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| Optimization of Protease Inhibitor Mixture for Protein Extraction from Cells in Proteomics Study of Streptomyces avermitilis |
| GE Wei-feng1, CHU Xiao-he2, WANG Yong-hong1, ZHANG Si-liang1 |
1. State Key Laboratory of Bioreactor Engineering, East China University of Science & Technology, Shanghai 200237,China;
2. Zhejiang Shenghua Biok Biology CO, LTD., Huzhou 313220,China |
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Abstract A lot of proteases are synthesized for cell growth and metabolism by Streptomyces avermitilis during growing in the complex medium. However, the 2-DE protein sample preparation for intracellular proteomics analysis of S. avermitilis became difficult because of the hydrolysis of protein by massive proteases when cells were disrupted. An effective protease inhibitor mixture was obtained by single factor and orthogonal optimum design based on the knowledge of intracellular protease composition of S. avermitilis. By verifying expriments, it was validated that the protease inhibitor mixture can effectively protect large molecular weight proteins from enzymolysis during sample preparation.
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Received: 24 August 2010
Published: 25 December 2010
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