范丽莉 王炎 赵敏 王珏玉 赵美 邢丹

(哈尔滨工业大学，哈尔滨，150001) (东北林业大学)



CotA漆酶催化靛蓝胭脂红脱色条件优化1)

范丽莉 王炎 赵敏 王珏玉 赵美 邢丹

(哈尔滨工业大学，哈尔滨，150001) (东北林业大学)

采用Plackett-Burman试验设计法及响应面分析优化了重组细菌漆酶CotA催化靛蓝胭脂红脱色的条件。经过Plackett-Burman试验筛选出影响脱色条件的主要因素为pH、温度和染料质量浓度。利用Box-Behnken中心组合试验设计及响应面分析，通过三元二次数学模型进行回归拟合，预测了靛蓝胭脂红染料最佳脱色条件为：pH=8.6，温度60 ℃，染料质量浓度41.5 mg·L-1，染料在最优方案条件下45 min内的脱色率实际检测值为91.40%，与预测值的误差仅为0.86%。以上结果显示，CotA漆酶在靛蓝胭脂红脱色领域具有较好的应用前景。对脱色产物进行液相质谱分析，检测到m/z=216.07 [M-Na]-的脱色产物，推测该降解产物为4-氨基-3-羧基苯磺酸钠，证明CotA漆酶催化靛蓝胭脂红降解。

CotA；靛蓝胭脂红；染料脱色；响应面优化

The decolorization of Indigo carmine by the CotA laccase was detected. The decolorization conditions were optimized by the Plackett-Burman test design method and response surface method. The Plackett Burman experiment design was conducted to select the relatively important factors in indigo carmine decolorization. The temperature, pH and concentration of dye were the three significant factors influencing the dye decolorization. Using Box-Behnken center combination experiment design and response surface analysis, the optimal operating condition was with pH of 8.6, 60 ℃ and 41.5 mg·L-1indigo carmine, and the dye decolorization rate was 91.40% within 45 min which was only with 0.86% difference from the forecast value. The results indicated that the promising prospects of the recombinant CotA laccase in indigo carmine decolorization. The molecular ions (m/z=216.07[M-Na]-) were obtained and identified by LC/MS. Therefore, the indigo carmine was degraded to 4-Amino-3-carboxybenzenesulfonic acid sodium salt and the indigo carmine could be degraded by the CotA laccase.

靛蓝胭脂红(IC)是一种应用非常广泛的靛蓝类染料，可用于牛仔裤和其他蓝色织物的染色，靛蓝胭脂红还可以作为分析化学中指示剂以及用于显微镜观察的染色剂[1]。此外，靛蓝胭脂红在食品、药品和化妆品工业中都有所应用[2]。然而这类靛蓝染料可以引发肿瘤致癌，并可导致生殖、发育和神经性疾病，威胁人体健康[3]，因此脱色降解水体中的靛蓝胭脂红对于减小该染料造成的污染至关重要。目前已有一些利用物理吸附的方法去除废水中靛蓝胭脂红的报道，使用的吸附材料包括壳聚糖[4]和活性炭[5]等。……