Izvestiya of Saratov University.

Chemistry. Biology. Ecology

ISSN 1816-9775 (Print)
ISSN 2541-8971 (Online)

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Fourier Transform Infrared Spectroscopic Analysis of Poly-3-Hydroxybutyrate Accumulation by Azospirillum brasilense Cells at Various Cultivation Periods and Ammonium Concentrations in the Culture Medium

Parshina Victoria V., Saratov State University
Dyatlova Yulia A., Institute of Biochemistry and Physiology of Plants and Microorganisms of the Russian Academy of Sciences
Tugarova Anna V., Institute of Biochemistry and Physiology of Plants and Microorganisms of the Russian Academy of Sciences

Many bacteria, in response to unfavourable environmental conditions, can synthesise polyesters of the polyhydroxyalkanoate (PHA) class. These biopolymers, accumulating intracellularly in the form of granules, help the bacteria to cope with negative environments and are utilised as reserve sources of carbon and energy. Rhizobacteria of the species Azospirillum brasilense synthesise a single type of PHA, poly-3-hydroxybutyrate (PHB), in response to stress factors. Knowledge of the principles and conditions of PHB synthesis is of importance both for understanding the subsistence of azospirilla in their natural habitats and for their use as biofertilisers, to preserve their viability. In this work, Fourier transform infrared (FTIR) spectroscopy was used to study PHB accumulation by cells of Azospirillum brasilense strains Sp7 и Sp245 during growth of the bacteria for 6 days in standard malate salt medium containing various concentrations of ammonium chloride (0.05, 0.10 and 0.21 g/l). Comparative analysis of FTIR spectra of the bacterial biomass samples showed that, at suboptimal initial concentrations of bound nitrogen in the medium, PHB accumulation after 1 to 6 days of cultivation was more intensive in A. brasilense strain Sp7 than in strain Sp245. Maximum relative amounts of PHB were accumulated by biomass of A. brasilense Sp7 grown for 3 days at the initial NH4Cl concentration in the culture medium 0.10 g/l.


1. Kadouri D., Jurkevitch E., Okon Y., Castro-Sowinski S. Ecological and agricultural significance of bacterial polyhydroxyalkanoates // Crit. Rev. Microbiol. 2005. Vol. 31. P. 55–67.

2. Bashan Y., de-Bashan L. E. How the plant growthpromoting bacterium Azospirillum promotes plant growth – a critical assessment // Adv. Agron. 2010. Vol. 108. P. 77–136.

3. Kamnev A. A., Tugarova A. V., Antonyuk L. P., Tarantilis P. A., Kulikov L. A., Perfi liev Yu. D., Polissiou M. G., Gardiner P. H. E. Instrumental analysis of bacterial cells using vibrational and emission Mossbauer spectroscopic techniques // Anal. Chim. Acta. 2006. Vol. 573–574. P. 445–452.

4. Kamnev A. A. FTIR spectroscopic studies of bacterial cellular responses to environmental factors, plantbacterial interactions and signalling // Spectrosc. Intern. J. 2008. Vol. 22, № 2–3. P. 83–95.

5. Kamnev A. A., Sadovnikova J. N., Tarantilis P. A., Polissiou M. G., Antonyuk L. P. Responses of Azospirillum brasilense to nitrogen defi ciency and to wheat lectin : a diffuse refl ectance infrared Fourier transform (DRIFT) spectroscopic study // Microb. Ecol. 2008. Vol. 56, № 4. P. 615–624.

6. Kamnev A. A., Tugarova A. V., Tarantilis P. A., Gardiner P. H. E., Polissiou M. G. Comparing poly-3- hydroxybutyrate accumulation in Azospirillum brasilense strains Sp7 and Sp245 : the effects of copper(II) // Appl. Soil Ecol. 2012. Vol. 61. P. 213–216.

7. Fibach-Paldi S., Burdman S., Okon Y. Key physiological properties contributing to rhizosphere adaptation and plant growth promotion abilities of Azospirillum brasilense // FEMS Microbiol. Lett. 2011. Vol. 326, № 2. P. 99–108.

8. Cassan F., Diaz-Zorita M. Azospirillum sp. in current agriculture : From the laboratory to the fi eld // Soil Biol. Biochem. 2016. Vol. 103. P. 117–130.

9. Naumann D. Infrared Spectroscopy in Microbiology // Encyclopedia of Analytical Chemistry / ed. R. A. Meyers. Chichester : Wiley, 2000. P. 102–131.

10. Kamnev A. A., Mamchenkova P. V., Dyatlova Yu. A., Tugarova A. V. FTIR spectroscopic studies of selenite reduction by cells of the rhizobacterium Azospirillum brasilense Sp7 and the formation of selenium nanoparticles // J. Mol. Struct. 2017. Vol. 1140. P. 106–112.

11. Tugarova A. V., Shelud’ko A. V., Dyatlova Yu. A., Filip’echeva Yu. A., Kamnev A. A. FTIR spectroscopic study of biofi lms formed by the rhizobacterium Azospirillum brasilense Sp245 and its mutant Azospirillum brasilense Sp245.1610 // J. Mol. Struct. 2017. Vol. 1140. P. 142–147.

12. Tugarova A. V., Mamchenkova P. V., Dyatlova Yu. A., Kamnev A. A. FTIR and Raman spectroscopic studies of selenium nanoparticles synthesised by the bacterium Azospirillum thiophilum // Spectrochim. Acta Part A : Mol. Biomol. Spectrosc. 2018. Vol. 192. P. 458–463.

13. Kamnev A. A., Tugarova A. V., Dyatlova Yu. A., Tarantilis P. A., Grigoryeva O. P., Fainleib A. M., De Luca S. Methodological effects in Fourier transform infrared (FTIR) spectroscopy : Implications for structural analyses of biomacromolecular samples // Spectrochim. Acta Part A : Mol. Biomol. Spectrosc. 2018. Vol. 193. P. 558–564.

14. Tarrand J. J., Krieg N. R., Dobereiner J. A taxonomic study of the Spirillum lipoferum group, with descriptions of a new genus, Azospirillum gen. nov. and two species, Azospirillum lipoferum (Beijerinck) comb. nov. and Azospirillum brasilense sp. nov // Can. J. Microbiol. 1978. Vol. 24, №. 8. P. 967–980.

15. Baldani V. L. D., Baldani J. I., Dobereiner J. Effects of Azospirillum inoculation on root infection and nitrogen incorporation in wheat // Can. J. Microbiol. 1983. Vol. 29, №. 8. P. 924–929.

16. Day J. M., Dobereiner J. Physiological aspects of N2- fi xation by a Spirillum from Digitaria roots // Soil Biol. Biochem. 1976. Vol. 8, № 1. P. 45–50.

17. Kansiz M., Billman-Jacobe H., McNaughton D. Quantitative determination of the biodegradable polymer poly(?hydroxybutyrate) in a recombinant Escherichia coli strain by use of mid-infrared spectroscopy and multivariative statistics // Appl. Envir. Microbiol. 2000. Vol. 66, № 8. P. 3415–3420.

18. Kansiz M., Dominguez-Vidal A., McNaughton D., Lendl B. Fourier-transform infrared (FTIR) spectroscopy for monitoring and determining the degree of crystallisation of polyhydroxyalkanoates (PHAs) // Anal. Bioanal. Chem. 2007. Vol. 388, № 5–6. P. 1207–1213.