黄绿木霉固定化生产纤维素酶及酶学特性的研究  

Studies on Cellulase Production by Immobilized Cells and Characteristics of Cellulase

作　　者：孙冬梅[1] 杨谦[1] 宋金柱[1] 陈忠祥[1] 

机构地区：[1]哈尔滨工业大学生命科学与工程系

出　　处：《林产化学与工业》2006年第2期79-82,共4页Chemistry and Industry of Forest Products

摘　　要：利用海藻酸钙凝胶包埋法制备固定化细胞黄绿木霉,结果表明：海藻酸钠质量浓度 50 g/L,培养基初始pH值4.0～4.5,以滤纸浆为培养基碳源培养的小球的稳定性好.发酵产酶培养时以滤纸浆为碳源,海藻酸钠质量浓度为 50 g/L, CaCl2质量浓度控制在 20 g/L,培养基初始pH值选择为4.5,产酶效果好、酶活力高、保持时间长.添加表面活性剂Twin-80后,产酶能力可进一步提高.通过（NH4）2SO4分级沉淀、Sephadex G-100分子筛层析和DEAE Sephadex A-50离子交换层析等步骤,分离纯化出黄绿木霉纤维素酶系中达到电泳纯的3种内切葡聚糖酶（EGⅠ、EGⅡ、EGⅢ）和2种β-葡萄糖苷酶（BGⅠ、BGⅡ）.通过SDS-PAGE和IEF电泳测得5个酶组分的相对分子质量（Mr）分别为62 300、 71 900、 52 600、 85 300和 78 300,等电点分别为5.4、 4.8、 5.0、 5.6和5.8.Immobilized cells were prepared through embedment of Trichoderma aureoviride with calcium alginate. The suitable culture conditions were sodium alginate 50 g/L, initial pH value of the substrate 4.0-4.5 and using filter paper pulp as carbon source of the substrate. The enzymatic activity of the immobilized cells was improved when concentration of calcium dichloride was 20 g/L, initial pH value of the substrate was 4.5, concentration of sodium alginate was 50 g/L using filter paper pulp as carbon source of the substrate. Addition of the surfactant Twin-80 could further increase enzymatic activity. Three endoglucanases and two β-glucosidases from the fungi were separated and purified to electrophoretic homogeneity through processes of （NH4 ）2SO4 fractional precipitation, Sephadex G-100 gel chromatography and DEAE Sephadex A-50 ion exchange chromatography. They were designated as EGⅠ , EGⅡ , EGⅢ, BG Ⅰ and BGⅡ , with relative molecular weights （Mr）of62 300, 71 900, 52 600, 85 300 and 78 300 by SDS-PAGE, and isoelectric points of 5.4, 4.8, 5.0, 5.6 and 5.8, respectively.

关 键 词：纤维素酶 黄绿木霉 固定化 等电点 

分 类 号：TQ351.011[化学工程] Q814.2[生物学—生物工程]