黄海黄杆菌YS-9412-130低温碱性蛋白酶的制备工艺研究

The study of preparation technology of low-temperature alkaline protease from Flavobacterium YS-9412-130

对黄海黄杆菌所产低温碱性蛋白酶的工业生产技术和电泳纯级试剂酶的分离纯化工艺进行了实验。按照设计的中试生产技术 ,每吨发酵液可得酶粉 2 0 kg,粗酶比活大于 2 0× 10 4 U/ g;同时运用一系列纯化手段和条件 ,制备出电泳纯级试剂酶 ,纯化过程蛋白回收率在 7%左右 ,活性回收率在60 %以上 ,比活性也稳定在 1.9× 10 3U/ mg以上 ,激光灰度扫描结果试剂酶纯度大于 99% ,可以作为试剂酶满足进一步生物学研究和实际应用。其产品质量和制造工艺均达到了中试放大规模的要求。

A preliminary study on industrial abstraction technology and purification technique of the low temperature alkaline protease from Flavobacterium YS 9412 130 is carried out with emphasis on some universal questions. Based on the predesigned pilot technology, 20 kg enzyme powders can be obtained from one ton fermented broth and the specific activity of crude enzyme is over 20×10 4U/g. Meanwhile,the enzyme electrophoreticly pure is prepared through a series of procedures of purification mentioned above. Through the purification course, the recovery rate of protein is about 7%, the recovery rate of activity over 60%, the specific activity over 1 9×10 3U/mg. The purity of reagent enzyme is more than 99% in laser grey scaning. It can be used as reagent enzyme for further biologic research and actual application. The product quality and the manufacturing technique meet the requirements of pilot production.