复合诱变选育氢化可的松高转化率菌株Breeding-山东农业大学.PDF

山东农业大学学报 自然科学版( ),2018,49(3):396-401 VOL.49NO.32018 Journal of ShandongAgriculturalUniversity (Natural ScienceEdition ) doi:10.3969/j.issn.1000-2324.2018.03.006 复合诱变选育氢化可的松高转化率菌株 * * 薛玮莹申雁冰 黄 炜王 敏 邓铭倩, , , , 工业发酵微生物教育部重点实验室, 天津市工业微生物重点实验室, 天津科技大学生物工程学院, 天津 300457 摘 要: 蓝色犁头霉 （Absidiacoerulea）是转化17α-羟基孕甾-4-烯-3,20-二酮-21-醋酸酯 （RSA）生成氢化可的松 （简 称HC）的生产菌株。因保藏菌种退化，为了进一步提高HC 生产水平，对保藏菌株分离纯化后，利用常压室温等离 子体诱变 （简称ARTP）和ARTP-LiCl 复合诱变的方法对一株蓝色犁头霉进行了诱变。建立了以菌落形态为标准的 初筛方法，选取培养6d 后菌落颜色较深、背面呈环状纹路、菌丝较密集的菌株进行HC 转化复筛，得到一株遗传稳 定性较好的菌株AL-172，RSA 浓度为3.5g/L 时，HC 转化率达72.52%，底物投料浓度和HC 转化率较实验室保藏菌 40% 16.20% 株分别提高了 和 ，具有良好的应用前景。 关键词: 氢化可的松; 蓝色犁头霉; 诱变育种; 菌落形态;ARTP 中图法分类号:Q93-335 文献标识码:A 文章编号:1000-2324(2018)03-0396-06 Breeding Strains with Compound Mutation for High Conversion RateofHydrocortisone * * XUEWei-ying,SHENYan-bing ,HUANGWei,WANGMin ,DENGMing-qian Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, Collegeof Biotechnology/Tianjin University of Science Technology, Tianjin300457, China Abstract: Absidia coerulea is a production strain that converts 17 alpha-hydroxyprogesterone - 4-ene-3, 20-di-21-acetate (RSA) into hydrocortisone (HC). The preserved strain could degenerate, in order to further improve the level of HC production, after the isolation and purification of the preserved strain, the mutagenesis was carried out by using the compound mutagenesis method of Atmospheric temperature Plasma (art) and ARTP-LiCl. The primary screening method was established on the basis of colony morphology. After 6 days of culture, the colony color was darker, there was cricoid markings on the back, HC conver