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一株耐热耐冷漆酶产生菌的特性研究及其在酶促氧化降解土霉素和金霉素中的应用。

Characterization of a robust cold-adapted and thermostable laccase from Pycnoporus sp. SYBC-L10 with a strong ability for the degradation of tetracycline and oxytetracycline by laccase-mediated oxidation.

机构信息

The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu, 214122, PR China.

The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu, 214122, PR China.

出版信息

J Hazard Mater. 2020 Jan 15;382:121084. doi: 10.1016/j.jhazmat.2019.121084. Epub 2019 Aug 22.

Abstract

A native laccase (Lac-Q) with robust cold-adapted and thermostable characteristics from the white-rot fungus Pycnoporus sp. SYBC-L10 was purified, characterized, and used in antibiotic treatments. Degradation experiments revealed that Lac-Q at 10.0 U mL coupled with 1.0 mmol L ABTS could degrade 100% of the tetracycline or oxytetracycline (50 mg L) within 5 min with a static incubation at 0 °C (pH 6.0). The presence of the Mn ion inhibited the removal rate of tetracycline and oxytetracycline by the Lac-Q-ABTS system, and the presence of Al, Cu, and Fe accelerated the removal rate of tetracycline and oxytetracycline by the Lac-Q-ABTS system. Furthermore, the growth inhibition of Bacillus altitudinis SYBC hb4 and E. coli by tetracycline antibiotics revealed that the antimicrobial activity was significantly reduced after treatment with the Lac-Q-ABTS system. Finally, seven transformation products of oxytetracycline (namely TP 445, TP 431, TP 413, TP 399, TP 381, TP 367, and TP 351) were identified during the Lac-Q-mediated oxidation process by using UPLC-MS/MS. A possible degradation pathway including deamination, demethylation, and dehydration was proposed. These results suggest that the Lac-Q-ABTS system shows a great potential for the treatment of antibiotic wastewater containing different metal ions at various temperatures.

摘要

从白腐真菌 Pycnoporus sp. SYBC-L10 中纯化、表征了一种具有强大耐冷和耐热特性的天然漆酶 (Lac-Q),并将其用于抗生素处理。降解实验表明,在 0°C(pH 6.0)下静态孵育时,10.0 U mL 的 Lac-Q 与 1.0 mmol L ABTS 偶联可以在 5 分钟内降解 100%的四环素或土霉素(50 mg L)。Mn 离子的存在抑制了 Lac-Q-ABTS 系统对四环素和土霉素的去除率,而 Al、Cu 和 Fe 的存在则加速了 Lac-Q-ABTS 系统对四环素和土霉素的去除率。此外,四环素抗生素对高山芽孢杆菌 SYBC hb4 和大肠杆菌的生长抑制作用表明,在用 Lac-Q-ABTS 系统处理后,其抗菌活性显著降低。最后,通过使用 UPLC-MS/MS,在 Lac-Q 介导的氧化过程中鉴定出土霉素的 7 种转化产物(即 TP 445、TP 431、TP 413、TP 399、TP 381、TP 367 和 TP 351)。提出了包括脱氨、去甲基化和脱水在内的可能降解途径。这些结果表明,Lac-Q-ABTS 系统在处理不同温度下含有不同金属离子的抗生素废水方面具有很大的潜力。

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