Lopez A, Monsan P
Laboratoire de Génie Biochimique, Institut National des Sciences Appliquées, Toulouse.
Biochimie. 1980;62(5-6):323-9. doi: 10.1016/s0300-9084(80)80161-1.
Dextran sucrase has been produced by fermentation of Leuconostoc mesenteroides NRRL B-512, with and without continuous sucrose addition to improve enzyme production. The enzyme preparation has been concentrated from the fermentation broth by ultrafiltration and purified by gel permeation chromatography on Ultrogel. The specific activity of the dextran sucrase was greatly enhanced by calcium chloride addition to the purified enzyme. This enzyme preparation has been immobilized by covalent coupling onto an amino porous silica support (Spherosil) activated with glutaraldehyde. Immobilized dextran sucrase derivatives with an activity up to 830 dextran sucrase units per g. support could thus be obtained. The effect of the support specific area on coupling efficiency and reaction kinetics has been investigated, and the effect of intraparticular diffusion underlined. The molecular weight distribution of the dextran has been determined when varying several parameters.
通过肠系膜明串珠菌NRRL B - 512发酵生产葡聚糖蔗糖酶,发酵过程中添加或不添加连续蔗糖以提高酶产量。已通过超滤从发酵液中浓缩酶制剂,并通过在Ultrogel上进行凝胶渗透色谱法进行纯化。向纯化的酶中添加氯化钙可大大提高葡聚糖蔗糖酶的比活性。该酶制剂已通过共价偶联固定在经戊二醛活化的氨基多孔二氧化硅载体(Spherosil)上。由此可获得每克载体活性高达830个葡聚糖蔗糖酶单位的固定化葡聚糖蔗糖酶衍生物。研究了载体比表面积对偶联效率和反应动力学的影响,并强调了颗粒内扩散的影响。在改变几个参数时测定了葡聚糖的分子量分布。