常温常压等离子体诱变选育安普霉素高产菌株  

Breeding of High-yielding Strains of Apramycin by Atmospheric and Room-temperature Plasma

作　　者：张鄂 王海波 王洛菊 李敏 孟敏 田伟波 赵国毅 李继安[2] ZHANG E;WANG Hai-bo;WANG Luo-ju;Li Min;MENG Min;TIAN Wei-bo;ZHAO Guo-yi;LI Ji-an(Shandong Qilu King-Phar Pharmaceutical CO.,LTD.,Jinan 250400,China;State Pharmaceutical Administration of Shanghai Institute of Pharmaceutical Industry,Shanghai 200000,China)

机构地区：[1]山东齐发药业有限公司,济南250400 [2]上海医药工业研究院有限公司,上海200000

出　　处：《中国兽药杂志》2023年第4期34-39,共6页Chinese Journal of Veterinary Drug

摘　　要：安普霉素作为一种广谱兽用抗生素,对多种革兰氏阴性菌、阳性菌和某些支原体具有较好的抗菌作用。本论文利用常温常压等离子体(ARTP)技术结合抗性筛选方法,对安普霉素产生菌黑暗链霉菌进行诱变选育,以期得到高产菌株。以AP-520为出发菌,进行ARTP诱变处理,ARTP最佳诱变时间是70 s,处理后的菌液在潮霉素平板上进行抗性筛选,单菌落经过48孔板大量筛选后再进行摇瓶初筛和复筛,共进行8轮筛选试验,得到一株高产菌株AP-6023,摇瓶效价达到11444 U·ml-1,较对照提高29%,经过10次传代培养后仍能保持产抗能力,30 L发酵罐效价为10920 U·ml-1,较对照提高23%。突变株AP-6023的获得,大大提高了安普霉素的生产水平,证实了ARTP诱变技术的有效性,试验方法为其它工业微生物的育种提供了解决思路。As a broad-spectrum veterinary antibiotic,apramycin has a good antibacterial effect on a variety of Gram-negative bacteria,positive bacteria and some mycoplasmas.In order to obtain high yield strains,we used the technology of atmospheric and room-temperature plasma(ARTP)combined with resistance screening method,to mutagenesis and breeding of apramycin producing bacteria streptomyces tenebrariu.AP-520 was used as the starting bacterium for ARTP mutagenesis.The optimal mutagenesis time of ARTP was 70 s.The treated bacterial solution was screened for resistance on hygromycin plate.After a large number of single colonies were screened by 48-well plates and then subjected to primary screening and re-screening in shake flasks.A total of 8 rounds of screening tests were conducted,and a high-yield strain AP-6023 was obtained.The titer of shaking flask reached 11444 U·ml-1,which was 29%higher than that of the control.After 10 subcultures,AP-6023 could still maintain the ability to produce antibiotics,and the titer of the 30 L fermenter was 10920 U·ml-1,which was 23% higher than the control.The acquisition of mutant strain AP-6023 greatly improved the production level of apramycin,and confirmed the effectiveness of ARTP mutagenesis technology.The test method provides a solution for the breeding of other industrial microorganisms.

关 键 词：安普霉素 常温常压等离子体(ARTP) 抗性筛选 高产菌株 

分 类 号：S859.79[农业科学—临床兽医学]