Huang Wei E, Hopper David, Goodacre Royston, Beckmann Manfred, Singer Andrew, Draper John
Molecular Microbial Ecology Laboratory, CEH-Oxford, Mansfield Road, Oxford, OX1 3SR, UK.
J Microbiol Methods. 2006 Nov;67(2):273-80. doi: 10.1016/j.mimet.2006.04.009. Epub 2006 Jun 5.
Fourier transform-infrared (FT-IR) spectroscopy has become an important tool for rapid analysis of complex biological samples. The infrared absorbance spectrum could be regarded as a "fingerprint" which is characteristic of biochemical substances. In this study, Pseudomonas putida NCIMB 9869 was grown with either 3,5-xylenol or m-cresol as the sole carbon source, each inducing different metabolic pathways for m-cresol biotransformation. FT-IR spectroscopy was capable of differentiating both induced cultures of P. putida NCIMB 9869 as well as the resulting biotransformation product mixtures. FT-IR spectral analysis indicated that carboxylic acids were key chemicals responsible for distinguishing the products of the two catabolic pathways. Gas chromatography-mass spectrometry (GC-MS) was performed to validate the FT-IR analysis, indicating that two carboxylic acids, 3-hydroxybenzoic acid and 2,5-dihydroxybenzoic acid, were present as m-cresol biotransformation products from 3,5-xylenol-grown cells, but were absent in m-cresol-grown cells. The ability to use FT-IR to rapidly distinguish between biotransformation product mixtures as well as differentially induced bacterial strains suggests this approach might be a valuable tool for screening large biotransformation assays for novel products and metabolic mutants.
傅里叶变换红外（FT-IR）光谱法已成为快速分析复杂生物样品的重要工具。红外吸收光谱可被视为生化物质特有的“指纹”。在本研究中，恶臭假单胞菌NCIMB 9869以3,5-二甲苯酚或间甲酚作为唯一碳源进行培养，每种碳源诱导间甲酚生物转化的代谢途径不同。FT-IR光谱法能够区分恶臭假单胞菌NCIMB 9869的两种诱导培养物以及由此产生的生物转化产物混合物。FT-IR光谱分析表明，羧酸是区分两种分解代谢途径产物的关键化学物质。进行气相色谱-质谱联用（GC-MS）以验证FT-IR分析，结果表明，两种羧酸，即3-羟基苯甲酸和2,5-二羟基苯甲酸，作为以3,5-二甲苯酚培养的细胞间甲酚生物转化产物存在，但在以间甲酚培养的细胞中不存在。利用FT-IR快速区分生物转化产物混合物以及差异诱导的细菌菌株的能力表明，这种方法可能是筛选新型产物和代谢突变体的大型生物转化试验的有价值工具。
