Heat Stable Enzymes from Thermophiles
Abstract
Alkaline phosphatase is widely used in the military and civilian sectors. Commercially available enzyme from calf intestine is the weak link in many applications, breaking down when exposed to elevated temperatures or after being stored for prolonged periods. An extremely heat stable alkaline phosphatase produced by a thermophilic bacterium had been previously identified. The major deficiency was a low specific activity. Research described in this report focused on increasing the specific activity of the heat stable enzyme. This was successfully accomplished by cloning the alkaline phosphatase gene from the wild type into E. coli. One 40-kd cloned product was purified to near homogeneity by nickel affinity chromatography of the N-terminal histidine tagged protein. Purified, cloned alkaline phosphatase exhibited good activity and was thermally stable.
Document Details
- Document Type
- Technical Report
- Publication Date
- Feb 01, 1998
- Accession Number
- ADA342616
Entities
People
- Fred Albert
- Joan Combia
- Karen Budwill
- Kenneth Runnion
- Michael Williamson
Organizations
- Edgewood Chemical Biological Center