Purification and properties of cold-active metalloprotease from Curtobacterium luteum and effect of culture conditions on production

Sheng Wu Gong Cheng Xue Bao. 2008 Dec;24(12):2074-80.

Abstract

Curtobacterium luteum, a gram-positive psychrotrophic bacterium, secreting an extracellular protease was isolated from the soil of Gangotri glacier, Western Himalaya. The maximum enzyme production was achieved when isolate was grown in a pH-neutral medium containing skim milk at 15 degrees C over 120 hour. The metal ions such as Zn2+ and Cr2+ enhanced enzyme production. The specific activity of purified enzyme was 8090 u/mg after 34.1 fold purification. The 115 kD enzyme was a metalloprotease (activity inhibited by EDTA and EGTA) and showed maximum activity at 20 degrres C and pH 7. The enzyme was active over a broad pH range and retained 84% of its original activity between pH 6-8. There was no loss in enzyme activity when exposed for 3 hours at 4 degrees C-20 degrees C. However, lost 65% of activity at 30 degrees C, and was almost inactivated at 50 dgrees C, but was resistant to repeated freezing and thawing. The enzyme activity was stimulated by manganese ions; however, it was inactivated by copper ions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actinomycetales / enzymology*
  • Actinomycetales / growth & development
  • Cold Temperature*
  • Culture Media
  • Enzyme Stability
  • Hydrogen-Ion Concentration
  • Metalloproteases / isolation & purification*
  • Metalloproteases / metabolism
  • Soil Microbiology*

Substances

  • Culture Media
  • Metalloproteases