Wang Shu-Yang, Bo Yong-Heng, Zhou Xiang, Chen Ji-Hong, Li Wen-Jian, Liang Jian-Ping, Xiao Guo-Qing, Wang Yu-Chen, Liu Jing, Hu Wei, Jiang Bo-Ling
Institute of Modern Physics, Chinese Academy of Sciences, 509 Nanchang Rd., Lanzhou, Gansu 730000, China; Lanzhou University, 222 South Tianshui Road, Lanzhou, Gansu 730000, China.
Institute of Veterinary Drug Quality Inspection of Shandong Province, Jinan 250022, China.
Biomed Res Int. 2017;2017:5373262. doi: 10.1155/2017/5373262. Epub 2017 Jan 24.
Heavy-ion irradiation technology has advantages over traditional methods of mutagenesis. Heavy-ion irradiation improves the mutation rate, broadens the mutation spectrum, and shortens the breeding cycle. However, few data are currently available regarding its effect on morphology and productivity. In this study, the influence of heavy-ion irradiation on when cultivated in approximately 10 L stirred-tank bioreactors was investigated. The specific productivity of the avermectin (AVM) B1a-producing mutant 147-G8 increased notably, from 3885 to 5446 g/mL, approximately 1.6-fold, compared to the original strain. The mycelial morphology of the mutant fermentation processes was microscopically examined. Additionally, protein and metabolite identification was performed by using SDS-PAGE, 2- and 3-dimensional electrophoresis (2DE and 3DE). The results showed that negative regulation gene deletion of mutants led to metabolic process upregulating expression of protein and improving the productivity of an avermectin B1a. The results showed that the heavy-ion beam irradiation dose that corresponded to optimal production was well over the standard dose, at approximately 80 Gy at 220 AMeV (depending on the strain). This study provides reliable data and a feasible method for increasing AVM productivity in industrial processes.
重离子辐照技术相较于传统诱变方法具有优势。重离子辐照提高了突变率,拓宽了突变谱,并缩短了育种周期。然而,目前关于其对形态和生产力影响的数据较少。在本研究中,研究了重离子辐照对在约10升搅拌罐生物反应器中培养时的影响。与原始菌株相比,产生阿维菌素(AVM)B1a的突变体147 - G8的比生产力显著提高,从3885增加到5446克/毫升,约为1.6倍。对突变体发酵过程的菌丝形态进行了显微镜检查。此外,通过使用SDS - PAGE、二维和三维电泳(2DE和3DE)进行蛋白质和代谢物鉴定。结果表明,突变体的负调控基因缺失导致代谢过程中蛋白质表达上调,提高了阿维菌素B1a的生产力。结果表明,对应最佳产量的重离子束辐照剂量远超过标准剂量,在220 AMeV时约为80 Gy(取决于菌株)。本研究为提高工业生产过程中AVM的生产力提供了可靠数据和可行方法。