Improved stability and catalytic activity of chemically modified papain in aqueous organic solvents

András Szabó, M. Kotormán, I. Laczkó, L. Mária Simon

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Papain was modified with the anhydrides of various monocarboxylic (acetic or propionic) and dicarboxylic (citraconic, maleic or succinic) acids. 7-10 of the 11 primary amino groups of the enzyme were modified. The organic solvent tolerances of the modified enzyme forms were increased (especially in the concentration range of 10-60%) in comparison with the unmodified enzyme. Acylation enhanced the catalytic activity and stability of papain both in buffer and in aqueous organic solvents (ethanol and acetonitrile). Decrease of the positive charges on the surface of papain resulted in a higher enzyme stability than when they were replaced by negative charges. The kinetic parameters revealed that in aqueous ethanol the maximum rates (Vmax) and Michaelis constants (KM) of the modified papain forms were increased, and higher catalytic efficiencies (kcat/KM) were detected as compared with the native enzyme. The results of near-UV circular dichroism and tryptophan fluorescence spectroscopic studies suggested that the modifications caused only local changes around the aromatic residues. The modified enzyme forms led to higher N-acetyl-l-tyrosine ethyl ester synthesis conversions in aqueous ethanol; acetyl and propionyl papain furnishing the highest productivity.

Original languageEnglish
Pages (from-to)199-204
Number of pages6
JournalProcess Biochemistry
Volume44
Issue number2
DOIs
Publication statusPublished - Feb 2009

Fingerprint

Papain
Organic solvents
Catalyst activity
Enzymes
Ethanol
Succinates
Enzyme Stability
Acylation
Anhydrides
Circular Dichroism
Tryptophan
Acetic Acid
Dichroism
Acetonitrile
Buffers
Kinetic parameters
Fluorescence
Esters
Productivity
Acids

Keywords

  • Amino acid ester synthesis
  • CD and fluorescence spectroscopy
  • Chemical modification
  • Organic acid anhydrides
  • Organic solvents
  • Stability of papain

ASJC Scopus subject areas

  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Bioengineering

Cite this

Improved stability and catalytic activity of chemically modified papain in aqueous organic solvents. / Szabó, András; Kotormán, M.; Laczkó, I.; Simon, L. Mária.

In: Process Biochemistry, Vol. 44, No. 2, 02.2009, p. 199-204.

Research output: Contribution to journalArticle

@article{39c4bcb3f5484769af6c6c72dd0b62e5,
title = "Improved stability and catalytic activity of chemically modified papain in aqueous organic solvents",
abstract = "Papain was modified with the anhydrides of various monocarboxylic (acetic or propionic) and dicarboxylic (citraconic, maleic or succinic) acids. 7-10 of the 11 primary amino groups of the enzyme were modified. The organic solvent tolerances of the modified enzyme forms were increased (especially in the concentration range of 10-60{\%}) in comparison with the unmodified enzyme. Acylation enhanced the catalytic activity and stability of papain both in buffer and in aqueous organic solvents (ethanol and acetonitrile). Decrease of the positive charges on the surface of papain resulted in a higher enzyme stability than when they were replaced by negative charges. The kinetic parameters revealed that in aqueous ethanol the maximum rates (Vmax) and Michaelis constants (KM) of the modified papain forms were increased, and higher catalytic efficiencies (kcat/KM) were detected as compared with the native enzyme. The results of near-UV circular dichroism and tryptophan fluorescence spectroscopic studies suggested that the modifications caused only local changes around the aromatic residues. The modified enzyme forms led to higher N-acetyl-l-tyrosine ethyl ester synthesis conversions in aqueous ethanol; acetyl and propionyl papain furnishing the highest productivity.",
keywords = "Amino acid ester synthesis, CD and fluorescence spectroscopy, Chemical modification, Organic acid anhydrides, Organic solvents, Stability of papain",
author = "Andr{\'a}s Szab{\'o} and M. Kotorm{\'a}n and I. Laczk{\'o} and Simon, {L. M{\'a}ria}",
year = "2009",
month = "2",
doi = "10.1016/j.procbio.2008.10.008",
language = "English",
volume = "44",
pages = "199--204",
journal = "Process Biochemistry",
issn = "1359-5113",
publisher = "Elsevier BV",
number = "2",

}

TY - JOUR

T1 - Improved stability and catalytic activity of chemically modified papain in aqueous organic solvents

AU - Szabó, András

AU - Kotormán, M.

AU - Laczkó, I.

AU - Simon, L. Mária

PY - 2009/2

Y1 - 2009/2

N2 - Papain was modified with the anhydrides of various monocarboxylic (acetic or propionic) and dicarboxylic (citraconic, maleic or succinic) acids. 7-10 of the 11 primary amino groups of the enzyme were modified. The organic solvent tolerances of the modified enzyme forms were increased (especially in the concentration range of 10-60%) in comparison with the unmodified enzyme. Acylation enhanced the catalytic activity and stability of papain both in buffer and in aqueous organic solvents (ethanol and acetonitrile). Decrease of the positive charges on the surface of papain resulted in a higher enzyme stability than when they were replaced by negative charges. The kinetic parameters revealed that in aqueous ethanol the maximum rates (Vmax) and Michaelis constants (KM) of the modified papain forms were increased, and higher catalytic efficiencies (kcat/KM) were detected as compared with the native enzyme. The results of near-UV circular dichroism and tryptophan fluorescence spectroscopic studies suggested that the modifications caused only local changes around the aromatic residues. The modified enzyme forms led to higher N-acetyl-l-tyrosine ethyl ester synthesis conversions in aqueous ethanol; acetyl and propionyl papain furnishing the highest productivity.

AB - Papain was modified with the anhydrides of various monocarboxylic (acetic or propionic) and dicarboxylic (citraconic, maleic or succinic) acids. 7-10 of the 11 primary amino groups of the enzyme were modified. The organic solvent tolerances of the modified enzyme forms were increased (especially in the concentration range of 10-60%) in comparison with the unmodified enzyme. Acylation enhanced the catalytic activity and stability of papain both in buffer and in aqueous organic solvents (ethanol and acetonitrile). Decrease of the positive charges on the surface of papain resulted in a higher enzyme stability than when they were replaced by negative charges. The kinetic parameters revealed that in aqueous ethanol the maximum rates (Vmax) and Michaelis constants (KM) of the modified papain forms were increased, and higher catalytic efficiencies (kcat/KM) were detected as compared with the native enzyme. The results of near-UV circular dichroism and tryptophan fluorescence spectroscopic studies suggested that the modifications caused only local changes around the aromatic residues. The modified enzyme forms led to higher N-acetyl-l-tyrosine ethyl ester synthesis conversions in aqueous ethanol; acetyl and propionyl papain furnishing the highest productivity.

KW - Amino acid ester synthesis

KW - CD and fluorescence spectroscopy

KW - Chemical modification

KW - Organic acid anhydrides

KW - Organic solvents

KW - Stability of papain

UR - http://www.scopus.com/inward/record.url?scp=58149260294&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=58149260294&partnerID=8YFLogxK

U2 - 10.1016/j.procbio.2008.10.008

DO - 10.1016/j.procbio.2008.10.008

M3 - Article

AN - SCOPUS:58149260294

VL - 44

SP - 199

EP - 204

JO - Process Biochemistry

JF - Process Biochemistry

SN - 1359-5113

IS - 2

ER -