伽马辐照后铜绿假单胞菌氮素基因表达的检测和分子深入研究

IF 2.3 3区 生物学 Q3 MICROBIOLOGY Archives of Microbiology Pub Date : 2024-11-22 DOI:10.1007/s00203-024-04195-x
Ebthag M. Awad, Nagwa A. Abdallah, Mona M. K. Shehata, Hala A. Farrag
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引用次数: 0

摘要

铜绿假单胞菌(Pseudomonas aeruginosa)产生的一种次生代谢产物--氮杂环苷(Azurin),因其宝贵的治疗和生物应用价值而备受关注。这项研究旨在利用不同剂量的伽马辐照(5-400 Gy),研究铜绿假单胞菌最大限度地产生天青苷的过程。76株铜绿假单胞菌分离物来自135个环境样本和35个临床细菌分离物,从高到低的顺序如下:35 个分离株(46%)来自临床样本,26 个分离株(34%)来自水样本,15 个分离株(20%)来自土壤样本。抗菌药敏感性测试采用了盘扩散技术,结果显示,根据临床和实验室标准研究所(CLSI)确定的标准,所有收集到的分离物中耐多药(MDR)率为 54(71%)。基因组实验结果表明,只有 37 个 MDR 分离物的天青霉素基因检测呈阳性,PCR 产物为 446 bp。傅立叶变换红外光谱分析进一步证实了这些发现,该分析显示在 1636.96 cm- 1 附近出现了峰值,表明这些分离物中的氮杂环苷具有突出的 α-螺旋二级结构。与致病性和抗生素耐药性有关的是,来自临床的分离物表现出较高的氮素基因表达水平。此外,本研究还证实,50 和 100 Gy 的伽马射线照射能显著提高三种受测临床分离物的天青霉素表达水平(P ≤ 0.05),与未受照射的样本相比,最大表达倍数为 63.55。总之,低剂量伽马辐照能有效提高次生代谢产物氮杂嘌呤的表达水平,为生物和医学应用中的后续纯化过程提供了极大的益处。
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Detection and molecular insights into the azurin gene expression post– gamma irradiation in P. aeruginosa

Azurin, a secondary metabolite from Pseudomonas aeruginosa, has attracted much attention owing to its valuable therapeutic and biological applications. This work aimed to study and chartly maximize the azurin production process using different doses of gamma irradiation (5–400 Gy) in P. aeruginosa isolates. Seventy-six P. aeruginosa isolates were sourced from 135 environmental samples and 35 clinical bacterial isolates with the following descending order: 35 isolates (46%) from clinical samples, 26 isolates (34%) from water samples, and 15 isolates (20%) from soil samples. The disc diffusion technique was used for antimicrobial susceptibility testing, revealing that the multidrug-resistant (MDR) rate among all collected isolates according to the criteria determined by Clinical and Laboratory Standards Institute (CLSI) was 54 (71%). The genomic experimental results revealed that only 37 MDR isolates tested positive for the azurin gene, as detected by the PCR product at 446 bp. These findings were further supported by FTIR analysis, which revealed peaks around 1636.96 cm− 1, indicating a prominent α-helix secondary structure of azurin in these isolates. Related to their pathogenicity and antibiotic resistance, isolates from clinical origin exhibited the higher azurin gene expression level. Besides, this study confirmed the potency of gamma radiation exposure at 50 and 100 Gy significantly increased the azurin expression levels in three tested clinical isolates (P ≤ 0.05), with a maximum fold expression level of 63.55 compared to the non-irradiated samples. In conclusion, low doses of gamma irradiation effectively enhanced expression level of a secondary metabolite azurin, providing a considerable benefit for subsequent purification processes in both biological and medical applications.

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来源期刊
Archives of Microbiology
Archives of Microbiology 生物-微生物学
CiteScore
4.90
自引率
3.60%
发文量
601
审稿时长
3 months
期刊介绍: Research papers must make a significant and original contribution to microbiology and be of interest to a broad readership. The results of any experimental approach that meets these objectives are welcome, particularly biochemical, molecular genetic, physiological, and/or physical investigations into microbial cells and their interactions with their environments, including their eukaryotic hosts. Mini-reviews in areas of special topical interest and papers on medical microbiology, ecology and systematics, including description of novel taxa, are also published. Theoretical papers and those that report on the analysis or ''mining'' of data are acceptable in principle if new information, interpretations, or hypotheses emerge.
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