用原生质体融合技术选育谷氨酸高产菌

BREEDING OF EASY-EXTRACTED HIGH PRODUCING GLUTAMATE STRAIN USING THE PROTOPLAST FUSION TECHNIQUE

利用原生质体融合技术,在不对亲株做任何诱变的前提下,成功地使天津短杆菌TG-866与钝齿棒杆菌B9的原生质体进行融合,获得兼有两亲株遗传性状——细胞个体大、产酸高的融合子F263及F288。确定了两亲株原生质体制备、再生及融合的最佳条件。在该条件下,其原生质体形成率均在99.9%以上,再生率分别为23%及28%,原生质体融合率为3.2×10^(-3)。在最佳摇瓶发酵条件下,F263及F288的谷氨酸产率分别为8.4g/dl及8.03g/dl。通过连续10次摇瓶传代,发现该融合子较稳定,因而具有较高的应用价值。

Using the protoplasts fusion technique, Brevibacterium tianjinese TG-866 and Corynebacterium crenatum B9 were successfully fused. Fusion hybrids F288 and F263 were obtained which contained hereditary properties of the parents i.e. big size and high acid productivity.The best condition for protoplast formation, regeneration and fusion were found. Under these conditions, the rate of obtained protoplast was more than 99.9%, the rate of regeneration was respectively 23% and 28%, the rate of protoplast's fusion was 3.2×10^(-(?)). The best condition of flask-shaking fermentation of F263 and F288 was found. The producing of glutamic acid was respectively 8.4 g/dl and 8.02 g/dl. after 10 generations in the shaking-flask, they keep the original characteristics and have high applied value.