摘要
新霉素主要是由弗氏链霉菌(Streptomyces fradiae)发酵生产的一种氨基糖苷类抗生素,广泛应用于医药、畜牧等领域,能够有效地控制和治疗细菌引起的疾病,提高畜禽的生长速度和生产性能。为了提高新霉素产量,该研究通过比较转录组学分析原始弗氏链霉菌SF-1和诱变高产菌SF-2的转录组数据,挖掘出转录水平差异显著的转录因子,然后过表达构建重组质粒,再通过接合转移构建出工程菌株。经摇瓶发酵验证筛选出对新霉素合成影响显著的工程菌株,进行启动子优化最终得到高产的生产新霉素工程菌株SF-2/pP-NeyR。随后对发酵培养基组分进行正交优化,最终新霉素B效价为11843 U/mL。综合优化后新霉素B的效价较初始提高了29.6%。该实验结果为新霉素后续代谢机理研究、其和弗氏链霉菌新霉素合成基因簇的相互作用研究、解析调控新霉素合成机理奠定了基础,为进一步提高新霉素效价提供了思路。
Neomycin is an aminoglycoside antibiotic mainly produced by Streptomyces fradiae.It is widely used in medicine,animal husbandry and other fields.It can effectively control and treat diseases caused by bacteria,and improve the growth speed and production performance of livestock and poultry.To improve the production of neomycin,the transcriptome data of the original Streptomyces fradiae SF-1 and the mutagenic high-yield strain SF-2 were analyzed by comparative transcriptomics,and the transcription factors with significant differences in transcription levels were mined,and then the recombinant plasmids were constructed by overexpression,and then the engineered strains were constructed by conjugation transfer.The engineering strain with significant effect on neomycin synthesis was screened by shake flask fermentation,and the high-yield neomycin producing engineering strain SF-2/pP-NeyR was finally obtained by promoter optimization.Then the components of fermentation medium were orthogonal optimized,and the final titer of neomycin B was 11843 U/mL.After comprehensive optimization,the titer of neomycin B increased by 29.6%compared with the initial one.The experimental results laid a foundation for the follow-up study of metabolic mechanism,the study of its interaction with Streptomyces fradiae neomycin synthesis gene cluster,the analysis and regulation of neomycin synthesis mechanism,and provided ideas for further improving the potency of neomycin.
作者
程一晗
李翔飞
韩如梦
刘艳
薛正莲
CHENG Yihan;LI Xiangfei;HAN Rumeng;LIU Yan;XUE Zhenglian(College of Bio-logic&Food Engineering,Anhui Polytechnic University,241000,China;Engineering Laboratory for Industrial Microbiology Molecular Beeding of Anhui Province,241000,China)
出处
《食品与发酵工业》
CAS
CSCD
北大核心
2024年第24期84-90,共7页
Food and Fermentation Industries
基金
国家自然科学基金项目(32300059)
安徽省博士后研究人员科研活动经费资助项目(2023B692)。
关键词
弗氏链霉菌
新霉素
比较转录组学
转录因子
启动子优化
发酵优化
Streptomyces fradiae
neomycin
comparative transcriptomics
transcription factors
promoter optimization
fermentation optimization
