Abstract
Acidiphilium cryptum is an acidophilic, heterotrophic, and metallotolerant bacteria able to use dissolved oxygen or Fe(III) as an electron sink. The ability of this extremophile to accumulate poly(3-hydroxybutyrate) (PHB) and secrete extracellular polymeric substances (EPS) has also been reported. Hence, the aim of this work is to characterize the production of PHB and EPS by the wild strain DSM2389 using glycerol in shaken flasks and bioreactor. Results showed that maximum PHB accumulation (37–42% w/w) was obtained using glycerol concentrations of 9 and 15 g L−1, where maximum dry cell weight titers reached 3.6 and 3.9 g L−1, respectively. The culture in the bioreactor showed that PHB accumulation takes place under oxygen limitation, while the redox potential of the culture medium could be used for online monitoring of the PHB production. Recovered EPS was analyzed by Fourier-transform infrared spectroscopy and subjected to gas chromatography—mass spectrometry after cleavage and derivatization steps. These analyses showed the presence of sugars which were identified as mannose, rhamnose and glucose, in a proportion near to 3.2:2.3:1, respectively. Since glycerol had not been used in previous works, these findings suggest the potential of A. cryptum to produce biopolymers from this compound at a large scale with a low risk of microbial contamination due to the low pH of the fermentation process.
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Data availability
All data generated or analyzed during this study are included in this published article and the supplementary information file.
Abbreviations
- BDM:
-
Basal defined medium
- BEM:
-
Basal E medium
- BSTFA:
-
N,O-bis(trimethylsilyl)trifluoroacetamide
- DTGS:
-
Deuterated triglycine sulfate
- DSMZ:
-
German Collection of Microorganisms and Cell Cultures
- EPS:
-
Extracellular polymeric substances
- FTIR:
-
Fourier-transform infrared spectroscopy
- GC:
-
Gas chromatography
- GMP:
-
Good manufacturing practice
- HPLC:
-
High-performance liquid chromatography
- MEM:
-
Minimum essential medium
- MS:
-
Mass spectrometry
- OD600 :
-
Optical density at 600 nm
- pE:
-
Negative logarithm of the electron activity
- PHA:
-
Polyhydroxyalkanoate
- PHB:
-
Poly(3-hydroxybutyrate)
- RI:
-
Refractive index
- rpm:
-
Revolutions per minute
- TIC:
-
Total ion current
- TMCS:
-
Trimethylchlorosilane
- TMS:
-
Trimethylsilylation
- UDP:
-
Uridine diphosphate
- UV–Vis:
-
Ultraviolet–visible
- vvm:
-
Volume of gas (under standard conditions) per volume of liquid per minute
- v/v:
-
Volume by volume
- w/w:
-
Weight by weight
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This study was funded by the National Agency for Research and Development (ANID), Chile, grants Fondecyt Iniciación 11180688 and Fondecyt Regular 1211434.
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EG, AD conceived and designed research. CZ, GZ, NM performed microbial cultures in shaken flasks and bioreactor. JPS performed EPS analysis by FTIR. MVR and AR performed the identification and quantification of glycosyl residues. EG, MVR and BP analyzed data. EG, FS and AD wrote the manuscript. All authors reviewed the manuscript.
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González, E., Zuleta, C., Zamora, G. et al. Production of poly (3-hydroxybutyrate) and extracellular polymeric substances from glycerol by the acidophile Acidiphilium cryptum. Extremophiles 27, 30 (2023). https://doi.org/10.1007/s00792-023-01313-3
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DOI: https://doi.org/10.1007/s00792-023-01313-3