Characterization of polyhydroxyalkanoates accumulated by a moderately halophilic salt pan isolate Bacillus megaterium strain H16
Article first published online: 13 FEB 2013
© 2013 The Society for Applied Microbiology
Journal of Applied Microbiology
Volume 114, Issue 5, pages 1347–1356, May 2013
How to Cite
Salgaonkar, B.B., Mani, K. and Braganca, J.M. (2013), Characterization of polyhydroxyalkanoates accumulated by a moderately halophilic salt pan isolate Bacillus megaterium strain H16. Journal of Applied Microbiology, 114: 1347–1356. doi: 10.1111/jam.12135
- Issue published online: 15 APR 2013
- Article first published online: 13 FEB 2013
- Accepted manuscript online: 13 JAN 2013 02:20AM EST
- Manuscript Accepted: 2 JAN 2013
- Manuscript Revised: 21 DEC 2012
- Manuscript Received: 10 SEP 2012
- University Grants Commission, India (UGC). Grant Number: 34-500/2008SR
- Bacillus megaterium strain H16;
- polymer characterization;
Characterization of polyhydroxyalkanoates (PHA) accumulated by halophilic bacteria isolated from solar salterns.
Methods and Results
Twenty-six halophilic isolates were obtained from solar salterns of Goa, India. They were screened for accumulation of PHA by Sudan black B, Nile blue A and Nile red stains. Strains H15, H16 and H26 were selected based on their intensity of Nile blue A/Nile red fluorescence. On the basis of phenotypic and genotypic characterization, the three isolates were identified as Bacillus megaterium. Growth kinetics and polymer accumulating capacity of strain H16 were studied in E2 mineral media with 2% glucose with/without NaCl. In the absence of NaCl, strain H16 accumulated PHA to 40·0% (w/w) of cell dry weight (CDW) at 42 h of growth, whereas in presence of 5% w/v NaCl, the culture showed longer lag phase of up to 24 h and accumulated a maximum PHA of 39% (w/w) CDW at 54 h of growth. The infrared spectra of both the polymers exhibited peaks at 1733·9 cm−1 characteristic of C=O. Scans of 1H nuclear magnetic resonance (NMR) showed a doublet at 2·5 ppm corresponding to methylene group (-CH2), the signal at 5·3 ppm corresponded to methine group (-CH-), and another signal at 1·3 ppm corresponded to the methyl group (-CH3). Scans of 13C NMR showed prominent peaks at 20, 40, 67–68 and 170 ppm, indicating the polymer to be homopolymer of 3-hydroxybutyrates. The polymer is stable up to a temperature of 160°C.
Three moderately halophilic isolates (strain H15, H16 and H26) capable of accumulating PHA were isolated from solar salterns of Ribandar Goa, India, and identified as B. megaterium based on phenotypic and genotypic characterization. Strain H16 accumulated polyhydroxybutyrate in the presence and absence of NaCl up to 40% of its CDW.
Significance and Impact of the Study
This strain would be better suited for production of PHA at industrial level due to its tolerance to high concentration of NaCl.