A sensitive HPLC method for measuring bacterial proteolysis and proteinase activity in UHT milk

A sensitive quantitative reversed-phase HPLC method is described for measuring bacterial proteolysis and proteinase activity in UHT milk. The analysis is performed on a TCA filtrate of the milk. The optimum concentration of TCA was found to be 4%; at lower concentrations, non-precipitated protein bl...

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Published inFood research international Vol. 39; no. 7; pp. 823 - 830
Main Authors Le, T.X., Datta, N., Deeth, H.C.
Format Journal Article
LanguageEnglish
Published Oxford Elsevier Ltd 01.08.2006
Elsevier
Subjects
Bacteria
bacterial contamination
Bacterial proteinase
Biological and medical sciences
enzyme activity
Fluorescamine
food contamination
Food industries
Fundamental and applied biological sciences. Psychology
HPLC
Milk and cheese industries. Ice creams
peptides
Proteinase activity
proteinases
Proteolysis
psychrotrophic bacteria
reversed-phase high performance liquid chromatography
trichloroacetic acid
UHT milk
Fluorescamine
Proteolysis
Proteinase activity
Bacterial proteinase
HPLC
Peptidases
Enzyme
Dairy product
HPLC chromatography
Hydrolases
UHT milk
Method
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Abstract A sensitive quantitative reversed-phase HPLC method is described for measuring bacterial proteolysis and proteinase activity in UHT milk. The analysis is performed on a TCA filtrate of the milk. The optimum concentration of TCA was found to be 4%; at lower concentrations, non-precipitated protein blocked the HPLC while higher concentrations yielded lower amounts of peptides. The method showed greater sensitivity and reproducibility than a fluorescamine-based method. Quantification of the HPLC method was achieved by use of an external dipeptide standard or a standard proteinase.
AbstractList A sensitive quantitative reversed-phase HPLC method is described for measuring bacterial proteolysis and proteinase activity in UHT milk. The analysis is performed on a TCA filtrate of the milk. The optimum concentration of TCA was found to be 4%; at lower concentrations, non-precipitated protein blocked the HPLC while higher concentrations yielded lower amounts of peptides. The method showed greater sensitivity and reproducibility than a fluorescamine- based method. Quantification of the HPLC method was achieved by use of an external dipeptide standard or a standard proteinase.
Author Le, T.X.
Datta, N.
Deeth, H.C.
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Cites_doi 10.1017/S0022029901005301
10.1205/096030801753252261
10.1111/j.1365-2621.1979.tb08487.x
10.1016/S0958-6946(97)00072-1
10.3168/jds.S0022-0302(67)87636-7
10.1023/A:1026357009886
10.1016/S0023-6438(02)00214-1
10.1111/j.1399-3011.1989.tb00227.x
10.1017/S0022029900027394
10.3168/jds.S0022-0302(79)83499-2
10.3168/jds.S0022-0302(75)84645-5
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Issue 7
Keywords Fluorescamine
Proteolysis
Proteinase activity
Bacterial proteinase
HPLC
Peptidases
Enzyme
Dairy product
HPLC chromatography
Hydrolases
UHT milk
Method
Language English
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Haryani, Datta, Elliott, Deeth (bib9) 2003; 58
Kocak, Zadow (bib10) 1985; 40
López-Fandiño, Olano, San Jose, Ramos (bib12) 1993; 60
Datta, Deeth (bib5) 2003; 36
Pearce (bib14) 1979; 14
Beeby (bib2) 1980; 15
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Datta, Deeth (bib4) 2001; 79
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Lieske, Konrad, Faber (bib11) 1997; 7
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López-Fandiño (10.1016/j.foodres.2006.03.008_bib12) 1993; 60
Sivaraman (10.1016/j.foodres.2006.03.008_bib15) 1997; 16
Yvon (10.1016/j.foodres.2006.03.008_bib16) 1989; 34
Mitchell (10.1016/j.foodres.2006.03.008_bib13) 1985; 20
Kocak (10.1016/j.foodres.2006.03.008_bib10) 1985; 40
Datta (10.1016/j.foodres.2006.03.008_bib5) 2003; 36
Greenberg (10.1016/j.foodres.2006.03.008_bib8) 1979; 44
Deeth (10.1016/j.foodres.2006.03.008_bib6) 2002; 69
Haryani (10.1016/j.foodres.2006.03.008_bib9) 2003; 58
Pearce (10.1016/j.foodres.2006.03.008_bib14) 1979; 14
Fox (10.1016/j.foodres.2006.03.008_bib7) 1967; 50
Beeby (10.1016/j.foodres.2006.03.008_bib2) 1980; 15
Chism (10.1016/j.foodres.2006.03.008_bib3) 1979; 62
Lieske (10.1016/j.foodres.2006.03.008_bib11) 1997; 7
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Snippet A sensitive quantitative reversed-phase HPLC method is described for measuring bacterial proteolysis and proteinase activity in UHT milk. The analysis is...
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SubjectTerms Bacteria
bacterial contamination
Bacterial proteinase
Biological and medical sciences
enzyme activity
Fluorescamine
food contamination
Food industries
Fundamental and applied biological sciences. Psychology
HPLC
Milk and cheese industries. Ice creams
peptides
Proteinase activity
proteinases
Proteolysis
psychrotrophic bacteria
reversed-phase high performance liquid chromatography
trichloroacetic acid
UHT milk
Title A sensitive HPLC method for measuring bacterial proteolysis and proteinase activity in UHT milk
URI https://dx.doi.org/10.1016/j.foodres.2006.03.008
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