Abstract
Vinasse, a recalcitrant waste of the ethanol industry was employed for the production of polyhydroxyalkanoate (PHA) by the extremely halophilic archaeon, Haloarcula marismortui in shake flasks. The PHA was recovered by osmotic lysis of the cells and subsequent purification by sodium hypochlorite and organic solvents. Through UV–vis spectroscopy, differential scanning calorimetry, Fourier transform infrared, and nuclear magnetic resonance spectroscopy, the PHA was found to have characteristics very similar to that of the standard polyhydroxybutyrate (PHB) from Sigma. Inhibitory effect of polyphenols contained in vinasse was assessed by a quick and reliable cup-plate agar-diffusion method. Raw vinasse (10%) was utilized leading to accumulation of 23% PHA (of cell dry weight) and following an efficacious pre-treatment process through adsorption on activated carbon, 100% pre-treated vinasse could be utilized leading to 30% accumulation of PHB by H. marismortui. Maximum specific growth rate, specific production rate, and volumetric productivity attained using 10% raw vinasse were comparable to that obtained using a previously reported nutrient deficient medium (NDM), while the values with 100% pre-treated vinasse were higher than that determined using NDM medium. This is the first report of polyhydroxybutyrate production by a halophilic microorganism utilizing vinasse.
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Acknowledgements
Authors wish to thank Mr. Swapan K Bayen, Mr. Santanu Ghose, and Mr. Anindya Roy of IFB Agro Industries for the gift of the vinasse. Assistance from Mr. Biswarup Saha during fluorescence microscopy is thankfully acknowledged.
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Supplementary Figure 1
UV–vis scanning spectroscopy following hydrolysis and dehydrogenation of standard PHB from Sigma (red line) and PHA from H. marismortui (blue line) (JPEG 88 kb)
Supplementary Figure 2
DSC thermograms of a PHA from H. marismortui (JPEG 74 kb)
Supplementary Figure 2
DSC thermograms of b standard PHB from Sigma (JPEG 71 kb)
Supplementary Figure 3
FTIR spectra of PHA from H. marismortui (lower trace), compared with standard PHB from Sigma (upper trace) (JPEG 265 kb)
Supplementary Figure 4
1H NMR spectra of a PHA from H. marismortui (JPEG 204 kb)
Supplementary Figure 4
1H NMR spectra of b standard PHB from Sigma (JPEG 134 kb)
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Pramanik, A., Mitra, A., Arumugam, M. et al. Utilization of vinasse for the production of polyhydroxybutyrate by Haloarcula marismortui . Folia Microbiol 57, 71–79 (2012). https://doi.org/10.1007/s12223-011-0092-3
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DOI: https://doi.org/10.1007/s12223-011-0092-3