Polyhydroxyalkanoates (PHAs) have gained much attention as biodegradable polymers, many efforts are being made to minimize the cost of PHAs by finding cheap carbon source depending on the type of microorganism and fermentation conditions. The aims of this study were to evaluate the effects of different glucose concentrations and other important conditions on the PHA production by Bacillus cereus isolated from soil. Polyhydroxyalkanoates PHAs accumulated by soil microorganisms were examined by screening the isolated bacteria using Sudan B Black and Nile Blue staining process. A Gram positive strain was identified using the 16s rRNA gene, deposited in the NCBI GenBank sequence database. Different growth conditions (favorite glucose concentrations 1-8 % (w/v), temperatures and pH) were tested and the growth parameters (sugar consumption, cell counting and Cell Dry Weight CDW) were studied. The extracted polymers were analyzed and characterized using an FTIR spectrophotometer followed by a GC-MS analysis. The pure bacterial strain isolated from soil was deposited in the NCBI GenBank database B. cereus strain ARY73, which showed significant black colored granules (or dark blue) using Sudan B Black stain, it also showed positive to Nile blue A as a high indicator stain for PHA accumulation. B. cereus ARY73 showed high production of PHA using (w/v): 2% glucose and 1% nitrogen source at 35 °C and pH7 yields 79% per Cell Dry Weight and 96 h of incubation. The extracted polymers were analyzed and characterized using an FTIR spectrophotometer confirming the PHA structure. The FTIR spectrophotometer, followed by a GC-MS analysis indicated the Scl-co-mcl PHA structure. This research demonstrates that the isolated strain B. cereus ARY73 was a good candidate for PHA production with a better quality for use in biomedical and other applications. The use of biopolymer in soil, enhanced the accumulation of the microorganisms (such as bacteria) capable of degrading biopolymer or biodegradation by-products yields by other species which were isolated in this study and demonstrated their efficiency in producing biopolymers.
The production of polyhydroxyalkanoates PHAs from biopolymer degrading bacteria was examined in situ by screening isolates using Sudan B Black staining process as potential PHAs detecting, and Nile Blue staining as a proof method detection. Five bacterial strains isolated from biopolymer waste buried in a garden soil were able to produce high rate of PHA. AK1P and AK2P strains demonstrated high productivity of biopolymer by converting 5% (w/v) lactose as the only carbon source to PHA during fermentation. AY2P strain converted 5% (w/v) of glucose with less PHA accumulation. The favorite temperature for those strains to produce a high rate of PHA was at 30° C.
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