齿毛菌漆酶对活性亮蓝的高效脱色
江 聪1,尹大强2,许鑫琦1,林 娟1,叶秀云1,杨 捷1*

(1.福州大学福建省海洋酶工程重点实验室,福建 福州 350116; 2.同济大学长江水环境教育部重点实验室,上海 200092)

漆酶; 活性亮蓝; 脱色; 响应面法; 红外光谱; 薄层色谱

Efficient Decolorization of Remazol Brilliant Blue R by Cerrena unicolor Laccase
JIANG Cong1,YIN Daqiang2,XU Xinqi1,LIN Juan1,YE Xiuyun1,YANG Jie1*

(1.Fujian Key Laboratory of Marine Enzyme Engineering,Fuzhou University,Fuzhou 350116,China; 2.Key Laboratory of Yangtze River Water Environment,Ministry of Education,Tongji University,Shanghai 200092,China)

laccase; Remazol Brilliant Blue R; decolorization; response surface method; infrared spectrometry; thin layer chromatography

DOI: 10.6043/j.issn.0438-0479.201707010

备注

漆酶是一种含铜的多酚氧化酶,在环境污染治理等领域具有广阔的应用前景.采用齿毛菌(Cerrena unicolor)漆酶对蒽醌染料活性亮蓝(Remazol Brilliant Blue R,RBBR)进行催化脱色.在单因素试验的基础上,应用中心组合设计(central composite design,CCD)建立4因素3水平的响应面法分析模型,得到齿毛菌漆酶对RBBR脱色的最佳条件:漆酶浓度2.24 U/mL,染料质量浓度123.83 mg/L,pH 4.52,脱色时间38.30 min,预测理论脱色率为99.82%.RBBR在最佳脱色条件下的实际脱色率为(98.42±1.39)%.影响脱色效果的因素主次顺序为:pH>酶浓度>脱色时间>染料质量浓度.红外光谱和薄层色谱分析发现齿毛菌漆酶处理后RBBR的结构发生改变,其中的C—N键发生断裂,证明该漆酶确实催化了RBBR的降解.齿毛菌漆酶对RBBR脱色反应的米氏常数Km值为(134.82 ±3.05)mg/L,不需要氧化还原介体就可以对RBBR进行高效脱色,在染料脱色和生物修复方面具有实际应用价值.

Laccases are members of a family of copper-containing polyphenol oxidases and have wide applications in bioremediation.In this study,catalytic decolorization of Remazol Brilliant Blue R(RBBR)by Cerrena unicolor laccase was investigated.Based on the single factor experiments,the central composite design(CCD)was used to establish the four-factor three-level response surface method analysis model,and the optimum conditions of laccase-mediated RBBR decolorization were as follows:2.24 U/mL laccase,123.83 mg/L RBBR,pH 4.52,and 38.30 min.The predicted and observed decolorization rates were 99.82% and(98.42±1.39)%,respectively,under the optimal conditions.Influences of the factors on the decolorization efficiency were pH> laccase concentration> decolorization time> dye concentration.The infrared spectrometry and thin layer chromatography analyses showed that the structure of RBBR changed due to the laccase treatment.Specifically,the C—N bond in RBBR was broken upon laccase treatment,corroborating that the laccase catalyzed RBBR degradation.The Km value of the C. unicolor laccase towards RBBR was(134.82±3.05)mg/L,as indicated by the kinetic study,and efficiently decolorized RBBR in the absence of a redox mediator,showing promise in dye decolorization and bioremediation.