【作者】 谭秀山；

【导师】 刘正初； 冯作山；

【作者基本信息】 新疆农业大学 ， 食品科学， 2004， 硕士

【摘要】 本论文对CXJZ95-198菌株在葡萄糖、甘露糖、木聚糖、魔芋粉及果胶五种不同碳源培养基中胞外酶蛋白质总量、还原糖及果胶酶活性的影响进行了研究；对果胶酶、木聚糖酶及β-甘露聚糖酶在甘露糖培养基中的酶活进行了比较；对CXJZ95-198菌株在不同碳源培养基中的发酵液在SDS-PAGE电泳中分泌出的蛋白带作了比较；对CXJZ95-198菌株所产生的胞外酶系中果胶酶进行了分离纯化。在本研究条件下，得到如下结论：(1) CXJZ95-198菌株在以葡萄糖作为碳源的培养基中蛋白质含量变化最大，达到了1.28mg/ml，其次依次为甘露糖、木聚糖、魔芋粉和果胶培养基，分别为1.17 mg/ml，0.94 mg/ml，0.75 mg/ml，0.73 mg/ml；在这五种培养基中，以甘露糖培养基中果胶酶活性最高，达到了79.39 U/ml，其次依次为葡萄糖培养基，魔芋粉培养基，木聚糖培养基，最后为果胶培养基，分别为73.33 U/ml，24.25 U/ml，17.45 U/ml，15.98 U/ml。（2）在甘露糖培养基中，β-甘露聚糖酶活性最高，达到了85.55 U/ml；果胶酶活性次之，为73.39 U/ml；木聚糖酶活性相对较低，有17.91 U/ml。（3）在葡萄糖、甘露糖、木聚糖、魔芋粉及果胶五种不同碳源培养基中的发酵液在SDS-PAGE电泳中分别分泌出32条，33条，35条，36条和21条蛋白带（含亚基）；多数可比较的蛋白质电泳谱带的颜色深浅呈魔芋粉＞甘露糖＞木聚糖＞葡萄糖＞果胶的趋势。（4）确立了以葡萄糖培养基6h的发酵液为提取果胶酶的最初取样点，发酵液经离心，丙酮沉淀，Sephadex G-75葡聚糖凝胶柱层析后获得了凝胶电泳均一的样品，比活力较原酶液提高了9.8倍，回收率达到了32.98%，用SDS-PAGE凝胶电泳测的纯化后的果胶酶分子量为42.2KD。更多还原

【Abstract】 The thesis studied the total exocellular proteins, reductant sugar and the pectinase enzyme activity of CXJZ95-198 strain in glucose, mannose, xylan, mannosan and pectin culture media.Compared enzyme activity of pectinase, xylanose and β-mannanose. The secreted proteins in ferment fluid from different culture media were analysed by SDS-PAGE electrophoresis. A pectinase from CXJZ95-198 was purified. Results have made know: (1) The changes of protein content secreted by CXJZ95-198 in the five media glucose mannose xylan mannosan and pectin were 1.28mg/ml, 1.17mg/ml, 0.94mg/ml, 0.75mg/ml and 0.73mg/ml.The pectinase activity was maximum in the mannose (79.39U/ml) with the order of glucose(73.33 U/ml) mannosan(24.25 U/ml) xylan(17.45 U/ml) and pectin(15.98 U/ml). (2) In mannose medium the β-mannanose activity is the most supreme (85.55 U/ml), the prectinase activity(73.39 U/ml) is take second place，the xylanose activity(17.91 U/ml) is third. (3) By the analysis of SDS-PAGE electrophoresis, CXJZ95-198 secrets exocellular proteins bands (include protein subunit) in media of glucose, mannose ,xylan, mannosan and pectin respective were 32 bands, 33 bands,35 bands, 36 bands and 21 bands respectively . The colour of most detected bands became light in the five media with the order of mannosan, mannose, xylan, glucose and pectin. (4)The pectinase in the supernatant was purified by centrifuge , acetone precipitation and sephadex G-75 cloumngel filtration. The specific activity was increased 9.8-fold with an activity recovery of 32.98% .The molecular weight of pectinase was 42.2KD estimated by SDS-PAGE.更多还原