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使用化学信息学、体内和体外实验对细胞毒性和生物活性进行研究,利用新的表型和基因型特征明确的2株链霉菌菌株对乙酰氨基酚痕量进行生物修复。

Acetaminophen-traces bioremediation with novel phenotypically and genotypically characterized 2 Streptomyces strains using chemo-informatics, in vivo, and in vitro experiments for cytotoxicity and biological activity.

作者信息

Embarez Donia H, Razek Ahmed S Abdel, Basalious Emad B, Mahmoud Magdi, Hamdy Nadia M

机构信息

Biochemistry Department, Faculty of Science, Ain Shams University, Cairo, 11566, Abassia, Egypt.

Microbial Chemistry Department, Genetic Engineering and Biotechnology Research Division, National Research Centre, Giza, 12622, Dokki, Egypt.

出版信息

J Genet Eng Biotechnol. 2023 Dec 19;21(1):171. doi: 10.1186/s43141-023-00602-w.

Abstract

We isolated two novel bacterial strains, active against the environmental pollutant acetaminophen/Paracetamol®. Streptomyces chrestomyceticus (symbol RS2) and Flavofuscus (symbol M33) collected from El-Natrun Valley, Egypt-water, sediment, and sand samples, taxonomically characterized using a transmission electron microscope (TEM). Genotypic identification, based on 16S rRNA gene sequence analysis followed by BLAST alignment, were deposited on the NCBI as 2 novel strains https://www.ncbi.nlm.nih.gov/nuccore/OM665324  and https://www.ncbi.nlm.nih.gov/nuccore/OM665325 . The phylogenetic tree was constructed. Acetaminophen secondary or intermediate product's chemical structure was identified by GC/LC MS. Some selected acetaminophen secondary-product extracts and derived compounds were examined against a panel of test micro-organisms and fortunately showed a good anti-microbial effect. In silico chemo-informatics Swiss ADMET evaluation was used in the selected bio-degradation extracts for absorption (gastric), distribution (to CNS), metabolism (hepatic), excretion (renal), and finally not toxic, being non-mutagenic/teratogenic or genotoxic, virtually. Moreover, in vitro cytotoxic activity of these selected bio-degradation secondary products was examined against HepG2 and MCF7 cancer cell lines, where M33 and RS2 extract effects on acetaminophen/paracetamol bio-degradation products were safe, with higher IC50 on HepG2 and MCF7 than the acetaminophen/paracetamol IC50 of 108.5 μg/ml. Moreover, an in vivo oral acute single-dose toxicity experiment was conducted, to confirm these in vitro and in silico lower toxicity (better safety) than acetaminophen/paracetamol.

摘要

我们分离出了两种对环境污染物对乙酰氨基酚/必理通®具有活性的新型细菌菌株。从埃及纳特龙湖谷的水、沉积物和沙子样本中采集的克雷氏链霉菌(代号RS2)和黄褐黄杆菌(代号M33),使用透射电子显微镜(TEM)进行了分类学表征。基于16S rRNA基因序列分析并经BLAST比对的基因型鉴定结果已作为两种新型菌株存入NCBI,网址分别为https://www.ncbi.nlm.nih.gov/nuccore/OM665324https://www.ncbi.nlm.nih.gov/nuccore/OM665325 。构建了系统发育树。通过GC/LC MS鉴定了对乙酰氨基酚二级或中间产物的化学结构。对一些选定的对乙酰氨基酚二级产物提取物及其衍生化合物进行了针对一组测试微生物的检测,幸运的是显示出了良好的抗菌效果。在选定的生物降解提取物中使用了计算机化学信息学瑞士ADMET评估,以评估其吸收(胃部)、分布(至中枢神经系统)、代谢(肝脏)、排泄(肾脏)情况,最终结果表明其实际上无毒,无致突变/致畸或基因毒性。此外,还检测了这些选定的生物降解二级产物对HepG2和MCF7癌细胞系的体外细胞毒性活性,其中M33和RS2提取物对乙酰氨基酚/对乙酰氨基酚生物降解产物的影响是安全的,其对HepG2和MCF7的IC50高于对乙酰氨基酚/对乙酰氨基酚的IC50(108.5 μg/ml)。此外,还进行了体内口服急性单剂量毒性实验,以确认这些产物在体外和计算机模拟实验中比乙酰氨基酚/对乙酰氨基酚具有更低的毒性(更好的安全性)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3183/10730784/43c4761603a5/43141_2023_602_Fig1_HTML.jpg

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