苯酚羟化酶生物转化吲哚及甲基吲哚研究  

Research on biotransformation of indole and methylindoles by phenol hydroxylase

作　　者：张旭旺[1] 张强[1] 曲媛媛[1] 时胜男[2] 许炳雯[1] 马桥[1] 沈娥[1] 周集体[1] 

机构地区：[1]大连理工大学工业生态与环境工程教育部重点实验室,辽宁大连116024 [2]哈尔滨工业大学城市水资源与水环境国家重点实验室,黑龙江哈尔滨150090

出　　处：《大连理工大学学报》2013年第3期346-353,共8页Journal of Dalian University of Technology

基　　金：国家自然科学基金资助项目(51078054;21176040;20923006)

摘　　要：利用苯酚羟化酶基因工程菌PH_IND对吲哚及甲基吲哚进行生物转化.结果表明,除了3-甲基吲哚外,PH_IND能将吲哚、2-甲基吲哚、4-甲基吲哚和7-甲基吲哚转化成靛蓝类色素.采用薄层色谱和液相-质谱等手段对转化产物进行鉴定,结果表明,PH_IND转化吲哚和4-甲基吲哚得到的蓝色产物分别为靛蓝和4,4′-二甲基靛蓝,并推测了PH_IND转化吲哚及4-甲基吲哚的反应途径.采用表面响应法对PH_IND转化吲哚、2-甲基吲哚及4-甲基吲哚的条件进行了优化,在最优条件下,菌株PH_IND转化吲哚合成靛蓝的最大产量为93.74mg/L,其吲哚的转化效率为52.63%,且菌株PH_IND对2-甲基吲哚和4-甲基吲哚的最大转化率分别为67.66%和95.61%.The gene engineering strain PH_IND expressing phenol hydroxylase is used to catalyze the biotransformation of indole and methylindoles. The experimental results indicate that indole, 2-methylindole, 4-methylindole and 7-methylindole can be transformed into various indigoid pigments except for 3-methylindole. The predominant color products are identified by thin-layer chromatography and liquid chromatography-mass spectrometry analyses, and the major products （blue pigment） derived from indole and 4-methylindole are authenticated as indigo and 4, 4＇-dimethylindigo,respectively. Meanwhile, the biotransformation pathways of indole and 4-methylindole by PH_IND areproposed. Response surface methodology is applied to optimize the biotransformation conditions of indole, 2-methylindole and 4-methylindole by PH_IND. Under the optimal conditions, the indigo yield and transformation efficiency of indole are 93. 74 mg/L and 52.63%, respectively, while the maximum transformation rates of 2-methylindole and 4-methylindole are 67. 66% and 95. 61%,respectively.

关 键 词：吲哚 甲基吲哚 苯酚羟化酶 表面响应法 

分 类 号：Q814[生物学—生物工程] X172[环境科学与工程—环境科学]