Studies On 19 Species Of Magnolia Plant Peroxidase

Magnoliaceae belongs to the original group of angiosperms.Abundant species of Magnoliaceae from china provide precious materials for the study of the origin and phylogeny of angiosperms.The POD isozymes as a molecular marker can reflect the variation of species during evolution,which is encoded by conserved polygenes.In this study,the differences in the POD isozyme spectrum of 19 species of Magnoliaceae plants were studied by using the aluminum lactate acid electrophoresis,peroxidase isozyme patterns and cluster analysis techniques,which of those provide more technical support for species diversity and relationship identification of Magnolia plants.At the same time,it was found that most Magnolia plants have high POD activity in their leaves during the study of POD isozyme diversity.The purpose of this paper is separate and purify the POD from the leaves of Michelia maudiae Dunn,which is a common plant in the Enshi area.We also have studied its partial enzymatic properties,and discussed the characteristics of its removal of organic pollutants BPA.We immobilized POD from the leaves of Michelia maudiae Dunn by absorbing and crosslinking method with Fe₃O₄-Sodium Alginate to increase the utilization efficiency of the enzyme.This research can provide theoretical and technical support for the rational use of Magnolia plant resources.The results were summarized as follows:?1?The aluminum lactate acid electrophoresis can successfully separate the POD isozyme from the leaves of Magnolia plants,but other electrophoresis approaches,like conventional alkaline and potassium acetate acid electrophoresis,cannot separate it effectively.The results of cluster analysis reveal that 19 species of Magnolia plants were clustered into four categories,which were basically consistent with the classification of19 plants in the Flora of China.Indicating that this method of electrophoretic separation and cluster analysis can be applied to POD isozyme diversity research of Magnolia plants.?2?After buffer extraction,ATPE decolorization,and DEAE-Sepharose FF ion-exchange chromatography,the purification multiple of BPOD was 5.5-fold,the relative of SPOD was 55.2-fold,and the yield of enzyme activity was 19%.The results of enzymatic properties represent that the optimum reaction temperature of this enzyme was 50°C,and it could be stored stably in buffer when the temperature is below 50°C,reflecting good thermal stability.Its optimum pH was 4.5.When the pH was in the range of 5.5 to 9.5 for 12 hours,the enzyme activity did not decrease significantly,indicating that the enzyme has good pH stability.?3?The results of the study on the removal of BPA by POD from the leaves of Michelia maudiae Dunn showed that the removal rate of BPA was more than 90%under the conditions of pH 4⁷,30⁴0°C and reach the molar ratio of H₂O₂/BPA to 0.8 in the removal.The POD from the leaves of Michelia maudiae Dunn has a wider pH and temperature adaptation range and a higher clearance rate of BPA by comparison with PODs such as HRP,TP,and SBP.M.maudiae Dunn leaves peroxidase had a convincing future in researches of phenol degradation.?4?The results of the immobilization of POD from the leaves of Michelia maudiae Dunn demonstrate that the absorbing and crosslinking method with Fe₃O₄-Sodium Alginate has a higher enzyme immobilization rate than the sodium alginate embedding method and can increase the enzyme immobilization rate by 14.2%.Some of the properties of the immobilized enzyme obtained by adsorption cross-linking method of Fe₃O₄-Sodium Alginate showed that the immobilized enzyme prepared by this method could maintain relatively high enzyme activity at room temperature or even at low temperature due to the addition of Fe₃O₄.The optimum pH for the reaction was lower than the free enzyme by 1.0,making it able to adapt to a wider range of acidic pH.When the immobilized enzyme was recycled for 5 times,the enzyme activity was reduced 55%,indicating that the immobilized enzyme can be used repeatedly and has good operational stability.