Produciton Of Thailandepsin A From Burkholderia Thailandensis E264

Rare Burkholderia thailandensis E264 was isolated from the rice field of Thailand. It can produce Thailandepsin A which contains a bicyclic peptide structure. Thailandepins A is a class of histone deacetylase inhibitors (HDACI) and a structural analogue of anti-cancer drugs FK228 with significant role in T-cell lymphoma and skin cancer therapy. Currently,Thailandepsin A yield produced by wild-type strain is very low (10mg / L) and is not suitable for large scale commercial applications. This paper attempts to employ fermentaition medium optimization and macroporous resin HP-20 situ extraction methods to improve Thailandepins A production.In this study,We primarily screened out the proper age of seed, liquid volume (ml), inoculum (%), temperature, pH and rotary speed using one-factor-at-a-time screening. The medium components, including glucose, tryptone, phosphate and sodium citrate had significant effects on Thailandespin A production.It is effective to change Thailandepsin A distribution in the fermentation broth by adding HP-20 resin in broth.The in situ extraction effect of macroporous resins on Thailandespin A decreased the negative feedback inhibition and provided a simpler separation step . The results showed that through addition of maeroporous resin HP-20 , the yield of Thailandepsin A was significantly increased by 80%.It gave a preferable yield when additive amount of resin is 4%(w/v),addtime is 12h and extract liquor is chloroform.Statistics based experimental designs were applied to optimize the culture medium components for enhancing Thailandepsin A production by Burkholderia thailandensis E264. Based on a 24-1 fractional factorial design and a central composite design, an optimum concentration of glucose 17.89 g/L, typtone 34.98 g/L, K2HPO4 20.44 g/L, KH2PO4 4.4g/L and sodium citrate 0.01 g/L was obtained by response surface analysis. The predicted maximum Thailandepsin A yield was as high as 251.9 mg/L, and was further verified by validation experiments. 236.7mg/L Thailandespin A, was almost 2 fold compared to 104mg/L Thailandepsin A from one factor at a time optimization.Thailandespin A fermentation kinetics was also investigated. Kinetic models based on the modified Logistic and Luedeking-Piret equations were developed, providing a good description of temporal variations of biomass(X), product (P) and substrate(S) in Thailandepsin A fermentation.These models were applicable in batch fermentations with initial glucose concentration at about 15g/L.In a laboratory scale stirred tank bioreactor with a working volume of 3 L, the Thailandepsin A fermentation was studied to investigate the effect of macroporous resin in fermentor.In fermentation process, production of Thailandepsin A in the bioreactor added HP-20 resin was increased over 29% than the blank control.