Comparative analysis of PHAs production by Bacillus megaterium OUAT 016 under submerged and solid-state fermentation.

In view of risk coupled with synthetic polymer waste, there is an imperative need to explore biodegradable polymer. On account of that, six PHAs producing bacteria were isolated from mangrove forest and affilated to the genera Bacillus & Pseudomonas from morpho-physiological characterizations. Among which the potent PHAs producer was identified as Bacillus megaterium OUAT 016 by 16S rDNA sequencing and in-silico analysis. This research addressed a comparative account on PHAs production by submerged and solid-state fermentation pertaining to different downstream processing. Here, we established higher PHAs production by solid-state fermentation through sonication and mono-solvent extraction. Using modified MSM media under optimized conditions, 49.5% & 57.7% of PHAs were produced in submerged and 34.1% & 62.0% in solid-state fermentation process. Extracted PHAs was identified as a valuable polymer PHB-co-PHV and its crystallinity & thermostability nature was validated by FTIR, 1H NMR and XRD. The melting (Tm) and thermal degradation temperature (Td) of PHB-co-PHV was 166 °C and 273 °C as depicted from DTA. Moreover, FE-SEM and SPM surface imaging indicated biodegradable nature, while FACS assay confirmed cytocompatibility of PHB-co-PHV.

Growth associated polyhydroxybutyrate production by the novel Zobellellae tiwanensis strain DD5 from banana peels under submerged fermentation.

In view of environmental pollution by fossil fuel-based plastics, it has become imperative to find out an alternative biodegradable plastic for sustainability. In this context, polyhydroxy butyrate (PHB) production was carried out by the Zobellella sp. DD5 using inexpensive banana peels as the carbon source. Under optimized condition, 1.13 g/L (47.3%) of PHB was produced by the bacteria in growth associated mechanism. The CO group of PHB was detected from the high intense absorption band (1719 cm-1) of FTIR spectroscopic analysis. NMR and GC-MS results are also identical with the chemical shift signal CH, CH2 and CH3 group of PHB. The PHB is crystalline in nature and degree of crystallinity (Xc) - 34.38%, melting temperature (Tm) - 169 °C, thermal decomposition temperature (Td) - 248 °C as detected by XRD and DTA respectively. Rough surface morphology of PHB film was validated by AFM and SEM imaging that improves biodegradability of the PHB. The Young's modulus, tensile strength and elongation at break depicted hard and brittle nature of PHB. Fluorescence-activated cell sorting (FACS) confirmed cytocompatibility of PHB at 500 µg/mL in human embryonic kidney (HEK-293) cell line. The non-cytotoxic PHB can be used for various biomedical and agricultural applications in future.

Bioconversion of fish solid waste into PHB using Bacillus subtilis based submerged fermentation process.

Currently, one of the major problem affecting the world is solid waste management, predominantly petroleum-based plastic and fish solid waste (FSW). However, it is very difficult to reduce the consumption of plastic as well as fish products, but it is promising to convert FSW to biopolymer to reduce eco-pollution. On account of that, the bioconversion of FSW extract to polyhydroxybutyrate (PHB) was undertaken by using Bacillus subtilis (KP172548). Under optimized conditions, 1.62 g/L of PHB has been produced by the bacterium. The purified compound was further characterized by advanced analytical technologies to elucidate its chemical structure. Results indicated that the biopolymer was found to be PHB, the most common homopolymer of polyhydroxyalkanoates (PHAs). This is the first report demonstrating the efficacy of B. subtilis to utilize FSW extract to produce biopolymer. The biocompatibility of the PHB against murine macrophage cell line RAW264.7 demonstrated that, it was comparatively less toxic, favourable for surface attachment and proliferation in comparison with poly-lactic acid (PLA) and commercially available PHB. Thus, further exploration is highly indispensable to use FSW extract as a substrate for production of PHB at pilot scale.

Structural and thermal characterization of PHAs produced by Lysinibacillus sp. through submerged fermentation process.

In this investigation an attempt has been made to characterize and identify Lysinibacillus sp. 3HHX by 16S-rDNA sequencing. The bacterium exhibited occurrence of PHAs granules on an average 11±1 per cell of 1.0µm length and breadth 0.72µm, revealed from TEM studies. Under optimized condition, 4.006gm/L of PHAs was extracted using hypochlorite digestion and multi-solvent extraction process. PhaC gene of ∼540bp and higher PHA synthase activity was detected at 48h of cultivation. The extracted PHAs was structurally characterized by GC-MS and 1H NMR reported to be P(3HB-co-3HDD-co-3HTD) and amorphous in nature with 112°C melting point, -11.0°C glass transition point and 114.76°C decomposition temperature detected by DSC & TGA respectively. The C/O of biopolymer disc was 1:65 as revealed from C1s and O1s spectra of XPS, that was completely biodegradable within 30 days. This biopolymer was observed to be non-cytotoxic to NIH 3T3 mouse fibroblast cells. The report is of its kind in establishing the abilities of Lysinibacillus sp. 3HHX for non-growth associated PHA co-polymer production. Moreover the biocompatible and biodegradable nature of the biopolymer conferred to its substantial biomedical applications.