Technical approach to simplify the purification method and characterization of microbial transglutaminase produced from Streptoverticillium ladakanum

In order to fast and economically purify MTGase from Streptoverticillium la dakanum, a stepwise elution method was developed and compared with linear g radient elution method. MTGase was purified to electrophoretical homogeneit y by using CM Sepharose CL-6B and Blue Sepharose Fast Flow chromatographies by linear gradient or stepwise methods. The recovery of MTGase by linear g radient and stepwise methods were 68.4% and 81.0%, respectively, The optima l temperature and pH were 40 degrees C and 5.5, respectively. It was stable at pH 5.0 to 7.0 and had a rate constant (K-D) of 6.21 x 10(-5) min(-1) fo r thermal inactivation at 45 degrees C. The purified MTGase was activated b y K+, Na+, Ca2+, Mn2+, and Mg2+, not affected by Fe3+, EDTA, but inhibited by Cu2+, Zn2+, Hg2+, Ni2+, Co2+, Cd2+, PCMB, NEM, IAA, and PMSF. A simple s tepwise method was developed for the purification of MTGase from S. ladakan um.