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根据全基因组测序和转录组分析,解析海洋红树林芽孢杆菌 NM1-A2 的生物脱硫途径。

Deciphering the biodesulfurization pathway employing marine mangrove Bacillus aryabhattai strain NM1-A2 according to whole genome sequencing and transcriptome analyses.

机构信息

National Engineering Research Center for Non-Food Biorefinery, State Key Laboratory of Non-Food Biomass and Enzyme Technology, Guangxi Key Laboratory of Bio-refinery, Guangxi Research Center for Biological Science and Technology, Guangxi Academy of Sciences, Nanning 530007, China; State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi Research Center for Microbial and Enzyme Engineering Technology, College of Life Science and Technology, Guangxi University, Nanning 530004, China.

State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi Research Center for Microbial and Enzyme Engineering Technology, College of Life Science and Technology, Guangxi University, Nanning 530004, China.

出版信息

Genomics. 2023 May;115(3):110635. doi: 10.1016/j.ygeno.2023.110635. Epub 2023 May 6.

Abstract

In the biogeochemical cycle, sulfur oxidation plays a vital role and is typically referred to as the elemental sulfur or reductive sulfide oxidation process. This study aimed to characterize a subtropical mangrove-isolated bacterial strain using biochemical, whole-genome, and transcriptome sequencing analyses to enhance our understanding of sulfur metabolism and biodegradation from a molecular genetic perspective. Strain NM1-A2 was characterized as Gram-positive and found to have a close molecular phylogenetic relationship with Bacillus aryabhattai. NM1-A2 efficiently converted dibenzothiophene (DBT) into 2-hydroxybiphenyl (2-HBP) via a 4S pathway with 95% efficiency, using enzymes encoded by the dsz operon (dszA, dszB, and dszC), which determine monooxygenases (DszA & DszC) and desulfinase (DszB). The whole-genome sequence of NM1-A2 had a length of approximately 5,257,678 bp and included 16 sulfur metabolism-related genes, featuring the ABC transport system, small subunit (ssu) and cysteine (cys) gene families, and adenosine 5'-phosphosulfate (APS) and 3'-phosphoadenosine-5'-phosphosulfate (PAPS) biosynthesis-related genes. Transcriptomic analysis of NM1-A2 using three sulfur groups-magnesium sulfate (MS), sulfur powder (SP), and sodium thiosulfate (ST) resulted in a significant number of differentially expressed genes (1200, 2304, and 2001, respectively). This analysis revealed that intracellular cysteine concentration directly regulated the expression of cys and ssu genes. Sulfate did not directly affect cys gene expression but repressed ssu gene expression. The cys gene expression levels decreased during the conversion of sulfate to sulfide and cysteine. The transcriptomic data was validated by analyzing the expression patterns of NM1-A2 using real-time quantitative PCR validation analysis. The expression levels of cysl, mccB, and nrnA were significantly upregulated, while cysH, metB, and sat were downregulated in the SP, ST, and MS groups, respectively. This research contributes to our understanding of marine mangrove microorganisms' bacterial efficiency through characterization, whole-genome, and transcriptome sequencing-based molecular degradation of organic compounds in the mangrove ecosystem, which may enhance nutrient availability.

摘要

在生物地球化学循环中,硫氧化起着至关重要的作用,通常被称为元素硫或还原性硫化物氧化过程。本研究旨在通过生化、全基因组和转录组测序分析来表征一株亚热带红树林分离的细菌菌株,以从分子遗传学的角度增强我们对硫代谢和生物降解的理解。菌株 NM1-A2 被鉴定为革兰氏阳性,与 Bacillus aryabhattai 具有密切的分子系统发育关系。NM1-A2 通过 4S 途径以 95%的效率将二苯并噻吩(DBT)有效地转化为 2-羟基联苯(2-HBP),该途径使用 dsz 操纵子(dszA、dszB 和 dszC)编码的酶,这些酶决定单加氧酶(DszA 和 DszC)和脱硫酶(DszB)。NM1-A2 的全基因组序列约为 5,257,678 bp,包含 16 个与硫代谢相关的基因,具有 ABC 转运系统、小亚基(ssu)和半胱氨酸(cys)基因家族,以及腺苷 5'-磷酸硫酸(APS)和 3'-磷酸腺苷-5'-磷酸硫酸(PAPS)生物合成相关基因。使用三种硫源-硫酸镁(MS)、硫磺粉(SP)和硫代硫酸钠(ST)对 NM1-A2 进行转录组分析,结果产生了大量差异表达基因(分别为 1200、2304 和 2001 个)。该分析表明,细胞内半胱氨酸浓度直接调节 cys 和 ssu 基因的表达。硫酸盐不直接影响 cys 基因的表达,但抑制 ssu 基因的表达。在硫酸盐转化为硫化物和半胱氨酸的过程中,cys 基因的表达水平下降。通过使用实时定量 PCR 验证分析对 NM1-A2 的表达模式进行分析,验证了转录组数据。在 SP、ST 和 MS 组中,cysl、mccB 和 nrnA 的表达水平显著上调,而 cysH、metB 和 sat 的表达水平下调。这项研究通过对红树林生态系统中有机化合物的分子降解进行特征描述、全基因组和转录组测序,有助于我们了解海洋红树林微生物的细菌效率,这可能会增强营养物质的可利用性。

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