芳烃诱导Acinetobacter sp. CZ合成靛蓝及染布应用研究

Indole biotransformation and indigo biosynthesis by Acinetobacter sp.CZ stimulated by aromatics and its application in dyeing cloth

靛蓝是一种古老的天然染料,被广泛应用于食品、染色以及医药等行业.利用微生物转化废水中的含氮杂环污染物吲哚合成靛蓝对于污染物的绿色资源化具有重要意义.本研究从焦化废水处理厂二沉池中分离得到靛蓝合成菌CZ,经形态学观察和16S rRNA基因测序鉴定为不动杆菌属(Acinetobacter sp.).采用液相色谱/飞行时间-质谱联用仪(LC/TOF-MS)测定菌株代谢的中间产物,发现靛红、靛红酸及邻氨基苯甲酸是菌株CZ降解吲哚的中间产物,降解过程中有副产物靛蓝生成.同时,条件优化结果表明菌株CZ在受到苯酚及联苯刺激作用下合成靛蓝产量得到显著提升.进一步利用响应曲面法考察菌株CZ合成靛蓝的最佳条件为:吲哚浓度292.88 mg·L^(-1)和苯酚浓度135.89 mg·L^(-1)、吲哚浓度286.83 mg·L^(-1)和联苯518.96 mg·L^(-1).靛蓝产量分别达到34.86 mg·L^(-1)以及29.51 mg·L^(-1),相较于初始条件分别提升12.58倍及10.65倍.将合成的生物靛蓝应用于染布,证明了其具备染色能力,K/S值在3.22~3.45之间.本研究可为探索吲哚污水的资源化合成靛蓝及其应用提供有力的理论支持.

Indigo,an ancient natural dye widely used in the food,dyeing,and medicine industries,holds immense value for green recycling of the nitrogen heterocyclic pollutant,indole,through microorganism-mediated synthesis.In this study,an indigo-producing bacterium CZ was isolated from the secondary sedimentation tank of the coking wastewater treatment plant and identified as Acinetobacter sp.based on morphological observation and 16S rRNA gene sequencing.The intermediate products of indole metabolism were determined by LC/TOF-MS.It was found that isatin,isatin acid,anthranilic acid,as well as the byproduct indigo as the key intermediates.To enhance the production of indigo,condition optimization was conudcted and results showed that the production of indigo by strain CZ was significantly improved under the stimulation of phenol and biphenyl.Response surface method was further used to investigate the optimum conditions for indigo synthesis by strain CZ.The optimal conditions were as follows:indole 292.88 mg·L^(-1),phenol 135.89 mg·L^(-1);indole 286.83 mg·L^(-1),biphenyl 518.96 mg·L^(-1).The indigo yield reached 34.86 mg·L^(-1) and 29.51 mg·L^(-1),which increased 12.58-and 10.65-fold,respectively.The bio-synthetic indigo was successfully applied in dyeing cloth,achieving a K/S value ranging from 3.22 to 3.45.This study provides strong theoretical support for the synthesis of indigo from indole wastewater and its application.