Biosynthesis and characterization of PHB from Bacillus thuringiensis GVP isolated from soil

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INTRODUCTION Today synthetic polymer has become essential part of our life. Petrochemical materials are used to synthetic polymer and plastic that are defiant to biodegradation which poses a serious threat to environment. Environment issue due to pollution and increase in the price of petroleum have made us to develop bioplastic as a replacement for petrochemical derived plastic.(Abhishek dutt tripathi)(Gulab singh)(Nisha V.Ramadas) Poly (3-hydroxybutyrate) is a widely studied biopolyhydroxyester belongs to polyhydroxyalkanoates (PHAs) family(jose Geraldo da cruz).PHB has become substitute for petrochemical derived plastic due to similar mechanical properties Poly (3-hydroxybutyrate) is ecofriendly, biodegradable, biocompatible and microbial thermoplastic(Vishnuvardhan Reddy). PHB is a straight chain homopolymer of D (–)-β -hydroxybutyric acid and are accumulated as intracellular granules in several bacterial species. Biosynthesis of PHB begin with condensation of two molecules of acetyl-CoA yield acetoacetyl-CoA, by coupling with the oxidation of NADH to NAD+ acetoacetyl-CoA consequently reduced to hydroxylbutyryl- CoA. In the existence of PHA synthase (phac) hydroxylbutyryl- CoA is polymerised into PHB, synthesized occur during exponential growth phase and transferred into important reserves. PHB is synthesized under nutrient imbalance such as limiting the concentration of essential nutrients like nitrogen or phosphorus while carbon is used in excess as energy source.Alcaligenes, Pseudomonas, Bacillus, Rhodococcus, Staphylococcus and micrococcus are some bacterial species which naturally produces PHB (Nicola´s D. Ayub Aarthi.N,Alejandro Lopez-Corte). PHB from these Gram-negative organisms is widely used in variety of pro... ... middle of paper ... ...t different pH values (5,6,7 and 8). Each condition was maintained in different flask and PHB production was determined. Effect of various carbon substrates Effect of carbon source was determined by replacing glucose with agro-industrial waste like molasses, whey and spent wash. PHB content in the cell was determined as described earlier. Characterization of PHB FTIR spectroscopy Extracted Polymer was thoroughly mixed with KBr. KBr pellet was dried at 100°C for 4 hr. Analysis was done by Fourier Transform IR Spectrometer (Perkin-Elmer,USA) by placing the sample on the holder. The scanning conditions were a spectra range of 4000-400 cm-1. 1H-NMR spectroscopy The 1H-NMR of the precipitate was carried out using BRUKER SPEC Instrument, Germany. The polymer was dissolved in CDCL3 and vortexed for 10 min, sample was placed into NMR tubes and then analysed.

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