Technical Approach to Simplify the Purification Method and Characterization of Microbial Transglutaminase Produced from Streptoverticillium ladakanum |
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Authors: | M.-L. Ho S.-Z. Leu J.-F. Hsieh S.-T. Jiang |
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Affiliation: | Authors are with the Dept. of Food Science, National Taiwan Ocean University, Keelung, Taiwan 202, R.O.C. |
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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+, 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 stepwise method was developed for the purification of MTGase from S. ladakanum. |
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Keywords: | MTGase purification method properties of MTGase |
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