Production Of Flavonoid, An Anti-Inflammatory Agent From Trametes Lactinea
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Date
2014-09
Authors
Yahaya, Yus Azila
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Abstract
Flavonoid is one of the remarkable bioactive compound that possesses broad pharmacological activities including anti-inflammatory. A simple and effective bioroute approach for the production of flavonoid by six locally isolated strain obtained from culture collection center were looked at. Trametes lactinea was found to produce the highest flavonoid in its culture broth and inhibited the anti-inflammatory enzyme activities, hyaluronidase and lipoxygenase. In shake flask culture, the effect of culture conditions on biomass and flavonoid production by T. lactinea (media composition, initial pH, inoculum volume, temperature and incubation time) were optimized using one-factor-at-a-time (OFAT) method. Results showed that the highest biomass was obtained in the Medium 3, containing glucose, yeast and peptone. However, this production medium was not favourable for flavonoid production by the tested fungus but it preferred to condition of less carbon media with a small amount of minerals (Medium 2). Four selected parameters obtained in OFAT was further optimized for flavonoid production by T. lactinea using a Response Surface Methodology (RSM) coupled with Box Behnken design (BBD). Four parameters that involved in this study were initial pH, incubation temperature and incubation time. The highest flavonoid production (20.02 μg/mL) was obtained in the production media with initial pH 6, incubation temperature 35oC and incubation time 8.1 days, respectively. To achieve higher biomass and flavonoid production, the performance of 2.5 L stirred tank bioreactor was investigated by considering effect of aeration rates (0.5 to 1.5 vvm), agitation speed (100 to 300 rpm)
and initial glucose concentration 10 to 60 g/L). The highest biomass and flavonoid production were obtained at agitation speed 300 and 100 rpm, and initial glucose concentration at 40 and 20 g/L, respectively. The volumetric oxygen transfer coefficient (kLa) of T. lactinea was also studied by considering the effect of aeration rate and agitation speed. Results showed that kLa values increased with increased of aeration rate from 0.5 to 1.5 vvm and agitation speed from 100 to 300 rpm, respectively. Fed batch fermentations of T. lactinea in a stirred tank bioreactor was also conducted and the results showed that a glucose feeding strategy was favourable process that enhanced the flavonoid production compared to batch mode and a shake flask culture. Kinetics of growth, product formation, glucose utilization and inhibition of the tested fungus were also looked at. For T. lactinea growth, the Logistic, Modified Logistic and Gompertz models fitted well with the experimental data with R2 > 0.98 and RMSD <0.050 at initial glucose concentration at 40 g/L. The modified Gompertz model provided a more accurate description with R2 values > 0.90 and smaller RMSD for the flavonoid production at initial glucose concentration at 20 g/L. For substrate utilization and inhibition, the modified Gompertz model and the Luong model described well the experimental data with high R2 and low RMSD values. Rutin was found in the aqueous extract of T. lactinea when analyzed using various analytical tools (TLC, UV-vis spectrophotometer and HPLC). Cytotoxicity test revealed that the aqueous extract of T. lactinea possessed no toxic effect towards Chinese hamster lung (V79-4) cell lines within 24 hour exposure.
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Flavonoid is one of the remarkable bioactive compound that possesses , broad pharmacological activities including anti-inflammatory.