克拉维酸高产菌的诱变育种及发酵工艺研究  

Mutagenesis breeding of clavulanic acid producing strain and optimization of fermentative conditions

作　　者：宗工理[1,2] 钟传青[3] 王新圆 覃荣活 曹广祥[1,2] 

机构地区：[1]山东省医药生物技术研究中心,济南250062 [2]济南大学山东省医学科学院医学与生命科学学院,济南250200 [3]山东建筑大学市政与环境工程学院,济南250101

出　　处：《中国抗生素杂志》2017年第2期112-116,共5页Chinese Journal of Antibiotics

基　　金：山东省自然科学基金培养基金项目(No.ZR2016CP04);山东省医学科学院医药卫生科技创新工程(No.201604)

摘　　要：目的为满足从棒状链霉菌突变库中筛选棒酸高产菌株的需求,建立一种新的克拉维酸高产菌株筛选方法。方法以克拉维酸生产菌株—棒状链霉菌(Streptomyces clavuligerus)SC51为出发菌株,采用紫外-NTG联合诱变方法,以链霉素抗性作为筛选标记,从不断增加链霉素浓度的平板上,筛选链霉素抗性突变菌株,以此突变菌株为实验菌株,对碳源、氮源、前体三油酸甘油酯和发酵条件进行优化。结果在链霉菌浓度60μg/m L的平板上筛选到链霉素抗性突变菌株SCM0943,效价达到2955μg/m L,且遗传性能稳定;优化的发酵条件下,摇瓶发酵120h,克拉维酸效价可达3437μg/m L,较出发菌株提高49.1%。结论棒状链霉菌的链霉素耐受性与克拉维酸的产量存在正相关性,本试验结果对克拉维酸工业化生产具有很好的参考价值。Objective To facilitate the screening of a large number of Streptomyces clavuligerus mutants, we developed a screening method. Methods The clavulanic acid-producing strain Streptomyces clavuligerus SC51 was treated by UV irradiation and NTG approaches. The streptomycin tolerant mutants were selected on the agar plate containing streptomycin. Mutant strain with the highest yield of clavulanic acid（CA）was used as the parent strain, and the fermentation medium （carbon sources, nitrogen sources, triglyceride） and condition （initial pH, inoculation amount and liquid volume in flask） were optimized. Results The mutant strain, named SCM0943, was selected from the agar plate, which contains 60μg/mL streptomycin and the productivity of CA reached 2955μg/mL. Under optimized conditions, the maximum potency of 3437μg/mL was obtained afterl20h-fermentation in flask, and the maximum potency increased 1.49 times compared with the original strain. Conclusion Our resultsindicated a positive correlation between the tolerance of streptomycin and the clavulanic acid yield, and provideda reference for the commercial productionof CA.

关 键 词：克拉维酸 诱变育种 链霉素筛选 突变株 

分 类 号：R978.1[医药卫生—药品]