转录组分析棒状链霉菌F613-1高产克拉维酸的分子基础  

Characterization of Clavulanic Acid High-yield Streptomyces clavuligerus Industrial Strain by Transcriptome Analysis

作　　者：覃荣活 付加芳[2] 宗工理[2] 王志宇[2] 曹广祥[1,2] QIN Rong-huo FU Jia-fang ZONG Gong-li WANG Zhi-yu CAOGuang-xiang(School of Medicine and Life Sciences, University of Jinan-Shandong Academy of Medical Sciences, Ji＇ nan 250022 Shandong Medicinal Biotechnology Center, Ji＇ nan 250062)

机构地区：[1]济南大学山东省医学科学院医学与生命科学学院,济南250022 [2]山东省医药生物技术研究中心,济南250062

出　　处：《生物技术通报》2017年第9期235-243,共9页Biotechnology Bulletin

基　　金：山东省自然科学基金项目(ZR2016CP04);山东省医学科学院医药卫生科技创新工程(201604)

摘　　要：旨在探究S.clavuligerus工业菌株高产克拉维酸(CA)的分子基础。采用转录组高通量测序,对比标准菌株和高产CA的工业菌株F613-1之间的基因转录水平差异,结合菌株表型差异进行综合分析。结果表型比较显示,F613-1在固体培养基中生长速度、气生菌丝形态、孢子形成等方面与标准菌株存在差异。液体摇瓶培养中,菌体生长速度无明显变化,但F613-1的糖消耗量显著降低,同时CA产量比ATCC27064提高约11.7倍。转录组分析显示,F613-1中CA基因簇及其途径特异性调控基因的表达量显著升高,其副产物全霉素基因簇的表达量显著下降;CA合成原料精氨酸和3-磷酸甘油醛代谢相关基因的转录水平发生显著变化,精氨酸合成代谢基因上调,而3-磷酸甘油醛分解代谢基因下调;ABC转运系统中80多个基因的转录水平也发生明显变化,其中42个显著上调,38个显著下调。工业生产菌株高产克拉维酸的主要原因是CA基因簇及其调控基因表达量的升高、相关副产物基因簇的沉默及CA合成原料精氨酸和3-磷酸甘油醛的积累。This work aims to explore the molecular mechanism of clavulanic acid（CA）high-yield Streptomyces clavuligerus industrialstrain. Transcriptome high-throughput sequencing was performed to compare the differential gene expressions between standard strainATCC27064 and high-yield industrial strain F613-1,and comprehensive analysis was carried out combined with the phenotypic differencesbetween them. The phenotypic results showed that F613-1 grew differently in growth rate,aerial mycelium morphology and spore formation insolid medium,compared with the standard strain. In the liquid culture,the growth rate of the two strains demonstrated no difference,but thesugar consumption of F613-1 significantly decreased,and the CA yield was 11.7 times higher than that of ATCC27064.Transcriptome analysisindicated that the expressions of both CA cluster genes and pathway-specific regulatory genes significantly increased;while the expressionsof holomycin（CA by-product）cluster genes significantly decreased. On the other hand,the transcriptional levels of genes associated witharginine and glyceradldehyde-3P（raw materials for CA synthesis）metabolic pathways changed significantly,i.e.,up-regulated for arginineand down-regulated for glyceradldehyde-3P. Besides,the expressions of over80 genes involved in ABC transport system obviously changed,and 42 remarkably up-regulated and 38 down-regulated. The main reason of F613-1 highly produces clavulanic acid possibly lies in theincreased transcription of CA cluster genes and relative regulatory genes,the silence of cluster genes involved in CA by-products,and theaccumulation of arginine and alyceradldehyde-3P.

关 键 词：棒状链霉菌 工业菌株 转录组 克拉维酸 

分 类 号：Q933[生物学—微生物学]