Eugenol Case Study

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3.1. Materials and microorganisms Eugenol (99 %); Coniferyl Aldehyde (98 %); Ferulic Acid (99 %); Vanillin (99 %); Vanillic Acid (99 %) were obtained from Sigma Aldrich. Methanol used for high-performance liquid chromatography (HPLC) analysis was purchased from Merck and was HPLC grade. All the other chemicals used were of analytical grade and commercially available. The Pseudomonas fluorescens NCIM 2100 bacterial strain was obtained from National Collection of Industrial Microorganisms (NCIM), Pune and was provided by Department of Bio-Engineering, BIT Mesra. Methodology for screening After subculturing and maintenance of Pseudomonas fluorescens NCIM 2100, obtained from departmental culture stock, Starter bacterial culture of known microorganisms…show more content…
Medium and culture conditions Cells were grown in a 100-mL flask containing 25 mL into minimal medium supplemented with Eugenol for 2, 4 and 6 days incubation at 37 °C. The following Table 3. represents the composition of the modified minimal media used for biotransformation of eugenol (Muheim and Lerch, 1999). The pH of the medium was adjusted between 7.0 to 7.25 and autoclaved to obtain sterilized media for further…show more content…
Effect of Various nitrogen sources: The modified minimal media used for biotransformation of Eugenol has yeast extract (0.5gm/L) as its sole nitrogen source. This was replaced by beef extract, peptone, ammonium nitrate and sodium nitrate respectively at 0.5gm/L to monitor effect of various organic and inorganic nitrogen sources. Culture was kept under optimum temperature and pH for an incubation period of 2, 4, and 6 days. 3.5.5. Effect of varying temperature and pH: Effect of varying temperature (5 °C, 25 °C, 37 °C and 45 °C) on biotransformation of Eugenol was studied at pH of 7. After choosing an optimum temperature, pH was optimized in the range of 5-9 to observe its effect on production of valuable products. Similarly, Medium and culture conditions were maintained as described in section 4.2. The cultures were incubated for 2, 4, and 6 days to monitor effect of varying temperature and pH on production of different metabolites during biotransformation. 3.5.6. Effect of inoculum size (v/v %): Effect of different inoculum size (2 %, 3 %, 4 %, 5 % and 6 %) on production of desired metabolites by the selected microorganism was studied. The modified minimal media used for biotransformation of optimized Eugenol and other different parameters was studied for the effect of different inoculum size (v/v %). Culture was kept under optimum temperature and pH for incubation period of 2, 4 and 6

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