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| Large Scale Purification of Pharmaceutical Grade Microbial Transglutaminase from Streptoverticillium mobaraense |
| PEI Zheng-pei1, LI Bo2, ZHANG Wei1, GAO Hong-liang1, CHANG Zhong-yi1, JIN Ming-fei1, LU Wei3, BU Guo-jian3 |
1. School of Life Science of East China Normal University, Shanghai 200241, China;
2. Shanghai Vocational Management College of Trade Union, Shanghai 201415, China;
3. Taixing Dongsheng Food and Technology Limited Company, Taixing 225411, China |
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Abstract A safe efficient microbial transglutaminase purification method applied for industrial manufacture was introduced in order to improve the purity of transglutaminase and extend its applications for pharmaceuticals. Cultures was centrifuged at 10 000 r/min 4℃ to remove the cells before adjusting conductivity to 4.1 mS/cm and pH to 5.5 for purification. The sample was purified by cation-exchange chromatography at a linear velocity of 60 cm/h in the first step. In this step SP Sepharose FF as a packing provided high selectivity and high loading for the target protein. After primary purification, the sample was further processed/isolated with hydrophobic chromatography packed with phenyl Sepharose 6 FF (high sub). SDS-PAGE pattern manifested a purity of over 95% and HPLC analysis indicated a purity of above 99% were achieved after this two purification step method. LPS of the purified sample was determined at 0.013 EU/ml with tachypleus amebocyte lysate, met the blood products requirement (<0.15 EU/ml) provided in Chinese Pharmacopoeia. Thus the method described/established is efficient and practical in microbial transglutaminase purification.
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Received: 14 February 2014
Published: 25 April 2014
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