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Biocompatible polyhydroxybutyrate (PHB) production by marine Vibrio azureus BTKB33 under submerged fermentation

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Abstract

Polyhydroxybutyrate (PHB) is known to have applications as medical implants and drug delivery carriers and is consequently in high demand. In the present study the possibilities of harnessing potential PHB-producing vibrios from marine sediments as a new source of PHB was investigated since marine environments are underexplored. Screening of polyhydroxyalkanoate (PHA)-producing vibrios from marine sediments was performed using a fluorescent plate assay followed by spectrophotometric analysis of liquid cultures. Out of 828 isolates, Vibrio sp. BTKB33 showed maximum PHA production of 0.21 g/L and PHA content of 193.33 mg/g of CDW. The strain was identified as Vibrio azureus based on phenotypic characterization and partial 16S rDNA sequence analysis. The strain also produced several industrial enzymes: amylase, caseinase, lipase, gelatinase, and DNase. The FTIR analysis of extracted PHA and its comparison with standard PHB indicated that the accumulated PHA is PHB. Bioprocess development studies for enhancing PHA production were carried out under submerged fermentation conditions. Optimal submerged fermentation conditions for enhanced intracellular accumulation of PHA production were found to be 35 °C, pH −7, 1.5 % NaCl concentration, agitation at 120 rpm, 12 h of inoculum age, 2.5 % initial inoculum concentration, and 36 h incubation along with supplementation of magnesium sulphate, glucose, and ammonium chloride. The PHA production after optimization was found to be increased to 0.48 g/L and PHA content to 426.88 mg/g of CDW, indicating a 2.28-fold increase in production. Results indicated that V. azureus BTKB33 has potential for industrial production of PHB.

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Acknowledgement

This study was supported by project grants from the Department of Ocean Development, Government of India (No. DOD/11 – MRDF/1/29/06) and the Centre for Marine Living Resources & Ecology, Ministry of Earth Sciences, Government of India (MOES/10-MLR/2/2007) given to Dr. Sarita G. Bhat, Dept. of Biotechnology, Cochin University of Science and Technology, Kochi, India.

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  1. Department of Biotechnology, Cochin University of Science and Technology, Kalamassery, Cochin-22, Kerala, India

    Raghul Subin Sasidharan, Sarita Ganapathy Bhat & Muthusamy Chandrasekaran

  2. Department of Botany & Microbiology, College of Science, King Saud University, PB No. 2455, Riyadh, 11451, Kingdom of Saudi Arabia

    Muthusamy Chandrasekaran

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  1. Raghul Subin Sasidharan
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  2. Sarita Ganapathy Bhat
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Correspondence to Sarita Ganapathy Bhat.

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Sasidharan, R.S., Bhat, S.G. & Chandrasekaran, M. Biocompatible polyhydroxybutyrate (PHB) production by marine Vibrio azureus BTKB33 under submerged fermentation. Ann Microbiol 65, 455–465 (2015). https://doi.org/10.1007/s13213-014-0878-z

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