微孔板与响应面相结合筛选壳聚糖酶高产菌株

Optimization for screening high chitosanase-producing strain by microplate and Response Surface Methodology

利用微孔板与响应面相结合筛选壳聚糖酶高产茵株。经平板透明圈法从土样中筛选出60株产壳聚糖酶菌株,再经96孔板复筛得到1株产壳聚糖酶活力较高的菌株。通过Plackett-Bunnan设计实验确定蛋白胨、胶体壳聚糖、CaCl_2对产酶具有显著影响;最后应用中心组合设计和响应面分析得到以上3个因素的最佳浓度分别为(%,w/v):蛋白胨1.01%、胶体壳聚糖1.05%、CaCl_2 0.53%。在优化后的培养基中该菌株产壳聚糖酶达到34.95 U/mL,比优化前提高了2.31倍。

Microplate combinded with Response Surface Methodology were used to screen high chitosanase-producing strain. 60 strains producing chitosanase were isolated from soil samples by plate screening, and then one of the higher chitosanase-producing strains was screened by 96 well plates-ELIASA Reader. Plaekett-Burman design was applied to evaluate three important factors affecting chitosanase production, which were colloid chitosan, pepeone and CaCl2. The optimal levels of the main factors were obtained by central composite design and response surface analysis, which were pepeone 1.12%, colloid chitosan 1.26%, and CaC12 0.91%. Under the optimal cultural conditions, the activity of chitosanase was 34.95 U/mL, which was 2.31 times higher than that of the initial strain.