Systematic profiling of indole-3-acetic acid biosynthesis in bacteria using LC-MS/MS.
Indole-3-acetic acid (IAA) is produced from tryptophan through five synthesis pathways. A comprehensive method for the quantification of IAA and biosynthesis-related intermediates in a culture medium was developed. Sample preparation was simple with protein precipitation. The analytes were separated on a superficially porous C18 silica column and detected by electrospray ionization-tandem mass spectrometry in the positive ion multiple reaction monitoring mode. The limit of detection was 0.05 μM, and the lower limits of quantification ranged from 0.05 to 2 μM. The intra-day and inter-day precision and accuracy were less than 13.96%. Ion suppression was observed, and the deuterated internal standards were used to compensate for the matrix effect. The method was applied to analyze changes in tryptophan catabolism in a culture medium of Pseudomonas putida. The proposed method is robust and suitable for the systematic profiling of IAA biosynthesis in culture supernatant.
Department of Microbiology, Tzu Chi University, Hualien, Taiwan.
Guang-Huey
GH
Lin
Institute of Medical Biotechnology, Tzu Chi University, Hualien, Taiwan.
Chung-Yu
CY
Chang
Institute of Medical Biotechnology, Tzu Chi University, Hualien, Taiwan; Department of Laboratory Medicine and Biotechnology, Tzu Chi University, 701 Section 3, Jhongyang Road, Hualien 970, Taiwan. Electronic address: rita0107@mail.tcu.edu.tw.
Huei-Ru
HR
Lin
D002853:Chromatography, Liquid / Q000379:methods*; D007210:Indoleacetic Acids / Q000032:analysis* / Q000378:metabolism*; D057230:Limit of Detection; D016014:Linear Models; D016958:Pseudomonas putida / Q000378:metabolism; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014364:Tryptophan / Q000032:analysis / Q000378:metabolism
D007210:Indoleacetic Acids; C030737:indoleacetic acid; D014364:Tryptophan
IAA biosynthesis; Indole-3-acetic acid; Liquid chromatography–tandem mass spectrometry; Tryptophan