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Biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) in Bacillus aryabhattai and cytotoxicity evaluation of PHBV/poly(ethylene glycol) blends

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The study described poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) accumulation in Bacillus aryabhattai PHB10 for the first time and evaluated the polymer induced cytotoxicity in-vitro with PHBV/poly(ethylene glycol) (PEG) blends. The B. aryabhattai strain produced 2.8 g/L PHBV, equivalent to 71.15% of cell dry mass in a medium supplemented with propionic acid, after 48 h incubation. The optimum temperature and pH for the copolymer accumulation was 31 °C and 7, respectively. The gas chromatography–mass spectrometry and nuclear magnetic resonance analyses confirmed the polymer obtained as PHBV. The differential scanning calorimetry analysis revealed that the melting point of the material as 90 °C and its thermal stability up to 220 °C. The average molecular weight (Mn) and polydispersity index of the sample was estimated by gel permeation chromatography analysis and observed as 128.508 kDa and 2.82, respectively. The PHBV showed tensile strength of 10.3 MPa and elongation at break of 13.3%. The PHBV and their blends with PEG were tested for cytotoxicity on human keratinocytes (HaCaT cells) and the cells incubated with PHBV/PEG2kDa blends were 99% viable, whereas with the PHBV alone showed comparatively higher cytotoxicity. The significant improvement in the cell viability of PHBV/PEG2kDa blends indicates its potential as a candidate for skin graft applications.

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The authors are grateful to Prof. M. Radhakrishna Pillai, Director, RGCB for providing the facilities; the Kerala Forest Research Institute, Peechi for GC–MS analysis and the SAIF at STIC-Kochi for the NMR and TGA facilities.


This work was supported by funding from the Department of Biotechnology (DBT), Government of India as an intramural grant. ABP received a research fellowship from the Council of Scientific and Industrial Research (CSIR), India (SRF; 09/716(0149)/2012-EMR-I).

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ABP and HK conceived and designed the experiments; ABP and AJK did the experiments; ABP, AJK and HK analyzed the data and wrote this manuscript. All authors read and approved the final manuscript.

Correspondence to Harikrishnan Kumarapillai.

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Balakrishna Pillai, A., Jaya Kumar, A. & Kumarapillai, H. Biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) in Bacillus aryabhattai and cytotoxicity evaluation of PHBV/poly(ethylene glycol) blends. 3 Biotech 10, 32 (2020).

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  • Bacillus aryabhattai PHB10
  • Cytotoxicity
  • Gas chromatography–mass spectrometry
  • HaCaT cells
  • Poly(ethylene glycol)
  • Poly(3-hydroxybutyrate-co-3-hydroxyvalerate)