A rapid method for detecting bacterial polyhydroxyalkanoates in intact cells by Fourier transform infrared spectroscopy |
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| Authors: | K. Hong S. Sun W. Tian G. Q. Chen W. Huang |
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| Affiliation: | (1) Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, China e-mail: microbes@public3.bta.net.cn Tel.: +8610-62783844 Fax: +8610-62788784, CN;(2) Analysis Centre, Tsinghua University, Beijing 100084, China, CN;(3) Microbiology Department, Nanjing Agriculture University, Nanjing 210095, China, CN |
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| Abstract: | Polyhydroxyalkanoates (PHA) 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. In this paper, Fourier-transform infrared spectroscopy (FT-IR) was used to carry out rapid qualitative analysis of PHA in intact bacterial cells. The FT-IR spectra of pure PHA containing short-chain-length monomers, such as hydroxybutyrate (HB), medium-chain-length hydroxyalkanoate (mclHA) monomers including hydroxyoctanoate (HO) and hydroxydecanoate (HD), or both HB and mclHA monomers, showed their strong characteristic band at 1728 cm−1, 1740 cm−1 or 1732 cm−1 respectively. Other accompanying bands near 1280 cm−1 and 1165 cm−1 helped identify the types of PHA. The intensity of the methylene band near 2925 cm−1 provided additional information for PHA characterization. In comparison, bacterial cells accumulating the above PHA also showed strong marker bands at 1732 cm−1, 1744 cm−1 or 1739 cm−1, corresponding to intracellular PHB, mclPHA and P(HB + mclHA) respectively. The accompanying bands visible in pure PHA were also observable in the intact cells. The FT-IR results were further confirmed by gas chromatography analysis. Received: 14 October 1998 / Received revision: 29 December 1998 / Accepted: 30 December 1998 |
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