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Drotaverine Hydrochloride Degradation Using Cyst-like Dormant Cells of Rhodococcus ruber

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Abstract

This work has a focus on adaptive capabilities of the actinobacterium Rhodococcus ruber IEGM 326 to cope with drotaverine hydrochloride (DH), a known pharmaceutical pollutant. Cultivation of R. ruber in a nitrogen-limited medium with incubation at the ambient temperature resulted in the formation of cyst-like dormant cells (CLDCs). They maintained viability for 2–7 months, possessed the undetectable respiratory activity and elevated resistance to heating, and had a specific morphology. CLDCs are regarded to ensure long-term survival in various habitats and may be used as storage formulations. R. ruber IEGM 326 was tolerant to DH (MIC, 200 mg/l) and displayed different abilities to degrade this compound, depending on inoculum, temperature, and the presence of glucose as co-oxidized substrate. Thus, the loss of DH (20 mg/l) over 48 h at the optimal temperature (27 ± 2 °C) was 5–8 % in the absence of glucose after inoculating with vegetative cells. The addition of glucose (5 g/l) increased DH degradation up to 46 %. Noteworthy, CLDCs as inoculum were advantageous over vegetative cells to degrade DH at the non-optimal temperature (35 ± 2 °C) at reduced bulk respiratory activity. The obtained results are promising to improve the biodegrading capabilities of other Rhodococcus strains.

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Acknowledgments

The research was partially supported by the Grant of the Russian Academy of Sciences Presidium Program “Leaving Nature: Current State and Development Problems” (01201256869) and the Russian Scientific Foundation Grant (14-14-00643).

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Authors and Affiliations

  1. Institute of Ecology and Genetics of Microorganisms, Russian Academy of Sciences, 13 Golev Street, 614081, Perm, Russia

    Irena B. Ivshina & Anna N. Mukhutdinova

  2. Microbiology and Immunology Department, Perm State National Research University, Perm, Russia

    Irena B. Ivshina & Helena A. Tyumina

  3. Perm State Pharmaceutical Academy, Perm, Russia

    Helena V. Vikhareva

  4. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Pushchino, Moscow, Russia

    Nataliya E. Suzina

  5. Winogradsky Institute of Microbiology, Russian Academy of Sciences, Moscow, Russia

    Galina I. El’-Registan & Andrey L. Mulyukin

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  1. Irena B. Ivshina
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  2. Anna N. Mukhutdinova
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  4. Helena V. Vikhareva
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Correspondence to Irena B. Ivshina.

Electronic supplementary material

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284_2014_718_MOESM1_ESM.pdf

Online Resource 1 Morphology of Rhodococcus ruber IEGM 326. Vegetative cells and CLDCs in a phase contrast microscope (a) and TEM images of CLDCs (c, d) (SR1 medium, N-limited, 4-month-old cultures). Designations: CW, cell wall; CL, capsular layer; I, electron-transparent inclusions. Scale bars: 1 μm for micrographs of cell sections and 2 μm for light microscopy images

284_2014_718_MOESM2_ESM.pdf

Online Resource 2 Thermal resistance (% of the control) of R. ruber IEGM 326: vegetative cells (1) and CLDCs at different ages: 2 months (2) and 7 months (3) to heating at 55–90 °C for 10 min

284_2014_718_MOESM3_ESM.pdf

Online Resource 3 Respiration rate (a) and CO2 release dynamics (b) during DH biodegradation as the sole carbon and energy source in the medium inoculated with vegetative cell (3) and CLDC (2) suspensions of R. ruber IEGM 326. Cultures were incubated at 35 °C. Un-inoculated (abiotic) controls (1)

284_2014_718_MOESM4_ESM.pdf

Online Resource 4 Respiration rate (a) and CO2 release dynamics (b) during DH biodegradation in glucose-supplemented medium following inoculations with vegetative cell (3) and CLDC (2) suspensions of R. ruber IEGM 326. Un-inoculated (abiotic) controls (1). Cultures were incubated at 27 °C

284_2014_718_MOESM5_ESM.pdf

Online Resource 5 Chromatographic separation of DH metabolites produced by R. ruber IEGM 326 vegetative cells. Mass spectra of DH (m/z = 411.2) as well as of protocatechuic acid derivates (m/z = 137.0; 193.0; 109.0; 239.84) accumulated in the RS medium. The same product was identified when CLDCs were used as inocula (spectra are not shown here)

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Ivshina, I.B., Mukhutdinova, A.N., Tyumina, H.A. et al. Drotaverine Hydrochloride Degradation Using Cyst-like Dormant Cells of Rhodococcus ruber . Curr Microbiol 70, 307–314 (2015). https://doi.org/10.1007/s00284-014-0718-1

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