Fourier Transform Infrared Spectroscopy for Screening and Quantifying Production of PHAs by Pseudomonas Grown on Sodium Octanoate

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Abstract

Poly(hydroxyalkanoates) PHAs are synthesized by many bacteria as inclusion bodies and their biodegradability and structural diversity have been studied with a view to their potential application as biodegradable materials. A method based on FT-IR was developed to carry out rapid qualitative and quantitative analysis of PHAs in Pseudomonas, when they were grown on sodium octanoate. Using absorbance of the ester band of PHAs, a rapid method was reported to distinguish PHB and PHO and to determine polymer content in intact bacteria. Relative areas in which the C=O area was normalized to the area of the peak representing the amid group (1656 cm(-1)) characteristic of bacteria were calibrated to the polymer content which was determined after solvent extraction. Polymer contents vary from 0% to 53% and depend on the nature of the bacteria. Among 27 strains of Pseudomonas belonging to the rRNA homology group I, a very low amount of bacteria were able to produce PHB. The majority of strains were able to produce a copolymer, PHO, in which the major constituent unit is 3-hydroxyoctanoate. The FT-IR results were further confirmed by gas chromatography analysis after methanolysis of polymer, but FT-IR method requires less preparation of sample than gas chromatography and it is very useful for screening a large variety of Pseudomonas.

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