Identification in Tetrahymena pyriformis of 3-hydroxy-3-methyl glutaryl coenzyme a lyase: its purification and properties |
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| Authors: | P Prasanna C E Holmlund |
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| Institution: | 1. Posgrado en Recursos Genéticos y Productividad-Ganadería, Colegio de Postgraduados Campus Montecillo, Km 36.5 Carretera México-Texcoco, Montecillo, Texcoco, Estado de México 56230, Mexico;2. Posgrado de Innovación en Manejo de Recursos Naturales, Colegio de Postgraduados Campus San Luis Potosí, Iturbide 73, Salinas de Hidalgo, San Luis Potosí 78620, Mexico;3. Posgrado en Estrategias para el Desarrollo Agrícola Regional, Colegio de Postgraduados Campus Puebla, Boulevard Forjadores de Puebla No. 205, Santiago Momoxpan, Municipio San Pedro Cholula, Puebla 72760, Mexico;1. State Key Laboratory of Environmental Criteria and Risk Assessment, Chinese Research Academy of Environmental Sciences, Beijing, 100012, China;2. College of Environment, Hohai University, Nanjing, 210098, China;3. Key Laboratory of Drinking Water Science and Technology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing, 100085, China;4. China National Environmental Monitoring Centre, Beijing, 100012, China;1. Faculty of Agronomy and Forestry Engineering, Eduardo Mondlane University, Mozambique;2. Faculty of Agronomic and Biological Sciences, Púnguè University, Mozambique |
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| Abstract: | The major HMG-CoA utilizing enzyme activity in T. pyriformis has been determined to be HMG-CoA lyase. The enzyme was purified 32-fold to a specific activity of 431 units/mg from a mitochondrial fraction. Sephacryl S-200 chromatography gave an estimated molecular weight of 50,000 daltons for the HMG-CoA lyase. SDS gel electrophoresis revealed two bands stained by Coomassie Blue--a major band of 50,000 daltons and a minor band of 25,000 daltons. The latter is believed to be an impurity in the preparation. The enzyme has a pH optimum of 9.0, is stimulated slightly by sulfhydryl reagents, and requires a divalent cation for maximum activity. The KM for HMG-CoA is 15 microM. |
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