摘要: |
【目的】从自然环境中筛选、鉴定高效降解羽毛细菌,并通过转录组学筛选该菌株中差异表达基因(DEGs),以探明其降解羽毛的分子机制。【方法】利用梅花桩采样法采集长期堆放羽毛的土壤,利用羽毛发酵培养基筛选分离可高效降解羽毛的细菌,对其进行形态学描述和分子生物学鉴定。以筛选菌株为研究对象,将其分别接种于羽毛发酵培养基(试验组)和LB培养基(对照组)中,培养36 h后收集2个处理组菌体进行转录组测序,之后对试验组与对照组中DEGs进行筛选,利用GO功能注释和KEGG富集分析对DEGs进行注释,探索DEGs在羽毛降解菌降解过程中起关键作用的功能和通路。【结果】筛选得到1株高效降解羽毛的菌株XS6,培养24 h后其能将羽毛发酵培养基(羽毛10.0 g/L)中羽枝降解97%,角蛋白酶活性为201.2 U/mL;且培养36 h后,在培养基中检测到了亚硝酸盐。经形态学和分子生物学鉴定,菌株XS6为白色芽孢杆菌。转录组测序结果表明,在2个处理组之间共筛选出655个DEGs,其中表达上调基因148个,表达下调基因507个。GO功能注释结果显示,DEGs注释到3大类32个功能组中,其中与代谢过程、细胞过程、细胞、细胞部分、催化活性有关的基因注释较多。KEGG富集分析结果显示,DEGs主要在能量和物质代谢途径、信号传导途径、生物合成途径富集。【结论】成功获得了1株高效降解羽毛的白色芽孢杆菌XS6,其在降解羽毛过程中参与了菌株间的协同作用、能量代谢以及生物大分子物质的合成。 |
关键词: 羽毛降解 降解菌 转录组学 降解机制 |
DOI:DOI:10.13207/j.cnki.jnwafu.2024.12.003 |
分类号: |
基金项目:国家科技部专项“国际间重点联合项目”(2017YFE0129900);河南省重点攻关项目(212102110157);河南省重点研发与推广专项(科技攻关)(212102110174) |
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Screening identification and transcriptome analysis of feather-degrading bacterium |
ZHANG Pengzhen1, XUE Linlin1, YANG Xiaojin2, FAN Shouwu2, LI Wang1
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1.College of Animal Science and Technology,Henan University of Science and Technology,Luoyang,Henan 471023,China;2.Luoyang OKBK Biotechnology Co.,Ltd,Luoyang,Henan 471600,China
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Abstract: |
【Objective】This study screened and identified efficient feather-degrading bacteria from natural environment,as well as selected differentially expressed genes(DEGs) in the strain through transcriptomics to explore its molecular mechanism of feather degradation.【Method】Soils containing long-term accumulated feathers were collected using the plum stake sampling method.Feather fermentation medium was used to screen and isolate bacteria capable of efficiently degrading feathers.The isolated bacteria were morphologically described and identified using molecular biology techniques.The selected bacterial strains were activated and then inoculated separately into the feather fermentation medium (experimental group) and LB medium (control group).After 36 hours of cultivation,bacterial cells from both treatment groups were collected for transcriptomic sequencing.DEGs between experimental and control groups were then identified.GO functional annotation and KEGG enrichment analysis were performed to explore the functions and pathway roles of DEGs in biological processes.【Result】A highly efficient feather-degrading strain,XS6,was isolated,which degraded 97% feather barbs within 24 hours in feather fermentation medium at concentration of 10.0 g/L and exhibited a keratinase activity of 201.2 U/mL.After 36 hours of cultivation,nitrite was detected in the medium.The morphological and molecular biology identification showed that strain XS6 was a Bacillus albus.Transcriptome sequencing showed that a total of 655 DEGs were screened between the two treatment groups,including 148 up-regulated genes and 507 down-regulated genes.GO functional annotation showed that DEGs were annotated to 32 functional groups in 3 categories,among which genes related to metabolic processes,cellular processes,cell,cell parts and catalytic activity were annotated more frequently.KEGG enrichment analysis showed that DEGs were mainly enriched in energy and material metabolism pathways,signal transduction pathways and biosynthetic pathways.【Conclusion】A highly efficient Bacillus albus.strain XS6 capable of degrading feathers was successfully obtained,which participated in synergistic interactions among strains,energy metabolism and biosynthesis of large biomolecules during the degradation process. |
Key words: feather degradation degrading bacteria transcriptomics degradation mechanism |