The degradation of (RS)-MCPP was investigated in an anaerobic membrane bioreactor (AnMBR) using nitrate as an available electron acceptor under different COD/NO(3)(-)-N ratios. Results showed high soluble COD removal efficiency (80-93%) when the reactor was operated at high COD/NO(3)(-)-N ratios. However, the COD removal started to decline (average 15%) at high nitrate concentrations coinciding with a drop in nitrate removal efficiency to 37%, suggesting that the denitrification activity dropped and affected the AnMBR performance when nitrate was the predominant electron acceptor. Additionally, the removal efficiency of (RS)-MCPP increased from 2% to 47% with reducing COD/NO(3)(-)-N ratios, whilst the (RS)-MCPP specific utilisation rate (SUR) was inversely proportional to the COD/NO(3)(-)-N ratio, suggesting that a lower COD/NO(3)(-)-N ratios had a positive influence on the (RS)-MCPP SUR. Although nitrate had a major impact on methane production rates, the methane composition was stable (approximately 80%) for COD/NO(3)(-)-N ratios of 23 or more.
An optimized cultivation condition is needed to maximize the functional green fluorescent protein (GFP) production. Six process variables (agitation rate, temperature, initial medium pH, concentration of inducer, time of induction, and inoculum density) were screened using the fractional factorial design. Three variables (agitation rate, temperature, and time of induction) exerted significant effects on functional GFP production in E. coli shake flask cultivation and were optimized subsequently using the Box-Behnken design. An agitation rate of 206 rpm at 31°C and induction of the protein expression when the cell density (OD(600nm)) reaches 1.04 could enhance the yield of functional GFP production from 0.025 g/L to 0.241 g/L, which is about ninefold higher than the unoptimized conditions. Unoptimized cultivation conditions resulted in protein aggregation and hence reduced the quantity of functional GFP. The model and regression equation based on the shake flask cultivation could be applied to a 2-L bioreactor for maximum functional GFP production.