Purification and properties of cold-active metalloprotease from Curtobacterium luteum and effect of culture conditions on production.
- Author:
Mohammed KUDDUS
1
;
Pramod W RAMTEKE
Author Information
1. Protein Research Laboratory, Department of Biotechnology, Integral University, Kursi Road Lucknow 226026, India. kuddus_biotech@yahoo.com
- Publication Type:Journal Article
- MeSH:
Actinomycetales;
enzymology;
growth & development;
Cold Temperature;
Culture Media;
Enzyme Stability;
Hydrogen-Ion Concentration;
Metalloproteases;
isolation & purification;
metabolism;
Soil Microbiology
- From:
Chinese Journal of Biotechnology
2008;24(12):2074-2080
- CountryChina
- Language:English
-
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.
