摘要
偶发分枝杆菌(Mycobacterium fortuitum)作为重要的9α-羟基雄甾-4-烯-3,17-二酮(9α-OH-AD)生产菌株,在甾体药物的生产中起着重要的作用。为了进一步提高偶发分枝杆菌的9α-OH-AD生产水平,对原始菌株进行常压室温等离子体(ARTP)诱变处理。在最佳诱变处理时间(35 s)下,筛选得到一株高产9α-OH-AD的诱变株Mycobacterium fortuitum AR-2。当投加10 g/L植物甾醇时,转化96 h,该菌株9α-OH-AD摩尔生成率达87.6%,较原始菌株提高14.9%。进一步,对AR-2进行ARTP-LiCl复合诱变,确定最佳LiCl诱变浓度为3.5%,得到一株高产9α-OH-AD的诱变菌株Mycobacterium fortuitum#91。当RM-β-CD添加量与底物摩尔比为1:1,投加10 g/L植物甾醇时,该菌株9α-OH-AD摩尔生成率达95.7%,较原始菌株提高34.9%,具有良好的应用前景。
Mycobacterium fortuitum is an important strain for the production of 9α-hydroxyandrost-4-ene-3,17-dione(9α-OH-AD)in the manufacture of steroid drugs.To improve the 9α-OH-AD production of Mycobacterium fortuitum,the original strain was subjected to atmospheric pressure room temperature plasma(ARTP)mutagenesis treatment.A mutant strain,Mycobacterium fortuitum AR-2,with high 9α-OH-AD production was screened at the optimal mutagenic treatment time(35 s).When transformed for 96 h with 10 g/L phytosterol,the strain achieved a molar production rate of 9α-OH-AD of 87.6%,which was 14.9%higher than that of the original strain.Furthermore,AR-2 was subjected to ARTP-LiCl compound mutagenesis,the optimal LiCl mutagenesis concentration was determined to be 3.5%,and a mutagenic strain Mycobacterium fortuitum#91 with high 9α-OH-AD production was obtained.When the molar ratio of RM-β-CD addition to phytosterol was 1:1 and 10 g/L phytosterol was administered,the molar production rate of 9α-OH-AD of this strain reached 95.7%,which was 34.9%higher than that of the original strain,and it has a good application prospect.
作者
高华义
杨佳浩
何蔓
苏振华
周海杰
申雁冰
王敏
GAO Hua-yi;YANG Jia-hao;HE Man;SU Zhen-hua;ZHOU Hai-jie;SHEN Yan-bing;WANG Min(Key Laboratory of Industrial Fermentation Microbiology,Ministry of Education,Tianjin Key Laboratory of Industrial Microbiology,College of Biotechnology/Tianjin University of Science&Technology,Tianjin 300457,China)
出处
《山东农业大学学报（自然科学版）》
北大核心
2023年第5期657-662,共6页
Journal of Shandong Agricultural University：Natural Science Edition
基金
国家自然科学基金(21978221)。
关键词
偶发分枝杆菌
诱变选育
诱变菌
Mycobacterium fortuitum
mutation breeding
mutagenesis
作者简介
第一作者:高华义(1989-),男,博士研究生,专业方向:发酵工程.E-mail:gaohuayi2013@163.com;通讯作者:申雁冰.E-mail:shenyb@tust.edu.cn;通讯作者:王敏.E-mail:minw@tust.edu.cn。
