Production of polyhydroxyalkanoates by engineered Halomonas bluephagenesis using starch as a carbon source.

A novel α-amylase Amy03713 was screened and cloned from the starch utilization strain Vibrio alginolyticus LHF01. When heterologously expressed in Escherichia coli, Amy03713 exhibited the highest enzyme activity at 45 °C and pH 7, maintained >50 % of the enzyme activity in the range of 25-75 °C and pH 5-9, and sustained >80 % of the enzyme activity in 25 % (w/v) of NaCl solution, thus showing a wide range of adapted temperatures, pH, and salt concentrations. Halomonas bluephagenesis harboring amy03713 gene was able to directly utilize starch. With optimized amylase expression, H. bluephagenesis could produce poly(3-hydroxybutyrate) (PHB), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), and poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P34HB). When cultured for PHB production, recombinant H. bluephagenesis was able to grow up to a cell dry weight of 11.26 g/L, achieving a PHB titer of 6.32 g/L, which is the highest titer that has been reported for PHB production from starch in shake flasks. This study suggests that Amy03713 is an ideal amylase for PHA production using starch as the carbon source in H. bluephagenesis.

[Effect of carbon source and nitrate concentration on denitrifying dephosphorus removal and variation of ORP].

Effect of added carbon source and nitrate concentration on the denitrifying phosphorus removal by SBR process was systematicaly studied, at the same time the variation of oxidation reductiun potential (ORP) was investigated. The results showed the phosphate release rate and the denitrifying and dephosphorus uptake rate in anoxic phase increased with the high carbon source concentration under anaerobic condition (100-300mg/L). However when the carbon source added in anaerobic phase was high to 300mg/L, the residual COD inhibited the succeed denitrifying dephosphorus uptake. High nitrate concentration (5, 15, 40mg/L) in anoxic phase increased the initial denitrifying dephosphorus rate. Once the nitrate depletes, phosphate uptake changed to phosphate release. Moreover, the time of the turning point occurred later with the higher nitrate addition. ORP can be used as a control parameter of phosphorus release, and it can also indicate the denitrificaiton react degree during the anoxic phosphorus removal but can't be used as control parameter of phosphorus uptake.

[Nitrogen removal from the sewage containing seawater via nitrite pathway].

With control of the concentration of free ammonia (FA), shortened nitrification-denitrification was accomplished in SBR to achieve enhanced nitrogen removal from the sewage containing seawater. Several parameters which included salinity, temperature, pH and NH4(+)-N load were studied to evaluate their effects. The results of experiences indicated that shortened nitrification-denitrification can be accomplished in sewage containing seawater with relatively high salinity. With various salinity, the nitrogen removal efficiency had relationship with the NH4(+)-N load, there should be a lower NH4(+)-N load when the salinity was high. The nitrogen removal efficiency reached above 90% when the NH4(+)-N load was not exceed 0.15 kg/(kg.d). Elevation of the temperature availed to higher nitrogen removal efficiency, this efficiency doubled when the reaction temperature was changed from 20 degrees C to 30 degrees C. Relatively high value of pH, in the range of 7.5 and 8.5 had advantage to achieve effective shortened nitrification-denitrification which caused by the selective inhibition of free ammonia(FA).