维吉尼亚链霉菌物理诱变及外源添加氯化镧对VGM产量的影响  

Studies on the physical mutagenesis of virginiamycin producing Streptomyces virginiae and the impact of lanthanum on antibiotic productivity

作　　者：周跃慧 仝倩倩 袁丽霞 陈祥松 孙立洁 吴金勇 姚建铭[1] Zhou Yue-hui;Tong Qian-qian;Yuan Li-xia;Chen Xiang-song;Sun Li-jie;Wu Jin-yong;Yao Jian-ming(Institute of Plasma Physics Chinese Academy of Sciences,Hefei 230031;University of Science and Technology of China,Hefei 230026)

机构地区：[1]中国科学院合肥物质科学研究院等离子体物理研究所,合肥230031 [2]中国科学技术大学,合肥230026

出　　处：《中国抗生素杂志》2018年第11期1389-1396,共8页Chinese Journal of Antibiotics

基　　金：国家自然科学基金青年科学基金项目(No.21506235);安徽省自然科学基金面上项目(No.1508085MB38);中科院等离子体物理研究所科学基金(No.DSJJ-15-YY02);中科院等离子体物理研究所科学基金(No.DSJJ-16-YY02)

摘　　要：目的利用物理诱变结合链霉素抗性筛选维吉尼亚霉素(VGM)高产菌株维吉尼亚链霉菌,并考察外源添加稀土元素氯化镧对维吉尼亚链霉菌发酵的影响。方法一方面,以链霉素为筛选剂建立孢子致死突变标志的微波及紫外诱变两种筛选模型,采用管碟法获得抑菌圈直径较大的作为初筛菌株,再通过摇瓶复筛进一步确定高产菌株。另一方面,在发酵培养基中外源添加3～30mg/L的氯化镧,考查其对链霉菌合成VGM的影响。结果比较两种筛选模型,基于链霉素的微波诱变效果较好,正突变率高达32.83%;并获得一株高产菌株MW 178(146.72±11.80)μg/mL,较出发菌株(21.24±1.34)μg/mL提高了约6倍。在发酵初始时,外源添加10mg/L氯化镧,VGM的产量由146.72μg/mL提高到180.01μg/mL,提高了约22.69%。结论采用微波诱变结合链霉素抗性筛选的方法可以获得VGM高产突变株;在发酵初始时,外源添加适量氯化镧可以显著提高VGM产量。Objective A method of physical mutagenesis coupled with streptomycin resitance screening was applied to select virginiamycin （VGM） high-producing strain. Also, the effect of adding lanthanum chloride on the production of VGM was studied. Methods First, two streptomycin-resistance mutational labelling selection models with physical mutagenic factor （microwave and ultraviolet） were built to screen high-yield mutants. Then preliminary positive strains were picked out by measurement of the inhibitory activity to Micrococcus luteus. And the subsequent determination by HPLC were carried out for the high VGM-producing strains cultivated in the shake-flask. Finally, lanthanum chloride was added in fermentation medium of the strain with the concentration of 3-30mg/L. Results The mutagenesis of microwave combined with streptomycin resistance was better and the positive mutation rate was up to 32.83%. Streptomycin-resistant mutant strain MW 178 was got by microwave producing VGM with yield of （146.72±11.80）μg/mL which beyond six times higher than the initial strain （21.24±1.34）μg/mL. The production of MW 178 reached up to 180.01μg/mL eventually by adding the concentration of 10mg/L lanthanum chloride to the medium at the beginning of the fermentation. Conclusion Microwave mutation compound streptomycin resistance screening is effetive for screening high-yeilding mutants and the addition of lanthanum chloride could significantly improve the yield of VGM.

关 键 词：维吉尼亚链霉菌 维吉尼亚霉素 氯化镧 选育 微波及紫外诱变 

分 类 号：Q939[生物学—微生物学]