Biophysica Acta, Specialized Section on Enzymological Subjects

JF - Biochimica et Biophysica Acta, Specialized Section on Enzymological Subjects

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