转录组分析棒状链霉菌F613-1高产克拉维酸的分子基础

doi:10.13560/j.cnki.biotech.bull.1985.2017-0275

摘要/Abstract

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。

关键词: 棒状链霉菌, 工业菌株, 转录组, 克拉维酸

Abstract: This work aims to explore the molecular mechanism of clavulanic acid（CA）high-yield Streptomyces clavuligerus industrial strain. Transcriptome high-throughput sequencing was performed to compare the differential gene expressions between standard strain ATCC27064 and high-yield industrial strain F613-1,and comprehensive analysis was carried out combined with the phenotypic differences between them. The phenotypic results showed that F613-1 grew differently in growth rate,aerial mycelium morphology and spore formation in solid medium,compared with the standard strain. In the liquid culture,the growth rate of the two strains demonstrated no difference,but the sugar consumption of F613-1 significantly decreased,and the CA yield was 11.7 times higher than that of ATCC27064.Transcriptome analysis indicated that the expressions of both CA cluster genes and pathway-specific regulatory genes significantly increased；while the expressions of holomycin（CA by-product）cluster genes significantly decreased. On the other hand,the transcriptional levels of genes associated with arginine and glyceradldehyde-3P（raw materials for CA synthesis）metabolic pathways changed significantly,i.e.,up-regulated for arginine and 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 the increased transcription of CA cluster genes and relative regulatory genes,the silence of cluster genes involved in CA by-products,and the accumulation of arginine and alyceradldehyde-3P.

Key words: Streptomyces clavuligerus, industrial strain, transcriptome, clavulanic acid

覃荣活,付加芳,宗工理,王志宇,曹广祥,. 转录组分析棒状链霉菌F613-1高产克拉维酸的分子基础[J]. 生物技术通报, 2017, 33(9): 235-243.

QIN Rong-huo,FU Jia-fang,ZONG Gong-li,WANG Zhi-yu,CAO Guang-xiang,. Characterization of Clavulanic Acid High-yield Streptomyces clavuligerus Industrial Strain by Transcriptome Analysis[J]. Biotechnology Bulletin, 2017, 33(9): 235-243.

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