他克莫司高产菌株选育及发酵配方优化

Breeding of tacrolimus high-yield strain and optimization of fermentation conditions

目的 选育他克莫司高产菌株,优化培养基配方,提高发酵产量。方法 采用原生质体融合技术对2株筑波链霉菌进行基因组重排,并经响应面设计对发酵培养基配方进行优化。结果 以紫外灭活作为遗传标记经过4轮基因组重排育种后,摇瓶筛选获得3株高产菌株,其中菌株FK22-71菌丝球松散、椭圆状,发酵浸泡液颗粒状。该菌株稳定高产,采用响应面设计优化后的配方在50 L罐发酵产量平均达1684 mg/L。结论 基因组重排技术应用于他克莫司高产菌株选育,可大幅提高发酵产量,为其工业化发酵生产奠定了基础。

Objective Breeding high-yield strains of tacrolimus and optimizing the culture medium formula to improve the fermentation unit.Methods The protoplast fusion technique was used to rearrange the genome of two Streptomyces tsuknbaenis strains,and the fermentation medium was optimized by response surface design.Results After four rounds of genome shuffling breeding with UV inactivation as genetic marker,three high-yield strains were obtained by shaking flask screening.Among them,the mycelium ball of strain FK22-71 was loose and oval,and the fermentation soaking liquid was granular.In 50 L fermentor,the average production of tacrolimus reached 1684 mg/L under the response surface optimization by the strain FK22-71,which was steady and high-yielding.Conclusion The application of genome shuffling technology in the breeding of high-yielding tacrolimus producing strains can significantly increase production and lay the foundation for industrial fermentation production.