琼脂柱预筛选法在头孢菌素菌种选育中的应用

Application of Agar Column Pre-screening Method in Breeding of Cephalosporin Strains

[目的]获得高产稳定的头孢菌素优良菌株。[方法]先经UV诱变处理深冷孢子液,再培养单菌落,经富集培养划菌落带,采用琼脂柱预筛选法,通过摇瓶初筛、复筛及斜面传代单菌落菌型考察,研究琼脂柱预筛选法在头孢菌素菌种选育中的应用价值及稳定性。[结果]与单菌落分离直接摇瓶筛选法相比,采用琼脂柱预筛选法筛选单菌落省时、省力,效果好。经琼脂柱预筛选法获得的菌株性能稳定,摇瓶效价平均值高,F1、F2、F3、F4代的摇瓶效价分别高出对照16.7%、33.3%、16.0%、12.0%,其中F2代斜面可确认为生产用斜面。[结论]应用琼脂柱预筛选法进行菌种选育,可提高筛选效率和数量,增加高产菌株的获得几率,缩短筛选时间。

[Objective] The study was to obtain superior strains of cephalosporin with high yield and good stability.[Method] The spore liquid was deeply cooled with UV mutagenic treatment,and then the single colony was cultured,and its colony belt was divided through enrichment culture.With agar column pre-screening method,through the investigation on bacterial type of single colony by primary and secondary screen with shake flask and by slant subculturing,the application value and stability of agar column pre-screening method in breeding of cephalosporin strains were studied.[Result] Compared with direct screening method on single colony separating with shake flask,using agar column pre-screening method to select single colony could get the advantages of time saving,labor saving and good effect.The performance of strains selected by agar column pre-screening method was stable,and the average titer value of shake flask was higher,the titer of shake flask in F1,F2,F3 and F4 were 16.7%,33.3%,16.0% and 12.0% higher than that of CK.and the slant of F2 could be confirmed as the production slant.[Conclusion] When agar column pre-screening method was used to breed strains,the screening efficiency and quantity could be increased,the probability of gaining high yielding strains could be increased and the screening time could be shortened.