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| Recombinant Human Lysozyme LYZL4：Expression in Pichia pastoris and Its Antibacterial Activity |
| LI Xin-xin, TAO Jian-jun, YU Long |
| State Key Laboratory of Genetic Engineering, Institute of Genetics, Fudan University, Shanghai 200433, China |
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Abstract LYZL4 is a kind of c type human lysozyme gene and cloned by our laboratory. It was optimized according to the yeast codon preference. The code sequences optimized LYZL4 gene was cloned under the control of the alcohol oxidase 1 promoter (AOX1) in the vector pPIC9K and integrated into the genome of the methylotrophic yeast Pichia pastoris (P. pastoris) strain GS115 by electroporation. The screening of multiple copies of transformants was operated on YPD medium adding different concentration of G418 geneticin. We could get the multiple copies after screening. After methanol inducing by the flasks, the supernatant was analyzed by SDS-PAGE electrophoresis. The protein bands showed on 14.4 kDa. With human lysozyme (164 071U/mg) as the standard and the red micrococcus as the substrate, The activity of the human lysozyme LYZL4 was detected. The recombinant protein of LYZL4 showed bactericidal activity to micrococcus and its activity was up to 38893U/ml.
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Received: 29 November 2013
Published: 25 January 2014
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