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Selection of poly(R)-3-hydroxybutyric acid utilising bacteria by enrichment,optimisation and compatibility testing for consortia development
Authors:Shikha Raghuwanshi  Tithi Agarwal  Amit Yadav  MGH Zaidi  Yogesh Shouche  Reeta Goel
Institution:1. Department of Microbiology, C.B.S.H., G. B. Pant University of Agriculture and Technology, Pantnagar, Uttarakhand, India;2. Department of Chemistry, C.B.S.H., G. B. Pant University of Agriculture and Technology, Pantnagar, Uttarakhand, India;3. Microbial Culture Collection, National Centre for Cell Sciences, Pune University Campus, Ganeshkhind, Pune, India
Abstract:Bacterial strains with poly-3-hydroxybutyrate (PHB) degradation potential were isolated from waste yard soil samples of selected industrial sites in Uttarakhand, India, and two microbial consortia were developed, i.e. Consortium I comprises Pseudomonas sp. strain Rb10, Pseudomonas sp. strain Rb11 and Bacillus sp. strain Rb18, and Consortium II is composed of Lysinibacillus sp. strain Rb1, Pseudomonas sp. strain Rb13 and Pseudomonas sp. strain Rb19. The current study involved enrichment selection via liquid and semi-solid media, followed by isolation and screening of bacterial strains using PHB pellets and films. Furthermore, the identification and characterisation was done by triphasic approach. The utilisation of PHB by the characterised strains was confirmed by Fourier transform infrared spectroscopy and scanning electron microscopy. Moreover, the minimum inhibitory concentration of solubilised PHB was found to be 2.5?mg/mL, which was detected through ‘clear zone assay’. Further, the selection and biocompatibility testing of potential isolates were performed for the formation of bacterial consortia. Thus, the present work would provide direct and standardised protocol for screening and selection of potential microbiomes for biodegradation of polymers by overcoming the negative effect of organic solvents. Moreover, indigenously developed consortia would be evaluated for their in situ biodegradation potential against various bioplastic films.
Keywords:Enrichment selection  bioplastic degradation  microbial consortia  characterisation  triphasic approach  Fourier transform infrared spectroscopy  scanning electron microscopy and poly(R)-3-hydroxybutyric acid
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