Hostname: page-component-78c5997874-j824f Total loading time: 0 Render date: 2024-11-17T19:37:22.598Z Has data issue: false hasContentIssue false
  • English
  • Français

Isolation and characterization of heat stable proteinases from Pseudomonas isolate AFT 21

Published online by Cambridge University Press:  01 June 2009

Leszek Stepaniak  and
Patrick F. Fox
Leszek Stepaniak
Affiliation:
Department of Dairy and Food Chemistry, University College, Cork, Irish Republic
Patrick F. Fox
Affiliation:
Department of Dairy and Food Chemistry, University College, Cork, Irish Republic
Get access Rights & Permissions [Opens in a new window]

Summary

Pseudomonas strain AFT 21 produced three heat stable extracellular proteinases in milk and nutrient broth at 7 or 21 °C, but the proportions depended on medium and cultivation temperature. The three proteinases were EDTA- and o-phenanthroline-sensitive metalloenzymes and were not inhibited by N-ethylmaleimide or phosphoramidon. Proteinases I and II showed maximum activity at pH 7–7·5 and proteinase III at pH 8·5. All three enzymes showed maximum activity at 45–47·5 °C, but had relatively high (19–27% of maximum) activity at 4 °C. They were unstable at 55 °C in phosphate buffer, pH 6·6, or synthetic milk ultrafiltrate (SMUF) containing 12 mmol Ca2+, but were stabilized by short preheating at 100 °C. They were extremely heat stable in both phosphate buffer and SMUF, pH 6·6, at 70–150 °C. Their D-values at 140 °C were 69, 54 and 80 s respectively. The Z-values for Pseudomonas AFT 21 proteinase III in phosphate buffer and SMUF were 29·7 and 30·3 °C respectively; the corresponding activation energies for inactivation were 8·7 × 104 J mol-1 and 9·2 x 104 J mol-1.

Type
Original Articles
Information
Journal of Dairy Research , Volume 52 , Issue 1 , February 1985 , pp. 77 - 89
Copyright
Copyright © Proprietors of Journal of Dairy Research 1985

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Adams, D. M., Barach, J. T. & Speck, M. L. 1975 Heat resistant proteases produced in milk by psychrotrophic bacteria of dairy origin. Journal of Dairy Science 58 828–834Google Scholar
Alichanidis, E. & Andrews, A. T. 1977 Some properties of the extracellular protease produced by the psychrotrophic bacterium Pseudomonas fluorescens strain AR-11. Biochimica et Biophysica Acta 485 424–433Google Scholar
Barach, J. T. & Adams, D. M. 1977 Thermostability at ultrahigh temperatures of thermolysin and a protease from a psychrotrophic Pseudomonas. Biochimica et Biophysica Acta 485 417–423Google Scholar
Barach, J. T., Adams, D. M. & Speck, M. L. 1976 a Stabilization of a psychrotrophic Pseudomonas protease by calcium against thermal inactivation in milk at ultrahigh temperature. Applied and Environmental Microbiology 31 875–879Google Scholar
Barach, J. T., Adams, D. M. & Speck, M. L. 1976 b Low temperature inactivation in milk of heat-resistant proteases from psychrotrophic bacteria. Journal of Dairy Science 59 391–395Google Scholar
Barach, J. T., Adams, D. M. & Speck, M. L. 1978 Mechanism of low temperature inactivation of a heat-resistant bacterial protease in milk. Journal of Dairy Science 61 523–528Google Scholar
Cogan, T. M. 1977 A review of heat resistant lipases and proteinases and the quality of dairy products. Irish Journal of Food Science and Technology 1 95–105Google Scholar
Cousin, M. A. 1982 Presence and activity of psychrotrophic microorganisms in milk and dairy products: A review. Journal of Food Protection 45 172–207Google Scholar
Dawson, R. M. C., Elliott, D. C., Elliott, W. H. & Jones, K. M. 1969 Data for Biochemical Research, 2nd edn. p. 485. Oxford: Clarendon PressGoogle Scholar
Fields, R. 1972 The rapid determination of amino groups with TNBS. Methods in Enzymology 25 464–468Google Scholar
Fox, P. F. 1981 Proteinases in dairy technology. Netherlands Milk and Dairy Journal 35 233–253Google Scholar
Gebre-Egziabher, A., Humbert, E. S. & Blanknagel, G. 1980 Heat-stable proteases from psychrotrophs in milk. Journal of Food Protection 43 197–200Google Scholar
Griffiths, M. W., Phillips, J. D. & Muir, D. D. 1981 Thermostability of proteases and lipases from a number of species of psychrotrophic bacteria of dairy origin. Journal of Applied Bacteriology 50 289–303Google Scholar
Gripon, J. C., Auberger, B. & Lenoir, J. 1980 Metalloproteases from Penicillium caseicolum and P. roqueforti: comparison of specificity and chemical characterization. International Journal of Biochemistry 12 451–455Google Scholar
Jenness, R. & Koops, J. 1962 Preparation and properties of a salt solution which simulates milk ultrafiltrate. Netherlands Milk and Dairy Journal 16 153–164Google Scholar
Komiyama, T., Aoyagi, T., Takeuchi, T. & Umezawa, H. 1975 Inhibitory effects of phosphoramidon on neutral metalloendopeptidases and its application on affinity chromatography. Biochemical and Biophysical Research Communications 65 352–357Google Scholar
Laemmli, U. K. & Favre, M. 1973 Maturation of the head of bacteriophage T4. 1. DNA packaging events. Journal of Molecular Biology 80 575–599Google Scholar
Law, B. A. 1979 Reviews of the progress of Dairy Science. Enzymes of psychrotrophic bacteria and their effects on milk and milk products. Journal of Dairy Research 46 573–588Google Scholar
Leinmüller, R. & Christophersen, J. 1982 a [Studies on a heat-resistent proteolytic enzyme system of Pseudomonas fluorescens.] Milchwissenschaft 37 270–272Google Scholar
Leinmüller, R. & Christophersen, J. 1982 b [Studies on the heat inactivation and characterization of termoresistent proteases from Pseudomonas fluorescens.] Milchwissenschaft 37 472–475Google Scholar
Lin, Y., Means, G. E. & Feeney, R. E. 1969 The action of proteolytic enzymes on N, N-dimethyl proteins: basis for a microassay for proteolytic enzymes. Journal of Biological Chemistry 244 789–793Google Scholar
Millière, J. B. & Veillet-Poncet, L. 1979 [Characterization of the proteolytic enzyme systems of 2 psychrotrophic strains isolated from refrigerated raw milk.] Lait 59 269–298Google Scholar
Morihara, K. & Tsuzuki, H. 1978 Phosphoramidon as an inhibitor of elastase from Pseudomonas aeruginosa. Japanese Journal of Experimental Medicine 48 81–84Google Scholar
Richardson, B. C. 1981 The purification and characterization of a heat-stable protease from Pseudomonas fluorescens B52. New Zealand Journal of Dairy Science and Technology 16 195–207Google Scholar
Richardson, B. C. & Te Whaiti, I. E. 1978 Partial characterization of heat-stable extracellular proteases of some psychrotrophic bacteria from raw milk. New Zealand Journal of Dairy Science and Technology 13 172–176Google Scholar
Shaw, B. G. & Latty, J. B. 1982 A numerical taxonomic study of Pseudomonas strains from spoiled meat. Journal of Applied Bacteriology 52 219–228Google Scholar
Stepaniak, L. & Fox, P. F. 1983 Thermal stability of an extracellular proteinase from Pseudomonas fiuorescens AFT 36. Journal of Dairy Research 50 171–184Google Scholar
Stepaniak, L., Fox, P. F. & Daly, C. 1982 Isolation and general characterization of a heat-stable proteinase from Pseudomonas fluorescens AFT 36. Biochimica et Biophysica Acta 717 376–383Google Scholar
Suda, H., Aoyagi, T., Takeuchi, T. & Umezawa, H. 1973 A thermolysin inhibitor produced by Actinomycetes: phosphoramidon. Journal of Antibiotics 26 621–623Google Scholar
Visser, S. 1981 Proteolytic enzymes and their action on milk proteins. A review. Netherlands Milk and Dairy Journal 35 65–88Google Scholar
West, F. B., Adams, D. M. & Speck, M. L. 1978 Inactivation of heat resistant proteases in normal ultrahigh temperature sterilized skim milk by a low temperature treatment. Journal of Dairy Science 61 1078–1084Google Scholar
Zevaco, C. & Desmazeaud, M. J. 1980 Hydrolysis of β-casein and peptides by intracellular neutral protease of Streptococcus diacelylactis. Journal of Dairy Science 63 15–24Google Scholar