Isolation of latent phenoloxidase from prepupae of the housefly,Musca domestica |
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Affiliation: | 1. State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing, China;2. University of Chinese Academy of Sciences, Beijing, China;1. Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Sciences, Central China Normal University, Wuhan 430079, China;2. School of Biological Sciences, University of Missouri, Kansas City, MO 64110, USA;1. Department of Green Material Technology, Green Technology Research Institute, Chinese Petroleum Corporation(CPC Corporation), Kaohsiung, Taiwan;2. Department of Mechanical and Automation Engineering, I-Shou University, Kaohsiung, Taiwan;3. Department of Fragrance and Cosmetic Science, Kaohsiung Medical University, Kaohsiung, Taiwan |
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Abstract: | Latent phenoloxidase was purified from prepupae of the housefly, Musca domestica vicina Maquart. The purification procedures included DEAE-cellulose column chromatography, sucrose density gradient centrifugation adn second sucrose density gradient centrifugation. The final preparations appear to be homogeneous based on results obtained from polyacrylamide gel electrophoresis in the presence of EDTA. Electrophoresis in the absence of EDTA caused spontaneous activation of latent phenoloxidase. While latent phenoloxidase was fairly stable over the range of temperatures between 0 and 40°C, it was quite sensitive to changes in pH, being stable only around pH 6.0. The molecular weight of latent phenoloxidase was estimated to be 178,000, as determined by gel filtration and sucrose density gradient centrifugation. Furthermore, phenoloxidase formed by the activation of latent phenoloxidase indicated a higher molecular weight (340,000) than that of latent phenoloxidase. Thus, it appears that the mechanism of the activation of latent phenoloxidase involves the association and disassociation system. |
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