Abstract
Pseudomonas fluorescens RO98, a raw milk isolate, was inoculated into McKellar's minimal salts medium and incubated at 25°C for 48 h to allow production of protease. A zinc-metalloacid protease was purified from the cell-free concentrate by anion exchange and gel filtration chromatography. The purified protease was active between 15 and 55°C, and pH4.5 and 9.0, and was stable to pasteurization. The enzyme had pH and temperature optima for activity of 5·0 and 35°C, respectively. It was heat stable with a D55 of 41 min and a D(62·5) of 18 h. Molecular weight of the enzyme was estimated to be 52 kDa by SDS PAGE and size exclusion chromatography. Values for k(M) of 144·28, 18·73, 110·20 and 35·23 μmol were obtained for whole, α-, β- and κ-casein, with a V(max) of 8·26, 0·09, 0·42 and 0·70 μmol mg-1 min-1, respectively. The enzyme hydrolysed κ-casein preferentially when incubated with artificial casein micelles.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 280-288 |
| Number of pages | 9 |
| Journal | Journal of Applied Microbiology |
| Volume | 89 |
| Issue number | 2 |
| DOIs | |
| State | Published - 2000 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Agricultural and Biological Sciences(all)
- Applied Microbiology and Biotechnology
- Biotechnology
- Microbiology
Cite this
Isolation and characterization of a protease from Pseudomonas fluorescens RO98. / Koka, R.; Weimer, Bart C.
In: Journal of Applied Microbiology, Vol. 89, No. 2, 2000, p. 280-288.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Isolation and characterization of a protease from Pseudomonas fluorescens RO98
AU - Koka, R.
AU - Weimer, Bart C
PY - 2000
Y1 - 2000
N2 - Pseudomonas fluorescens RO98, a raw milk isolate, was inoculated into McKellar's minimal salts medium and incubated at 25°C for 48 h to allow production of protease. A zinc-metalloacid protease was purified from the cell-free concentrate by anion exchange and gel filtration chromatography. The purified protease was active between 15 and 55°C, and pH4.5 and 9.0, and was stable to pasteurization. The enzyme had pH and temperature optima for activity of 5·0 and 35°C, respectively. It was heat stable with a D55 of 41 min and a D(62·5) of 18 h. Molecular weight of the enzyme was estimated to be 52 kDa by SDS PAGE and size exclusion chromatography. Values for k(M) of 144·28, 18·73, 110·20 and 35·23 μmol were obtained for whole, α-, β- and κ-casein, with a V(max) of 8·26, 0·09, 0·42 and 0·70 μmol mg-1 min-1, respectively. The enzyme hydrolysed κ-casein preferentially when incubated with artificial casein micelles.
AB - Pseudomonas fluorescens RO98, a raw milk isolate, was inoculated into McKellar's minimal salts medium and incubated at 25°C for 48 h to allow production of protease. A zinc-metalloacid protease was purified from the cell-free concentrate by anion exchange and gel filtration chromatography. The purified protease was active between 15 and 55°C, and pH4.5 and 9.0, and was stable to pasteurization. The enzyme had pH and temperature optima for activity of 5·0 and 35°C, respectively. It was heat stable with a D55 of 41 min and a D(62·5) of 18 h. Molecular weight of the enzyme was estimated to be 52 kDa by SDS PAGE and size exclusion chromatography. Values for k(M) of 144·28, 18·73, 110·20 and 35·23 μmol were obtained for whole, α-, β- and κ-casein, with a V(max) of 8·26, 0·09, 0·42 and 0·70 μmol mg-1 min-1, respectively. The enzyme hydrolysed κ-casein preferentially when incubated with artificial casein micelles.
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UR - http://www.scopus.com/inward/citedby.url?scp=0033864676&partnerID=8YFLogxK
U2 - 10.1046/j.1365-2672.2000.01108.x
DO - 10.1046/j.1365-2672.2000.01108.x
M3 - Article
C2 - 10971760
AN - SCOPUS:0033864676
VL - 89
SP - 280
EP - 288
JO - Journal of Applied Microbiology
JF - Journal of Applied Microbiology
SN - 1364-5072
IS - 2
ER -