摘要
以紫外(UV)与亚硝基胍诱变的竹黄菌(Shiraia bambusicola)菌株NU12和UV4为出发菌株,60℃处理5 min、紫外(距离30 cm,30 W)照射90 s进行双亲原生质体灭活,通过30%聚乙二醇PEG6000介导原生质体融合20 min。结合平板初筛和高效液相色谱(HPLC)分析进行复筛,通过3轮重组融合操作,筛选出6株高产竹红菌甲素的融合株。其中融合菌株FIII-21的竹红菌甲素产量达到80.4 mg/L,比原始出发菌株提高了58.9%～167.1%,且遗传稳定,具有较高的医药及工业应用价值。
Two mutant strains,i.e.,NU12 and UV4,of Shiraia bambusicola,treated by UV and nitrosoguanidine mutagenesis were breeding candidates for higher production of hypocrellin A by genome-shuffling.The protoplasts were inactivated by heat at 60 ℃ for 5 min and UV treatment for 90 s,respectively.The inactive protoplasts were fused and regenerated in the presence of polyethylene glycol（PEG6000,30%,W/V） for 20 min.The selective plates with hypocrellin pigment for observation and the assay of high performance liquid chromatography were used to screen the mutant strain of higher production of hypocrellin A.After two rounds of genome-shuffling,six strains of mutants with higher yield were obtained.The production of hypocrellin A by mutant FIII-21 reached 80.4 mg/L,and it increased by 58.9%-167.1% than that of the original strains.The production of the mutant was verified to be genetically stable.
出处
《生物加工过程》
CAS
CSCD
2012年第1期25-29,共5页
Chinese Journal of Bioprocess Engineering
基金
教育部留学基金资助项目(K513201011)
国家大学生创新性实验计划资助项目(5731502710)
关键词
竹红菌甲素
原生质体融合
竹黄菌
hypocrellin A
protoplast fusion
Shiraia bambusicola
