中国农业科学 ›› 2007, Vol. 40 ›› Issue (7): 1416-1421 .

• 植物保护 • 上一篇    下一篇

原生质体融合技术对秦岭霉素产量提高的影响

龙建友,唐世荣,吴文君   

  1. 广州大学环境科学与工程学院
  • 收稿日期:2006-01-25 修回日期:1900-01-01 出版日期:2007-07-10 发布日期:2007-07-10
  • 通讯作者: 吴文君

Novel antibiotics——Studies on bioactivity of qinlingmycin and protoplast fusion of its producing strain

  1. 广州大学环境科学与工程学院
  • Received:2006-01-25 Revised:1900-01-01 Online:2007-07-10 Published:2007-07-10

摘要: 【目的】对No.24菌株及其诱变株Ms-24菌株的原生质体的制备、融合、再生条件及融合子的生物活性进行了研究,探索了秦岭霉素的产量提高与融合条件的关系。【方法】采用荧光染色技术对融合子进行了标记,秦岭霉素含量的测定采用了HPLC法进行。【结果】融合后得到的3株高产秦岭霉素的融合子即为pf77,pf126和pf138菌株,其发酵产物的杀菌活性比原始No.24菌株分别提高了43.48%,60.87%和65.22%。HPLC检测发现,融合子pf77,pf126,pf138菌株发酵产物中秦岭霉素的含量与No.24菌株相比分别提高了36.59%,74.39%和91.46%。【结论】进行原生质体再生时,添加甘氨酸、溶菌酶的浓度要适中;原生质体融合时,酶解时间、聚乙二醇(PEG)的浓度和分子量也要适宜。

关键词: 秦岭霉素, 原生质体融合, 融合子

Abstract: a novel lactam antibiotic, qinlingmycin, was isolated from fermentation products of one Streptomyces separated from the soil of qingling mountain. The bioactivity of qinlingmycin had been compared with Penicillin and Cephradine. the result showed that the inhibition activity of qinlingmycin against Bacillus cereus,Bacillus cirulans,Pleudomona aereuginosa were better than Penicillin and Cephradine. and the MIC was 1.2ug/ml,1.5 ug/ml and 2.0ug/ml respectively. against Sarcina Lutea Schroeter,Zscherichia coli was lower than Penicillin and Cephradine. The activity against Staphylococcus aureus is higher than Penicillin lower than Cephradine.No.24 and Ms-24 strain produce qinlingmycin but also pigment.No.24 and mutagensis Ms-24 strain’s protoplast were obtained in lysozyme.the protoplast were mixed at ratio of 1:1 with PEG was added to fuse protoplast.and marking the fusants with fluorescence.. qinlingmycin was obtained through isolation and purification.some relative conditions for preparation, fusion and regeneration of two strain’s protoplast were studied. it was proved that 0.25% Gly was optimal condition for preparation and regeneration of the protoplast. the lysozyme concentration was 1mg/ml,and the optimal treating time was 1h.PEG in different molecular weight tested in the experiment had little influence on fusion of the protoplasts. but its concentration was of importancel.PEG at a concentration on 50% was of benefit to the protoplast fusion. .among 163 fusants, strain, three highly qinlingmycin-producing strain pf77,pf126 and pf138 was obtained. the bioassay results showed that the fungicidal activity of three fusants increased by 43.48%,60.87% and 65.22%。and the production of qinlingmycin increased by 36.59%,74.39% and 91.46%. and three fusants was Successively cultured ten times in the Medium, heredity character was stable.