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运用从自然界中分离得到的一株具有产生特效高活性碱性果胶酶的软腐欧文氏菌株ErwiniaCarotovoraIFO 3830 ,通过发酵得到了含有高活性碱性果胶酶的发酵液 .由于果胶酶在该发酵液中浓度极低 ,仅为数 μg/g左右 .为了从大量的但含有目的酶浓度却极低的溶液中将目的酶分离出来 ,一般所使用的化工浓缩方法由于其操作温度较高使酶产品失活而无法运用 .目前被普遍用于酶浓缩的硫酸铵沉淀法 ,不仅化学药品的消耗极大 ,而且后处理工艺又比较复杂 ;回收得到的沉淀酶制品中因混入了大量的培养基和其他杂质 ,导致后处理工序的更加复杂 .本研究采用截留分子量为 5 0 0 0和 10 0 0 0 (道尔顿 )的超滤平面膜组件 ,可以直接从去除了菌体的发酵液中浓缩回收了目的产物碱性果胶酶 ,在浓缩率在 2 0倍的条件下 ,取得了98.3%的高回收率 .此外 ,还对超滤过程的表观阻力、浓缩率、渗透流束和浓缩液浓度等随着超滤过程的进行的变化规律进行了较为详细的研究
Fermentation broth containing highly active alkaline pectinase was obtained by fermentation using Erwinia carotovora IFO 3830, an Erwinia carotovora IFO 3830 strain isolated from nature, producing pectinase with high activity. The concentration of the fermentation broth is very low, only a few μg / g or so in order to isolate the enzyme of interest from a large number of solutions containing a very low concentration of the target enzyme, the chemical concentration method generally used due to its high operating temperature so that the enzyme The product is inactivated and can not be used.At present, ammonium sulfate precipitation method commonly used for enzyme concentration not only consumes a great deal of chemicals, but also the post-treatment process is rather complicated; the precipitated enzyme product recovered contains a large amount of medium And other impurities, leading to more complex post-treatment process.In this study, the molecular weight cut-off of 50000 and 1000 (Dalton) ultrafiltration planar membrane module can be directly removed from the cell culture broth The target product alkaline pectinase was recovered by concentration, and a high recovery rate of 98.3% was obtained at a concentration of 20 times. In addition, the apparent resistance of the ultrafiltration process, concentration , Etc. permeate stream concentration and concentration variation with the ultrafiltration process will be carried out more detailed studies