Draft Genome Sequence of Bacillus aryabhattai Strain PHB10, a Poly(3-Hydroxybutyrate)-Accumulating Bacterium Isolated from Domestic Sewerage

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

Pillai

Kumar

Harikrishnan

2020

3 Biotech

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Enhanced production of poly(3-hydroxybutyrate) in recombinant Escherichia coli and EDTA–microwave-assisted cell lysis for polymer recovery

2018

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Poly(3-hydroxybutyrate) (PHB) is a bacterial polymer of great commercial importance due to its properties similar to polypropylene. With an aim to develop a recombinant system for economical polymer production, PHB biosynthesis genes from Bacillus aryabhattai PHB10 were cloned in E. coli. The recombinant cells accumulated a maximum level of 6.22 g/L biopolymer utilizing glycerol in shake flasks. The extracted polymer was confirmed as PHB by GC–MS and NMR analyses. The polymer showed melting point at 171 °C, thermal stability in a temperature range of 0–140 °C and no weight loss up to 200 °C. PHB extracted from sodium hypochlorite lysed cells had average molecular weight of 143.108 kDa, polydispersity index (PDI) 1.81, tensile strength of 14.2 MPa and an elongation at break of 7.65%. This is the first report on high level polymer accumulation in recombinant E. coli solely expressing PHB biosynthesis genes from a Bacillus sp. As an alternative to sodium hypochlorite cell lysis mediated polymer extraction, the effect of combined treatment with ethylenediaminetetraacetic acid and microwave was studied which attained 93.75% yield. The polymer recovered through this method was 97.21% pure, showed 2.9-fold improvement in molecular weight and better PDI. The procedure is simple, with minimum polymer damage and more eco-friendly than the sodium hypochlorite lysis method.

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Draft Genome Sequence of Bacillus aryabhattai Strain PHB10, a Poly(3-Hydroxybutyrate)-Accumulating Bacterium Isolated from Domestic Sewerage

Cited by 2 publications

References 7 publications

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

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

Enhanced production of poly(3-hydroxybutyrate) in recombinant Escherichia coli and EDTA–microwave-assisted cell lysis for polymer recovery

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