Immobilization is an effective method to increase ethanol production, as proven by previous research. Results almost exclusively demonstrate an increase in ethanol production by and decrease in reproduction rate of immobilized Saccharomyces cerevisiae cells. Recently, research has been conducted to determine the cause of this change. The extreme variance in results due to lack of technology makes it difficult to determine the cellular changes induced by immobilization. With the advent of new technology, specifically gene expression analysis, the RNA content of cells can be easily and rapidly analyzed. S. cerevisiae cells were immobilized in 3% (w/v) calcium alginate beads and grown inside of a packed bed reactor for comparison to planktonic cells growing in batch and chemostat cultures. Temperature inside of the reactor was maintained at 33 C with a pH of 5.5. Cell concentration inside of the beads was monitored periodically in order to create growth curves. Bud scar numbers of immobilized cells were also counted and compared to suspended cells. Scanning electron microscopy images of the alginate beads were taken to determine cell growth inside of the beads. Affymetrix Yeast 2.0 gene chips were used, and the data retrieved was analyzed with GeneSpring software using the Bioconductor packages. Results indicated changes in expression of 3,559 genes with significant difference among treatments by a factor of 2-fold or greater. One-way ANOVA of the filtered data yielded 380 highly significantly different genes between immobilized and suspended cells. Many of the genes pertaining to glycolysis exhibited increased expression levels. Several genes necessary for reproduction were expressed at lower levels in the immobilized cells than in their planktonic counterparts. Many different gene ontologies are discussed, and the expressed genes are mapped onto biochemical pathways.

Hydrogen sulphide (H2S) is a serious problem for many industries, including oil production and processing, pulp and paper, and wastewater treatment. In addition, H2S is usually present in natural gas and biogas. It is necessary to control the generation and release of H2S into the environment because H2S is corrosive, toxic, and has an unpleasant odour. In addition, the removal of H2S from natural gas and biogas is essential for preventing the emission of SO2 upon combustion of these gases. Physicochemical processes have been developed for the removal of H2S. These processes employ techniques such as chemical or physical absorption, thermal and catalytic conversion, and liquid phase oxidation. In comparison, biological processes for the removal of sulphide typically operate at ambient temperature and pressure, with the feasibility for the treatment of smaller streams, and the absence of expensive catalysts. The objective of the present work was to study the biooxidation of sulphide under denitrifying conditions in batch system and a continuous immobilized cell bioreactor using a mixed microbial culture enriched from the produced water of a Canadian oil reservoir. <p>In the batch experiments conducted at various initial sulphide concentrations, an increase in the sulphide oxidation and nitrate reduction rates was observed as the initial sulphide concentration was increased in the range 1.7 to 5.5 mM. An extended lag phase of approximately 10 days was observed when sulphide concentrations around or higher than 14 mM were used. This, when considered with the fact that the microbial culture was not able to oxidize sulphide at an initial concentration of 20 mM, indicates the inhibitory effects of sulphide at high concentrations.<p>The effect of the initial sulphide to nitrate concentrations ratio (ranging from 0.3 to 4.0) was also studied. As the initial sulphide to nitrate ratio decreased, the sulphide oxidation rates increased. The increasing trend was observed for initial nitrate concentrations in the range of 1.3 to 7.3 mM, corresponding to ratios of 4.08 to 0.83. The increase in nitrate reduction rates was more pronounced than that of the sulphide oxidation rates. However at nitrate concentrations higher than 7.3 mM (ratios lower than 0.83) the nitrate reduction rate remained constant. The percentage of sulphide that was oxidized to sulphate increased from 2.4% to 100% as the initial sulphide to nitrate ratio decreased from 4.08 to 0.42. This indicated that at ratios lower than 0.42, nitrate would be in excess and at ratios exceeding 4.08, nitrate would be limiting. In the continuous bioreactor systems, at sulphide loading rates ranging from 0.26 to 30.30 mM/h, sulphide conversion remained in the range of 97.6% to 99.7%. A linear increase in the volumetric oxidation rate of sulphide was observed as the sulphide loading rate was increased with the maximum rate being 30.30 mM/h (98.5% conversion). Application of immobilized cells led to a significant increase in oxidation rate of sulphide when compared with the rates obtained in a bioreactor with freely suspended cells. At nitrate loading rates ranging from 0.19 to 24.44 mM/h, the nitrate conversion ranged from 97.2% to 100% and a linear increase in volumetric reduction rate was observed as the nitrate loading rate was increased, with the maximum rate being 24.44 mM/h (99.7% conversion). <p>A second bioreactor experiment was conducted to investigate the effects of sulphide to nitrate concentrations ratio on the performance of the system. Sulphide conversion was complete at sulphide to nitrate ratios of 1.1 and 1.3, but decreased to 90.5% at the ratio of 3.1 and 65.0% at the ratio of 5.0, indicating nitrate was limiting for sulphide to nitrate ratios of 3.1 and 5.0. The increase in the sulphide to nitrate ratio (and the resulting limitation of nitrate) caused a decrease in the volumetric reaction rate of sulphide.<p>Nitrate conversion was complete at sulphide to nitrate ratios of 1.3, 3.1, and 5.0; however, at a ratio of 1.1, the conversion of nitrate dropped to 59.6%, indicating that nitrate was in excess, and sulphide was limiting. The volumetric reaction rate of nitrate decreased as the sulphide to nitrate ratio increased for ratios of 1.3, 3.1, and 5.0; this was due to the decrease in the nitrate loading rate. For sulphide to nitrate ratios of 1.1 and 1.3, 7.2% and 19.6% of the sulphide was converted to sulphate, respectively. At ratios of 3.1 and 5.0, no sulphate was generated. For ratios between 1.3 and 5.0, an increase in the ratio caused a decrease in the generation of sulphate.

Hydrogen sulphide (H2S) is a serious problem for many industries, including oil production and processing, pulp and paper, and wastewater treatment. In addition, H2S is usually present in natural gas and biogas. It is necessary to control the generation and release of H2S into the environment because H2S is corrosive, toxic, and has an unpleasant odour. In addition, the removal of H2S from natural gas and biogas is essential for preventing the emission of SO2 upon combustion of these gases. Physicochemical processes have been developed for the removal of H2S. These processes employ techniques such as chemical or physical absorption, thermal and catalytic conversion, and liquid phase oxidation. In comparison, biological processes for the removal of sulphide typically operate at ambient temperature and pressure, with the feasibility for the treatment of smaller streams, and the absence of expensive catalysts. The objective of the present work was to study the biooxidation of sulphide under denitrifying conditions in batch system and a continuous immobilized cell bioreactor using a mixed microbial culture enriched from the produced water of a Canadian oil reservoir. <p>In the batch experiments conducted at various initial sulphide concentrations, an increase in the sulphide oxidation and nitrate reduction rates was observed as the initial sulphide concentration was increased in the range 1.7 to 5.5 mM. An extended lag phase of approximately 10 days was observed when sulphide concentrations around or higher than 14 mM were used. This, when considered with the fact that the microbial culture was not able to oxidize sulphide at an initial concentration of 20 mM, indicates the inhibitory effects of sulphide at high concentrations.<p>The effect of the initial sulphide to nitrate concentrations ratio (ranging from 0.3 to 4.0) was also studied. As the initial sulphide to nitrate ratio decreased, the sulphide oxidation rates increased. The increasing trend was observed for initial nitrate concentrations in the range of 1.3 to 7.3 mM, corresponding to ratios of 4.08 to 0.83. The increase in nitrate reduction rates was more pronounced than that of the sulphide oxidation rates. However at nitrate concentrations higher than 7.3 mM (ratios lower than 0.83) the nitrate reduction rate remained constant. The percentage of sulphide that was oxidized to sulphate increased from 2.4% to 100% as the initial sulphide to nitrate ratio decreased from 4.08 to 0.42. This indicated that at ratios lower than 0.42, nitrate would be in excess and at ratios exceeding 4.08, nitrate would be limiting. In the continuous bioreactor systems, at sulphide loading rates ranging from 0.26 to 30.30 mM/h, sulphide conversion remained in the range of 97.6% to 99.7%. A linear increase in the volumetric oxidation rate of sulphide was observed as the sulphide loading rate was increased with the maximum rate being 30.30 mM/h (98.5% conversion). Application of immobilized cells led to a significant increase in oxidation rate of sulphide when compared with the rates obtained in a bioreactor with freely suspended cells. At nitrate loading rates ranging from 0.19 to 24.44 mM/h, the nitrate conversion ranged from 97.2% to 100% and a linear increase in volumetric reduction rate was observed as the nitrate loading rate was increased, with the maximum rate being 24.44 mM/h (99.7% conversion). <p>A second bioreactor experiment was conducted to investigate the effects of sulphide to nitrate concentrations ratio on the performance of the system. Sulphide conversion was complete at sulphide to nitrate ratios of 1.1 and 1.3, but decreased to 90.5% at the ratio of 3.1 and 65.0% at the ratio of 5.0, indicating nitrate was limiting for sulphide to nitrate ratios of 3.1 and 5.0. The increase in the sulphide to nitrate ratio (and the resulting limitation of nitrate) caused a decrease in the volumetric reaction rate of sulphide.<p>Nitrate conversion was complete at sulphide to nitrate ratios of 1.3, 3.1, and 5.0; however, at a ratio of 1.1, the conversion of nitrate dropped to 59.6%, indicating that nitrate was in excess, and sulphide was limiting. The volumetric reaction rate of nitrate decreased as the sulphide to nitrate ratio increased for ratios of 1.3, 3.1, and 5.0; this was due to the decrease in the nitrate loading rate. For sulphide to nitrate ratios of 1.1 and 1.3, 7.2% and 19.6% of the sulphide was converted to sulphate, respectively. At ratios of 3.1 and 5.0, no sulphate was generated. For ratios between 1.3 and 5.0, an increase in the ratio caused a decrease in the generation of sulphate.

Butanol production by four different Clostridium sp. strains was investigated using glucoseP2-medium supplemented with increasing concentrations of butyric acid, added as cosubstrate.Batch fermentations were carried out in serum bottles (freely-suspended cellcultures) and fibrous-bed bioreactor (FBB) with medium recirculation (immobilized cells).Butyric acid clearly revealed to inhibit cellular growth with all specific growth rates decliningupon the increase of butyrate concentrations. However, the presence of low and moderatelevels in the medium can readily enhance the ABE-fermentation and increase butanolproduction through a shift induction towards the solventogenic phase controlled by themedium pH. In all cases it was found that 4.0 g⋅l-1 is the optimal concentration of butyratethat maximizes the yields for all ABE-solvents and butanol productivities. The non-mutant C.acetobutylicum ATCC 824 was singled out as the most efficient butanol productive strainamong all bacteria tested (10.3 g⋅l-1 butanol versus 0.72 g⋅l-1 with and without 4.0 g⋅l-1butyrate, respectively) showing a productivity augment in the order of 0.078 g⋅l-1⋅h-1 (78.5%)and yields of 0.3 g⋅g-1 from substrate and 7.6 g⋅g-1 from biomass versus 0.072 g⋅g-1 and 0.41g⋅g-1 with and without the optimal butyrate concentration, respectively. This strain alsorevealed the best overall tolerance over increasing butyrate concentrations up to ∼6.0 g⋅l-1 andthe highest glucose uptake (65.5%) among all bacteria. Furthermore, the beneficial effects ofbutyric acid were also observed through the use of a fibrous bed-bioreactor when the mutatedstrains of C. beijerinckii ATCC 55025 and BA 101 were tested. The use of this immobilizedcell system effectively improved butanol production over the free system with butanol titersin the fermentation broth around 11.5 g⋅l-1 and 9.4 g⋅l-1 for the two bacteria, respectively,roughly doubling the values attained with the corresponding suspended cell cultures when themedia were supplemented with 4.0 g⋅l-1 of butyrate. All these results confirm theenhancement of butanol formation using either free or immobilized cell culturessupplemented with butyric acid concentrations up to 4.0 g⋅l-1 in the media.