This experiment passing to grope for the carbon source constitutes of the culture medium and using t . reesei rut c - 30 induced the expression of # - mannanase ( # - 1 , 4 - mannan mannohydrolase ec 3 . 2 . 1 . 78 ) . in this experiment i put the constant carbon source ( lactose and locust bean gum ) in the foundation culture medium ( mandels nourishment liquid ) of t . reesei rut c - 30 , then proceeded the variable carbon source ( dragon spruce fiber , com rush pith fiber , wheat straw fiber , wheat straw xylan , corn rush pith xylan , dragon spruce mannan ) to single factor , double factor , three factor , four factor and five factor orthogonal experiment . 1 determined the activity of p - mannanase using locost bean gum as substract by the 3 , 5 - dinitosalicylic acid method , and observed the growing situation of the gernic at the end i selected the directions for the inducement expression of the # ? mannanase from trichoderma reesei rut - c30 that contained the dragon spruce fiber , wheat straw xylan , dragon spruce mannan 在里氏木霉rutc - 30的基础培养基( mandels营养液)中加入固定碳源乳糖和槐豆胶,然后将可变碳源(云杉纤维、玉米芯纤维、麦杆纤维、麦杆木聚糖、玉米芯木聚糖、云杉甘露聚糖)进行单因子、双因子、三因子、四因子、五因子的里氏木霉rutc - 30正交培养实验,并以槐豆胶为底物用3 , 5二硝基水杨酸法测定培养液中?甘露聚糖酶的活力。从而确定了酶活最高且菌体生长良好的含云杉纤维、麦杆木聚糖和云杉甘露聚糖的诱导培养基为最佳培养基,用该培养基培养的里氏木霉( t . reesei ) rutc - 30使其转录的-甘露聚糖酶( - 1 , 4 - mannanmannohydrolaseec3 . 2 . 1 . 78 ) mrna量能够满足rt - pcr的要求。
2.
The page revealed the culture supernatant of the initial strain and the mutant contained different protein bands , which exactly demonstrated at protein level that a . niger j 506 was surely a mutant of a . niger m1 . zymogram stained with xylan - remazol brilliant blue for detecting xylanase showed there are three different xylanases in the mutant culture , while two xylanases in initial strain . what is important , the third xylanase in a . niger j 506 have higher activity and more production levels from page and zymogram of xylanase 尤其是在木聚糖酶谱带检测中发现,突变株发酵液中有三种类型的木聚糖酶,而出发菌株中只有两种类型的木聚糖酶,并且通过考马斯亮蓝g250染色和琼脂糖板上的透明圈发现,突变株中第三种类型的木聚糖酶不仅表达量很大,活力也很高。