传统的纯化微生物产几丁质酶的方法步骤繁琐、不易控制、得率较低。经过实验探索得出一种较简单、快速、并可批量处理的几丁质酶纯化法。将含有几丁质酶的发酵上清液与胶体几丁质混合后离心,几丁质酶随胶体几丁质沉降而与其他成分分离,用蒸馏水洗涤沉淀,再用蒸馏水重悬,放入30℃恒温箱温育72 h,胶体几丁质被完全水解,从而得到纯几丁质酶。此纯化方法条件温和,可保证纯化后的酶保持其生物活性。SDS-PAGE检测及比色法对其纯度检测表明几丁质酶纯化回收率达到88.53%。

Traditional purification of chitinase from the fermentation supernatant had some weaknesses such as complicated steps,difficult in controlling and low recovery ratio.Then we took experimental exploration and find a simple,fast and convenient method in the process of chitinase purification.This method may also be used in Batch processing.Detailed method is: fermentation supernatant which containing chitinase was first mixed with colloidal chitin,after centrifugation,the chitinase and the colloidal chitin precipitated and separated with other components in the supernatant.Then the precipitated was washed by distilled water for several times,the precipitate was then resuspended in distilled water and then sterilized at 30 ℃for 72h.After the colloidal chitin was completely hydrolyzed,chitin enzyme was purified and Abstracted.This method requires simple,mild purification conditions,ensuring the purified enzyme maintain its biological activity.SDS-PAGE and colorimetry detecting methods indicated that the recovery rate of chitinase using this method can reach 88.53%.