Technical Approach to Simplify the Purification Method and Characterization of Microbial Transglutaminase Produced from Streptoverticillium ladakanum

ABSTRACT: In order to fast and economically purify MTGase from Streptoverticillium ladakanum , a stepwise elution method was developed and compared with linear gradient elution method. MTGase was purified to electrophoretical homogeneity by using CM Sepharose CL‐6B and Blue Sepharose Fast Flow chromatographies by linear gradient or stepwise methods. The recovery of MTGase by linear gradient and stepwise methods were 68.4% and 81.0%, respectively. The optimal temperature and pH were 40 °C and 5.5, respectively. It was stable at pH 5.0 to 7.0 and had a rate constant (KD) of 6.21 °o 10 ‐5 min ‐1 for thermal inactivation at 45 °C. The purified MTGase was activated by K + Na + , Ca 2+ , Mn 2+ , and Mg 2+ , not affected by Fe 3+ , EDTA, but inhibited by Cu 2+ , Zn 2+ , Hg 2+ , Ni 2+ , Co 2+ , Cd 2+ , PCMB, NEM, IAA, and PMSF. A simple stepwise method was developed for the purification of MTGase from S. ladakanum.