Study On The Fermentation Conditions And Enzymatic Characteristics Of Laccase From Trametes Sanguinea

In this study,the conditions of laccase production of liquid and solid fermentation were optimized and the enzymatic properties of the enzyme were studied by using trametes sanguinea as material.The aim of this study was to optimize the fermentation process of trametes sanguinea laccase,improve the production of laccase,and lay a foundation for further development and application of trametes sanguinea laccase.The following results were obtained through the experiment:（1）The conditions of the liquid fermentation of the trametes sanguinea were optimized,the ingredients of the medium were optimized by single factor test,and then the concentration of medium component and the environment of liquid culture were optimized by orthogonal test.the optimized medium was glucose 15g,sucrose15g,peptone 2g,ammonium2g,potassium dihydrogen phosphate two3g,magnesium1.5g,copper2mmol/L.Thesuitable liquid fermentation environment condition is:the pH is 5,the liquid volume is 110mL,the inoculation quantity is 4%,temperature set to28℃,the ferment liquid laccase activity is 163u/m L,the mycelium extracts the crude enzymatic enzyme activity to be the 150.37u/mL,3 times times higher than before optimization.（2）The effects of three kinds of lignocellulose,such as sawdust,cottonseed Peel and corncob on the laccase production by liquid fermentation of the trametes sanguinea were studied.The results showed that the activity of sawdust and cottonseed was the highest in the seventh day,and the activity of laccase in the corncob was highest in the sixth day.Through comparison and analysis,it is found that the complete culture medium enzyme activity of sawdust is higher than the other two,so the ability of sawdust to induce laccase secretion is stronger than that of cottonseed skin and corncob.（3）The medium of solid fermentation of trametes sanguinea and the addition of different lignin cellulose tablets were optimized,the results showed that the plate medium containing sawdust has the highest activity of producing enzymes,optimized selection of the optimal solid fermentation medium for sawdust 58%,cottonseed Peel10%,corncob 10%,wheat bran 20%,gypsum 1%,lime 1%,cupric sulfate 1.5mmol/l,the enzyme activity was 202.22U/mL。（4）The research results of enzymatic properties showed that,the optimum reaction temperature of the laccase was 37℃,and the laccase under 50℃had good stability,Cu²⁺、Zn²⁺、Mg²⁺、K⁺、Na⁺can promote the activity of the enzyme,in which Cu²⁺promotes the action,while Fe³⁺、Mn²⁺inhibit the enzyme activity,with ABTS as the substrate,the Km value is 0.095mmol/l,The low Km value of the laccase indicated that the enzyme had higher affinity to substrate abts.