Isolation of PHB Producing Rhizobacteria and Molecular Characterization of Gene(s) for PHB Synthase for Efficient Production of Polyhydroxybutyrate
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Date
2015
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CCSHAU
Abstract
Polyhydroxybutyrate the bioplastic, producing bacteria expeditiously were isolated from fifty diverse
locations and 45 of them were found PHB producing on the basis of Nile blue A colony screening
method. Among PHB producing isolates, 6 isolates were further selected on the basis of maximum
florescence on different carbon and nitrogen sources and these were evaluated for relative PHB
production. Three bacterial isolates, B2, B3 and C6 produced maximum PHB by weight and crotonic
acid production. Cultural, morphological and biochemical characterization identified them as
Pseudomonas spp. After PHB production kinetics studies, strain B3 and B2 gave maximum PHB in
three and four days respectively. Culture conditions were optimized using Response Surface
Methodology for maximum PHB production. The strain B 2 gave 37mg/l PHB and B3 gave 45mg/l of
PHB. Hence, B3 strain was selected for further studies. FTIR confirmed the properties of PHB in B3
strain. A gene phaC was PCR amplified, cloned and sequenced. Sequence analysis employing
bioinformatics tools like BLAST, PHYRE2 evaluated the predicted models. Ecofriendly and renewable
agriproducts based PHB production method developed in these investigations may in future proceed in
the direction of adopting this at commercial scale towards making PHB the agroplastic.
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Keywords
productivity, bacteria, acidity, carbon, extraction, nitrogen, mustard, glucose, proteins, yeasts