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一项关于塑料添加剂2,6-二叔丁基苯酚生物降解条件及降解菌三爱蛋白质组变化的综合研究。

A comprehensive study of conditions of the biodegradation of a plastic additive 2,6-di--butylphenol and proteomic changes in the degrader san ai.

作者信息

Medić Ana, Stojanović Ksenija, Izrael-Živković Lidija, Beškoski Vladimir, Lončarević Branka, Kazazić Saša, Karadžić Ivanka

机构信息

Department of Chemistry, Faculty of Medicine, University of Belgrade Višegradska 26 11000 Belgrade Serbia

Faculty of Chemistry, University of Belgrade Studentski trg 12-16 11000 Belgrade Serbia.

出版信息

RSC Adv. 2019 Jul 30;9(41):23696-23710. doi: 10.1039/c9ra04298a. eCollection 2019 Jul 29.

Abstract

The san ai strain was investigated for its capability to degrade the 2,6-di--butylphenol (2,6-DTBP) plastic additive, a hazardous and toxic substance for aquatic life. This investigation was performed under different parameter values: 2,6-DTBP concentration, inoculum size, pH, and temperature. The GC-MS study showed that efficiently degraded 2,6-DTBP in the pH range of 5-8 at higher temperatures. Under exposure to 2,6-DTBP concentrations of 2, 10, and 100 mg L, the strain degraded by 100, 100, and 85%, respectively, for 7 days. Crude enzyme preparation from the biomass of san ai showed higher efficiency in 2,6-DTBP removal than that shown by whole microbial cells. Gene encoding for the enzymes involved in the degradation of aromatic compounds in san ai was identified. To complement the genomic data, a comparative proteomic study of san ai grown on 2,6-DTBP or sunflower oil was conducted by means of nanoLC-MS/MS. The presence of aromatic substances resulted in the upregulation of aromatic ring cleavage enzymes, whose activity was confirmed by enzymatic tests; therefore, it could be concluded that 2,6-DTBP might be degraded by -ring cleavage. A comparative proteomics study of san ai indicated that the core molecular responses to aromatic substances can be summarized as the upregulation of proteins responsible for amino acid metabolism with emphasized glutamate metabolism and energy production with upregulated enzymes of glyoxylate bypass. san ai has a high capacity to efficiently degrade aromatic compounds, and therefore its whole cells or enzymes could be used in the treatment of contaminated areas.

摘要

研究了三爱菌株降解2,6 - 二叔丁基苯酚(2,6 - DTBP)塑料添加剂的能力,该物质对水生生物具有有害毒性。在不同参数值下进行了此项研究:2,6 - DTBP浓度、接种量、pH值和温度。气相色谱 - 质谱联用(GC - MS)研究表明,在较高温度下,该菌株能在pH值为5 - 8的范围内有效降解2,6 - DTBP。在暴露于浓度为2、10和100 mg/L的2,6 - DTBP环境中7天后,该菌株对其降解率分别为100%、100%和85%。从三爱菌株的生物质中制备的粗酶制剂在去除2,6 - DTBP方面比完整的微生物细胞表现出更高的效率。鉴定出了三爱菌株中参与芳香化合物降解的酶的编码基因。为补充基因组数据,通过纳升液相色谱 - 串联质谱(nanoLC - MS/MS)对在2,6 - DTBP或向日葵油上生长的三爱菌株进行了比较蛋白质组学研究。芳香物质的存在导致芳香环裂解酶上调,其活性通过酶促试验得到证实;因此,可以得出结论,2,6 - DTBP可能通过β - 环裂解进行降解。对三爱菌株的比较蛋白质组学研究表明,其对芳香物质的核心分子反应可概括为负责氨基酸代谢(尤其是谷氨酸代谢)的蛋白质上调,以及乙醛酸循环旁路酶上调导致的能量产生增加。三爱菌株具有高效降解芳香化合物的高能力,因此其完整细胞或酶可用于污染区域的处理。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a2de/9069449/fc80316cbb9f/c9ra04298a-f1.jpg

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