Isolation and Identification of Polyhydroxyalkanoates from two Strains of Clostridium Bifermentans Isolated from the Soil Near the Gas Station in Basrah City
Journal Title: Biomedical Journal of Scientific & Technical Research (BJSTR) - Year 2019, Vol 13, Issue 2
Abstract
Two strains of Clostridium isolated from soilnear the gas stations in Basrah, Iraq, was identified by 16S rRNA sequencing as Clostridium bifermentans strain 0910-0608 and - Clostridium bifermentans strain 0912-02001 Polyhydroxyalkanoates (PHA) production by these strains were investigated. The extracted PHA was characterized by FTIR spectroscopy. It was found that these strains are PHA producers. In recent years, there has been a shift in public opinion with people becoming more ecologically aware. The shift in public opinion has driven industries to investigate biodegradable alternatives to plastic which are not manufactured using petrochemical methods [1,2]. Poly hydroxyl alkanoates (PHAs) are among the top group of biopolymers that have been intensively investigated and commercialized. PHAs produced by a broad range of microorganism. PHA can be synthesized by over 30 % of soil inhabiting bacteria [3]. Accumulating PHAs is a natural way for bacteria to store carbon and energy, when nutrient supplies are imbalanced. These polyesters are accumulated when bacterial growth is limited by depletion of nitrogen, phosphorous [4] or oxygen and an excess amount of a carbon source is still present. As PHAs are insoluble in water, the polymers are accumulated in intracellular granules inside the cells. It is advantageous for bacteria to store excess nutrients inside their cells, especially as their general physiological fitness is not affected. By polymerizing soluble intermediates into insoluble molecules, the cell does not undergo alterations of its osmotic state. Thus, leakage of these valuable compounds out of the cell is prevented and the nutrient stores will remain securely available at a low maintenance cost [5].Samples were collected from the area near the gas stations in Basrah city. Soil samples 5cm deep from surface was used for isolation of the bacteria. About 15-20g of soil samples scraped with sterile plastic jars, and for the isolation of organisms one gram of soil sample is dispensed in 10ml of sterile distilled water. This is mixed vigorously and 0.1ml were pre-cultured on modified PY medium according to [6] the pre- cultivation step was conducted under anaerobic condition using Anaerobic Jar at 30oC for 7days. After incubation period the isolates were studied for morphological and biochemical characteristics. Initially, spread onto blood agare and egg yolk agar plates . The plates were incubated for 24h under anaerobic condition using Anaerobic Jar at the temperature of 30oC. then Gram staining was performed after which biochemical characterization was done to identify the phenotypic characters of the bacterial isolates, indol production , voges- proskauer and citrate utilization.
Authors and Affiliations
Nawres N Jaber
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