02160nas a2200205   4500008004100000022001400041245011400055210006900169260011100238300001200349490000700361520132800368653002001696653002001716653000801736100002801744700002501772700002501797856013201822       2013                        eng d  a0929-866500aPurification and characterization of a thermolysin like protease from thermoactinomyces thalpophilus MCMB-3800 aPurification and characterization of a thermolysin like protease  aEXECUTIVE STE Y-2, PO BOX 7917, SAIF ZONE, 1200 BR SHARJAH, U ARAB EMIRATESbBENTHAM SCIENCE PUBL LTDcAUG  a918-9250 v203 a<p>The extracellular thermolysin like protease (TLP) was purified and characterized from Thermoactinomyces thalpophilus MCMB-380 (Genbank Accession No. EF397000). The enzyme was purified to homogeneity by successive ultra filtration steps using 50 kDa and 10 kDa membrane filters followed by anion exchange chromatography. The molecular mass and isoelectric point of the enzyme were found to be 34.4 kDa and 9.5, respectively. The proteolytic activity was inhibited by EDTA and the enzyme required Ca2+ to show the full activity as well as thermostability. The T-50 of the enzyme at 80 degrees C was 1 h and the activation energy was estimated to be 11.02 Kcal / mol. Atomic absorption spectrophotometric analysis revealed the presence of Zn2+ ion in the protein core indicating that it is a metalloprotease. This protease has commercial potential in catalyzing the condensation reaction of two amino acids for production of the dipeptide aspartame, an artificial sweetener. The one hour time-frame is significantly faster than that of the enzyme thermolysin from Bacillus thermoproteolyticus. Moreover the TLP was stable at 80 degrees C for one hour which makes it industrially robust. The Zn2+ ion in the T. thalpophilus protease appears to be necessary for maintaining the active conformation of the enzyme molecule.</p>10aMetalloprotease10aThermostability10aTLP1 aMajumder, Devipriya, R.1 aKanekar, Pradnya, P.1 aGaikwad, Sushama, M.  uhttps://library.ncl.res.in/content/purification-and-characterization-thermolysin-protease-thermoactinomyces-thalpophilus-mcmb-0