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. 1983 Aug;46(2):333–337. doi: 10.1128/aem.46.2.333-337.1983

Heat-stable protease from Pseudomonas fluorescens T16: purification by affinity column chromatography and characterization.

T R Patel, D M Jackman, F M Bartlett
PMCID: PMC239382  PMID: 6414369

Abstract

A heat-stable extracellular protease from Pseudomonas fluorescens T16, a psychrotroph, was purified by affinity column chromatography on a carbobenzoxy-D-phenylalanine-triethylene tetramine-Sepharose-4B column. The purified enzyme is a monomer with a molecular weight of 38,905 +/- 2,000. In an analytical ultracentrifuge, the Schlieren profile revealed a single symmetrical peak. The sedimentation coefficient was estimated to be 3.93S. Alpha-casein was the preferred substrate, with a Km of 0.05 mM. Heating crude enzyme and purified enzyme in buffer at 50, 90, and 120 degrees C resulted in a rapid initial loss of more than 50% of the initial activity followed by a gradual inactivation which exhibited first-order kinetics. The activation energy for the hydrolysis of casein was calculated to be 3.2 kcal/mol (13.4 kJ/mol).

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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