Recovery and characterization of poly(3-Hydroxybutyric acid) synthesized in Staphylococcus epidermidis

Abstract

Polyhydroxyalkanoates (PHA) are biodegradable polyesters accumulated intracellularly as energy resources by bacterial species. In this study, fermentation process for production of PHA is carried out using sesame oil as carbon source. We studied recovery of poly(3-hydroxybutyric acid) (PHB) from Staphylococcus epidermidis by sodium hypochlorite digestion method. Recovered PHB sample was estimated by UV spectrophotometer. PHB from S. epidermidis was characterized and by these findings, we examined purified PHB by differential scanning calorimeter (DSC), a thermo gravimetric analyzer (TGA), thin layer chromatography (TLC) and infrared spectroscopy (IR). The results of our analysis of PHB while comparing with commercial source suggest that in DSC melting temperature of PHB was 173.36°C, TGA thermo grams of PHB sample was at 296.91°C, on TLC plate; Rf value was calculated as 0.71 and finally IR spectrum of the compounds showed characteristics bands for the groups CH, C=O and CO , indicating the presence of PHB in the production medium.

Insights into the Polyhydroxybutyrate Biosynthesis in Ralstonia solanacearum Using Parallel 13C Tracers and Comparative Genome Analysis

“…Extracted P­(3HB) along with different standard P­(3HB) concentrations (1.2, 2, 6, 8, and 10 mg mL –1 ) was dissolved in 10 mL of concentrated H 2 SO 4 and heated for 20 min at 100 °C in a water bath. P­(3HB) gets converted into crotonic acid by H 2 SO 4 reaction . The resultant crotonic acid solution was subjected to UV spectrophotometry, and the absorbance was measured at 235 nm using sulfuric acid as a blank.…”

“…P(3HB) gets converted into crotonic acid by H 2 SO 4 reaction. 32 The resultant crotonic acid solution was subjected to UV spectrophotometry, and the absorbance was measured at 235 nm using sulfuric acid as a blank. The concentration of isolated P(3HB) was calculated from the standard curve of pure P(3HB) based on absorbance Analysis of Extracted P(3HB) Using Fluorescence Microscopy, 1 H NMR, and GC-MS. To visualize the P(3HB) granules in fluorescence microscopy, bacterial cells were pelleted down by centrifugation (5000 rpm, 10 min), washed, and suspended in 1 mL of PBS and then subjected to nile red staining (1 μg/mL in acetone).…”

Analysis, Optimization and Molecular Characterization of PHB Positive Bacteria Isolated from Agricultural Soil Sample

“…10 mg of produced PHB is dissolved in 5ml of sulfuric acid and heated in a water bath for 10 minutes. The resultant brown coloured crotonic acid solution was measured for absorbance at 235nm against sulfuric acid blank in UV spectrophotometer (Darshan Marjadi et al, 2014). Quantitative test for the produced PHB was done using UV spectrophotometer (Aslim et al, 1998).…”

Poly-hydroxybutyrate (PHB) production by Halomonas elongata BK AG 18 indigenous from salty mud crater at central Java Indonesia

“…The thermal properties of PHB samples were investigated by DSC-TGA based on the peaks (endothermic curve) or valleys (exothermic curve) shown in the thermogram. The standard PHB has a melting point peak at 172.4 °C (Marjadi and Dharaiya, 2014) but in another study, it was 168.05 °C (Güngörmedi et al, 2014). Therefore, referring to the melting point of the commercial PHB, an endothermic peak at 166.5 °C is more likely the melting point of PHB.…”