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2025年8月3日 星期日
农业生物技术学报  2019, Vol. 27 Issue (7): 1322-1330    DOI: 10.3969/j.issn.1674-7968.2019.07.020
  研究资源与技术改进 本期目录 | 过刊浏览 | 高级检索 |
核糖体工程技术选育阿维拉霉素高产菌株
刘华华, 陈宇航, 陈敏*
浙江工商大学 食品与生物工程学院,杭州 310018
Breeding of High Avilamycin-producing Strains by Ribosome Engineering
LIU Hua-Hua, CHEN Yu-Hang, CHEN Min*
School of Food Sciences and Biotechnology, Zhejiang Gongshang University, Hangzhou 310018, China
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摘要 阿维拉霉素是一种新型的代谢调节剂和消化促进剂,被广泛应用于国内外畜禽养殖中。菌种产素能力低、发酵工艺不成熟等因素限制了阿维拉霉素在国内的工业化生产。本研究通过核糖体工程技术,采用链霉素对实验室保藏的绿色产色链霉菌菌株(Streptomyces viridochromogenes gs 77)进行抗性选育,以获得阿维拉霉素高产突变菌株。经过药敏试纸法和高效液相色谱法(HPLC)筛选获得一株阿维拉霉素高产突变菌株S. viridochromogenes gs 77-54,摇瓶第8天时阿维拉霉素产量达到最大,为出发菌株的1.80倍,传代实验表明突变菌株高产性能遗传稳定。对突变前后菌株的形态特征和rpsl基因(编码核糖体蛋白S12)分析,结果显示突变菌株的气生菌丝长直形、较为粗壮,孢子为直径约1.1 µm×0.6 µm的椭圆形,与出发菌株存在明显区别。rpsl基因序列分析结果发现,该基因片段出现点突变,第356位的G突变为A,由精氨酸突变为谷氨酰胺。本研究筛选到阿维拉霉素高产突变菌株,为绿色产色链霉菌菌种选育提供了新的参考资料。
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刘华华
陈宇航
陈敏
关键词 阿维拉霉素核糖体工程菌株选育链霉素抗性    
Abstract:Avilamycin is widely used as a new type of metabolic regulator and digestive enhancer in livestock feeding. The low production capacity of the strain and the immature fermentation process limit the industrial of avilamycin in China. Ribosome engineering technique was used to carry out resistance breeding of the Streptomyces viridochromogenes gs 77 by streptomycin to obtain a high-yield mutant strain of avilamycin, . Through the drug sensitive test and high performance liquid chromatography, a high-yield avilamycin mutant S.viridochromogenes gs 77-54 was obtained. The yield of avilamycin reached the maximum at 8th day of shake flask, and increased to 1.80 fold than that of the original strain. Passage experiments showed that the high-yield performance of mutant strains was genetically stable. rpsl gene (encoding the ribosomal protein S12) and the morphological characteristics of the strains parent and after the mutation were analyzed. The results showed that the aerial hyphae of the mutant strain were long and straight, and the spores were elliptical with a diameter of about 1.1 µm× 0.6 µm, which was significantly different from the starting strain. Sequence analysis of the rpsl gene revealed a point mutation in the gene fragment, the G mutation at position 356 was mutated A, which was mutated to glutamine by arginine. The avilamycin high-yield mutant strain was constructed successfully, which would provide a new idea for the breeding of S. viridochromogenes.
Key wordsAvilamycin    Ribosome engineering    Breeding of strains    Streptomycin resistance
收稿日期: 2018-11-26     
ZTFLH:  S182  
  Q815  
基金资助:浙江省基础公益研究计划项目(No. LGN18C010001)
通讯作者: chenmin@zjgsu.edu.cn   
引用本文:   
刘华华, 陈宇航, 陈敏. 核糖体工程技术选育阿维拉霉素高产菌株[J]. 农业生物技术学报, 2019, 27(7): 1322-1330.
LIU Hua-Hua, CHEN Yu-Hang, CHEN Min. Breeding of High Avilamycin-producing Strains by Ribosome Engineering. 农业生物技术学报, 2019, 27(7): 1322-1330.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2019.07.020     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2019/V27/I7/1322
 
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