壳聚糖酶的分离纯化及性质研究  

Study on Purification and Properties of Chitosanase

作　　者：袁建平[1] 侯晓强[1] 

机构地区：[1]廊坊师范学院生命科学学院,河北廊坊065000

出　　处：《湖北农业科学》2013年第15期3647-3649,共3页Hubei Agricultural Sciences

基　　金：国家自然科学基金青年科学基金项目(31100314);河北省科技支撑计划项目(11220931)

摘　　要：利用芽孢杆菌(Bacillus sp.)Yg菌株发酵产壳聚糖酶,采用分段醇析法分离纯化发酵产物,通过不连续SDS-PAGE法测定壳聚糖酶相对分子质量。结果表明,发酵产物先用体积分数40%的乙醇初步醇析,离心去除沉淀,而后加乙醇至体积分数为60%再次醇析,所得沉淀冷冻干燥即为壳聚糖酶,所得酶的回收率和纯度均较高。所得壳聚糖酶的相对分子质量约为41.7 ku,最适反应温度为45℃,最佳反应pH为5.5。该酶在4℃条件下保存稳定性较好,在65℃及以上温度下短时间内即全部失活。Chitosanase was produced by fermentation of Bacillus sp. Yg strain. The fermentation product was purified through staged alcohol precipitation method; and its relative molecular mass was detected by discontinuous SDS-PAGE. The results showed that if removed the precipitate of centrifugation after been treated with 40%（volume fraction） alcohol and then pre- cipitate with 60% alcohol, the freeze-dried sediment was chitosanase with high recovery rate and purity. The relative molecu- lar weight of chitosanase was about 41.7 ku. The optimal reaction temperature and pH for chitosanase was 45 ℃ and 5.5 re- spectively. Chitosanase was stable at 4 ℃ while it would lost activity in very short time above 65 ℃.

关 键 词：壳聚糖酶 发酵 纯化 酶活力 稳定性 

分 类 号：TQ925.9[轻工技术与工程—发酵工程]