Properties of elastase from Atlantic cod a cold-adapted proteinase

Bjarni Ásgeirsson; Jón Bragi Bjarnason

doi:10.1016/0167-4838(93)90116-9

Properties of elastase from Atlantic cod a cold-adapted proteinase

Bjarni Ásgeirsson
, Jón Bragi Bjarnason

Faculty of Physical Sciences

University of Iceland

Research output: Contribution to journal › Article › peer-review

Abstract

An intestinal elastase was purified from Atlantic cod (Gadus morhua) of apparent molecular mass 24.8 kDa as determined by SDS-PAGE and with isoelectric point above pI 9.3. Heat stability and stability towards acidic pH was reduced in the cod enzyme as compared with porcine intestinal elastase. N-terminal amino-acid sequence analysis of cod elastase showed considerable similarity with porcine elastase. The cod enzyme was less sensitive to phenylmethylsulfonyl fluoride inhibition than porcine elastase, but sensitivity towards other inhibitors was similar. Kinetic properties were examined using the substrate Suc-Ala-Ala-Ala-p-nitroanilide and the cod enzyme was found to have more than 2-times higher turnover rate (k_cat) as compared with the porcine enzyme, and slightly higher Km values. Thus, the catalytic efficiency ( k_cat K_m) of Atlantic cod elastase was about 2-times higher than observed with porcine elastase, which indicates an adaptive response towards the low temperature environmental in which the cod lives. Substrate specificity was studied by digestion of oxidized B-chain of insulin and by using synthetic substrates. Digestion was most rapid at the carbonyl side of alanine residues, but also occurred at valine and leucine residues.

Original language	English
Pages (from-to)	91-100
Number of pages	10
Journal	Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
Volume	1164
Issue number	1
DOIs	https://doi.org/10.1016/0167-4838(93)90116-9
Publication status	Published - 24 Jun 1993

Bibliographical note

Funding Information: The authors wish to thank Gunnhildur Gislad6ttir for performing specificity experiments with insulin B-chain, and Dr. Jay W. Fox for the amino-acid analysis and N-terminal sequence determination. This work was supported by The National Research Council of Iceland and Nordisk Industrifond.

Other keywords

(G. morhua)
Cold adaptation
Elastase
Poikilotherm
Serine proteinase

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10.1016/0167-4838(93)90116-9

Cite this

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title = "Properties of elastase from Atlantic cod a cold-adapted proteinase",

abstract = "An intestinal elastase was purified from Atlantic cod (Gadus morhua) of apparent molecular mass 24.8 kDa as determined by SDS-PAGE and with isoelectric point above pI 9.3. Heat stability and stability towards acidic pH was reduced in the cod enzyme as compared with porcine intestinal elastase. N-terminal amino-acid sequence analysis of cod elastase showed considerable similarity with porcine elastase. The cod enzyme was less sensitive to phenylmethylsulfonyl fluoride inhibition than porcine elastase, but sensitivity towards other inhibitors was similar. Kinetic properties were examined using the substrate Suc-Ala-Ala-Ala-p-nitroanilide and the cod enzyme was found to have more than 2-times higher turnover rate (kcat) as compared with the porcine enzyme, and slightly higher Km values. Thus, the catalytic efficiency ( kcat Km) of Atlantic cod elastase was about 2-times higher than observed with porcine elastase, which indicates an adaptive response towards the low temperature environmental in which the cod lives. Substrate specificity was studied by digestion of oxidized B-chain of insulin and by using synthetic substrates. Digestion was most rapid at the carbonyl side of alanine residues, but also occurred at valine and leucine residues.",

keywords = "(G. morhua), Cold adaptation, Elastase, Poikilotherm, Serine proteinase",

author = "Bjarni {\'A}sgeirsson and Bjarnason, \{J{\'o}n Bragi\}",

note = "Funding Information: The authors wish to thank Gunnhildur Gislad6ttir for performing specificity experiments with insulin B-chain, and Dr. Jay W. Fox for the amino-acid analysis and N-terminal sequence determination. This work was supported by The National Research Council of Iceland and Nordisk Industrifond.",

year = "1993",

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doi = "10.1016/0167-4838(93)90116-9",

language = "English",

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pages = "91--100",

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AU - Ásgeirsson, Bjarni

AU - Bjarnason, Jón Bragi

N1 - Funding Information: The authors wish to thank Gunnhildur Gislad6ttir for performing specificity experiments with insulin B-chain, and Dr. Jay W. Fox for the amino-acid analysis and N-terminal sequence determination. This work was supported by The National Research Council of Iceland and Nordisk Industrifond.

PY - 1993/6/24

Y1 - 1993/6/24

N2 - An intestinal elastase was purified from Atlantic cod (Gadus morhua) of apparent molecular mass 24.8 kDa as determined by SDS-PAGE and with isoelectric point above pI 9.3. Heat stability and stability towards acidic pH was reduced in the cod enzyme as compared with porcine intestinal elastase. N-terminal amino-acid sequence analysis of cod elastase showed considerable similarity with porcine elastase. The cod enzyme was less sensitive to phenylmethylsulfonyl fluoride inhibition than porcine elastase, but sensitivity towards other inhibitors was similar. Kinetic properties were examined using the substrate Suc-Ala-Ala-Ala-p-nitroanilide and the cod enzyme was found to have more than 2-times higher turnover rate (kcat) as compared with the porcine enzyme, and slightly higher Km values. Thus, the catalytic efficiency ( kcat Km) of Atlantic cod elastase was about 2-times higher than observed with porcine elastase, which indicates an adaptive response towards the low temperature environmental in which the cod lives. Substrate specificity was studied by digestion of oxidized B-chain of insulin and by using synthetic substrates. Digestion was most rapid at the carbonyl side of alanine residues, but also occurred at valine and leucine residues.

AB - An intestinal elastase was purified from Atlantic cod (Gadus morhua) of apparent molecular mass 24.8 kDa as determined by SDS-PAGE and with isoelectric point above pI 9.3. Heat stability and stability towards acidic pH was reduced in the cod enzyme as compared with porcine intestinal elastase. N-terminal amino-acid sequence analysis of cod elastase showed considerable similarity with porcine elastase. The cod enzyme was less sensitive to phenylmethylsulfonyl fluoride inhibition than porcine elastase, but sensitivity towards other inhibitors was similar. Kinetic properties were examined using the substrate Suc-Ala-Ala-Ala-p-nitroanilide and the cod enzyme was found to have more than 2-times higher turnover rate (kcat) as compared with the porcine enzyme, and slightly higher Km values. Thus, the catalytic efficiency ( kcat Km) of Atlantic cod elastase was about 2-times higher than observed with porcine elastase, which indicates an adaptive response towards the low temperature environmental in which the cod lives. Substrate specificity was studied by digestion of oxidized B-chain of insulin and by using synthetic substrates. Digestion was most rapid at the carbonyl side of alanine residues, but also occurred at valine and leucine residues.

KW - (G. morhua)

KW - Cold adaptation

KW - Elastase

KW - Poikilotherm

KW - Serine proteinase

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JF - Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular

IS - 1

ER -

Properties of elastase from Atlantic cod a cold-adapted proteinase

Abstract

Bibliographical note

Other keywords

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