1. 天津商业大学，天津 300134 2. 天津大学，天津 300027 3. Arizona State University, Tempe, Arizona 85287, USA
Rapid Isolation of Phenol Degrading Bacteria by Fourier Transform Infrared (FTIR) Spectroscopy
LI Fei1, SONG Wen-jun1*, WEI Ji-ping2, WANG Su-ying1, LIU Chong-ji3
1. Tianjin University of Commerce, Tianjin 300134, China 2. Tianjin University, Tianjin 300027, China 3. Arizona State University, Tempe, Arizona 85287, USA
摘要： 苯酚是一种重要的化工原料并广泛存在于工业废水中，随着各国对苯酚生物毒性的认识，排放标准日益提高。生物法作为一种高效、 低成本、 不易二次污染的方法常用于含酚的废水处理。但是可降解苯酚的微生物筛选却是一个复杂繁琐的过程。衰减全反射傅里叶红外光谱(attenuated total reflection Fourier transform infrared，ATR-FTIR)技术是一种高效、 快捷、 高指纹特性的物理检测技术，主成分分析联用最小偏二乘法(principal component analysis-partial least squares，PCA-PLS)是一种有效提取特征指纹峰并建立模型的方法，该实验联合ATR-FTIR检测技术和PCA-PLS统计方法建立苯酚浓度与吸光度模型，可以快速检测固体培养基中底物浓度。实验建立模型判定系数可以达到99.5%，预测集的判定系数可以到达99.4%，说明模型具有较高的拟合性和推广性。通过模型可以预测出菌株降解后固体培养基底物浓度，筛选出可降解苯酚功能微生物，传统的液体培养并采用气相色谱检测残留苯酚浓度筛选出的结果与ATR-FTIR方法筛选出结果进行对比发现，得到相同的筛选结果。结果表明ATR-FTIR联合PCA-PLS建立高拟合度模型，可以快速检测固体培养基底物浓度，从而达到快速筛选可降解苯酚菌的目的，这种方法可以应用到其他有特征指纹峰的底物中，ATR-FTIR是一种可以广泛应用到功能微生物筛选的快速检测方法。
关键词：苯酚;衰减全反射傅里叶红外光谱(ATR-FTIR);最小偏二乘(PLS);功能微生物
Abstract：Phenol is an important chemical engineering material and ubiquitous in industry wastewater, its existence has become a thorny issue in many developed and developing country. More and more stringent standards for effluent all over the world with human realizing the toxicity of phenol have been announced. Many advanced biological methods are applied to industrial wastewater treatment with low cost, high efficiency and no secondary pollution, but the screening of function microorganisms is certain cumbersome process. In our study a rapid procedure devised for screening bacteria on solid medium can degrade phenol coupled with attenuated total reflection fourier transform infrared (ATR-FTIR) which is a detection method has the characteristics of efficient, fast, high fingerprint were used. Principal component analysis (PCA) is a method in common use to extract fingerprint peaks effectively, it couples with partial least squares (PLS) statistical method could establish a credible model. The model we created using PCA-PLS can reach 99.5% of coefficient determination and validation data get 99.4%, which shows the promising fitness and forecasting of the model. The high fitting model is used for predicting the concentration of phenol at solid medium where the bacteria were grown. The highly consistent result of two screening methods, solid cultural with ATR-FTIR detected and traditional liquid cultural detected by GC methods, suggests the former can rapid isolate the bacteria which can degrade substrates as well as traditional cumbersome liquid cultural method. Many hazardous substrates widely existed in industry wastewater, most of them has specialize fingerprint peaks detected by ATR-FTIR, thereby this detected method could be used as a rapid detection for isolation of functional microorganisms those can degrade many other toxic substrates.
Key words：Industrial sewage;Phenol biodegradation;Screening;ATR-FTIR;PLS model
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