变色栓菌产漆酶培养条件优化及酶学特性研究

Research on the Conditions Optimization for Laccase Production from Trametes Versicolor and the Enzymatic Characteristics on Laccas

对筛选到的变色栓菌（mys 004）产漆酶液体培养方式、培养基组成和培养条件进行了优化,并对其酶学特性进行了研究;确定了培养基组成及培养条件：20%马铃薯汁1 L、K2HPO41.0 g、NaCl 0.5 g、MgSO4.7H2O1.5 g、NaNO32.5 g、CaCl2.2H2O 0.1 g、FeCl30.02 g、吐温80 1.2 mL、麦麸10 g、愈创木酚0.006%,pH 4.5,28℃,200 r/min,培养7 d,酶活达到751 U/mL。该漆酶的适宜反应温度范围为30~55℃,最适50℃;最适反应pH 4.5;贮存温度不能超过60℃,以低于40℃为最佳;含量为5 mmol/L的Cu2＋、Mg2＋对酶活没影响;Zn2＋、Mn2＋、Co2＋、Al3＋都可使酶活丧失50%左右,Fe3＋可使酶活丧失70%以上,Fe2＋可使酶活丧失为零。

The constituent of culture medium and culture conditions for the trametes versicolor （mys 004） screened was studied and optimized. And its enzymatic characteristics were also researched. Its culture medium consisted of 20% potato juice 1L（ K2HPO4 1.0 g, NaC10.5 g, MgSO4·TH201.5 g, NaNO32.5 g） CaCl2·2H2O 0.1 g, FeCl3 0.02 g, tween 80 1.2 mL, pollard 10 g, guaiacol 0.006%. The activity of the yielding laccase was 751 U/mL under the conditions of the culture pH 4.5, the culture temperature 28℃, the rotational speed of culture 200 r/min, the culture time 7 d. Its enzymatic characteristics was using the temperature 30-55℃ （the optimum temperature 50 ℃）, the optimum pH 4.5. The storage temperature could not exceed for 60℃, the optimum temperature less than 40℃. When there was 0.5 mmol/L Cu^2＋ or Mg^2＋, the enzymatic activity was not effected. When there was 5 mmol/L Zn^2＋, Mn^2＋, Co^2＋ or A1^3＋, the enzymatic activity was descended approximately for 50%. The enzymatic activity was descended approximately for 70% if the Fe^3＋ existed. The enzymatic activity entirely lost if the Fe^2＋ existed.