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Accumulation of polyhydroxyalkanoates by halophilic archaea isolated from traditional solar salterns of India

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Abstract

Extremely halophilic archaeal isolates obtained from brine and sediment samples of solar salterns of Goa and Tamil Nadu, India were screened for accumulation of polyhydroxyalkanoates (PHA). Seven polymer accumulating haloarchaeal strains (TN4, TN5, TN6, TN7, TN9, TN10 and BBK2) were selected based on their growth and intensity of fluorescence when grown on 20 % NaCl synthetic medium supplemented with 2 % glucose and incorporated with Nile red dye. The polymer was quantified by conversion of PHA to crotonic acid which gave a characteristic absorption maxima at 235 nm. On the basis of phenotypic and genotypic characterization the cultures TN4, TN5, TN6, TN7, TN10 and BBK2 were grouped under genus Haloferax whereas isolate TN9 was grouped under the genus Halogeometricum. Growth kinetics and polymer accumulation studies revealed that the culture Halogeometricum borinquense strain TN9 accumulates PHA maximally at the mid-log phase, i.e. 5th day of growth (approx. 14 wt% PHA of CDW). Analysis of the polymer by IR, 1H NMR and 13C NMR confirmed it to be a homopolymer of 3-hydroxybutyrate.

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Abbreviations

PHA:

Polyhydroxyalkanoates

PHB:

Polyhydroxybutyrate

NTYE:

NaCl tryptone yeast extract

NT:

NaCl tri-sodium citrate

NSM:

NaCl synthetic media

DMSO:

Dimethylsulfoxide

PI:

Propidium iodide

w/v:

Weight by volume

EPS:

Exopolysaccharide

CDW:

Cell dry weight

rpm:

Revolutions per minute

IR:

Infra red

NMR:

Nuclear magnetic resonance

TGA:

Thermo gravimetric analysis

DTA:

Differential thermal analysis

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Acknowledgments

This work was supported by University Grants Commission, India (UGC) Major Research Project No: 34–500/2008(SR). BBS and KM would like to thank Council of Scientific and Industrial Research India for awarding Senior Research Fellowship (SRF) (09/919(0016)/2012-EMR-I and (09/919(0017)/2012-EMR-I). Authors are grateful to Dr. Narendra Nath Ghosh Department of Chemistry for helping with TGA–DTA analysis and Mr. Areef Sardar, NIO Goa for SEM analysis.

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Authors and Affiliations

  1. Department of Biological Sciences, Birla Institute of Technology and Science Pilani, K K Birla Goa Campus, NH 17 B, Zuarinagar, 403 726, Goa, India

    Bhakti B. Salgaonkar, Kabilan Mani & Judith Maria Bragança

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  1. Bhakti B. Salgaonkar
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  2. Kabilan Mani
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  3. Judith Maria Bragança
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Corresponding author

Correspondence to Judith Maria Bragança.

Additional information

Communicated by F. Robb

Electronic supplementary material

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792_2013_561_MOESM1_ESM.tif

Supplementary Fig. S1 Haloarchaeal isolates (BK20, BBK2, TN9 and BS2) grown on solid NTYE and NT media containing 25 % (w/v) NaCl (TIFF 1701 kb)

792_2013_561_MOESM2_ESM.tif

Supplementary Fig. S2 SEM micrographs of the Haloarchaeal isolates (a) TN5, (b) TN9, (c) BK20 and (d) BS16. (TIFF 1319 kb)

792_2013_561_MOESM3_ESM.tif

Supplementary Fig. S3 Extremely haloarchaeal cultures (TN5 and TN9) grown in 20 % NaCl synthetic medium (NSM) and 2 % glucose. (TIFF 2839 kb)

792_2013_561_MOESM4_ESM.tif

Supplementary Fig. S4 Spectrophotometric scans of Crotonic Acid of acid digested PHA obtained from haloarchaeal Isolate TN9, BBK2 and standard PHB (Sigma-Aldrich). (TIFF 212 kb)

Supplementary Fig. S5 Infrared spectrum of polymer obtained from TN9. (TIFF 182 kb)

Supplementary Fig. S6 1H NMR spectrum of polymer of TN9 (TIFF 140 kb)

Supplementary Fig. S7 13C NMR spectrum of polymer of TN9. (TIFF 125 kb)

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Salgaonkar, B.B., Mani, K. & Bragança, J.M. Accumulation of polyhydroxyalkanoates by halophilic archaea isolated from traditional solar salterns of India. Extremophiles 17, 787–795 (2013). https://doi.org/10.1007/s00792-013-0561-5

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