A commercial laccase from Trametes versicolor was conjugated with biopolymer chitosan using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) as the cross-linking agent. Laccase-chitosan conjugation strategies were tested using different molar ratios of glucosamine monomer/protein with different molar excess ratios of EDC relative to laccase. Immobilization techniques were developed to improve the stability against thermal and chemical denaturation, storage and reusability of this biocatalyst. The conjugation resulted in a solid biocatalyst with an apparent laccase activity of ±626 U/g, 12 and 60 folds higher in the conjugation efficiency of biocatalyst relative to the immobilized and free laccase activity respectively when compared with zero EDC/laccase ratio used in conjugation solution. The conjugated laccases formed successfully eliminated the emerging pollutant triclosan (TCS) from aqueous solutions, having a higher potential to transform TCS than free laccase. UPLC-QTOF results indicate the formation of TCS oligomers. Furthermore, they are the first evidence of direct dechlorination of TCS mediated by the oxidative action of laccases.

译文

使用1-乙基-3-(3-二甲基氨基丙基) 碳二亚胺盐酸盐 (EDC) 作为交联剂,将versicolor的市售漆酶与生物聚合物壳聚糖偶联。使用不同的葡萄糖胺单体/蛋白质摩尔比以及相对于漆酶的EDC摩尔过量比,测试了漆酶-壳聚糖结合策略。开发了固定化技术,以提高该生物催化剂对热和化学变性,存储和可重复使用性的稳定性。与用于缀合溶液的零EDC/漆酶比相比,该缀合产生了表观漆酶活性为 ± 626 U/g的固体生物催化剂,与固定漆酶活性和游离漆酶活性相比,生物催化剂的缀合效率分别高12和60倍。形成的共轭漆酶成功地消除了水溶液中出现的污染物三氯生 (TCS),比游离漆酶具有更高的转化TCS的潜力。Uplc-qtof结果表明形成了TCS低聚物。此外,它们是漆酶氧化作用介导的TCS直接脱氯的第一个证据。

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