Consolidated bioprocessing (CBP) is a system in which cellulase production, substrate hydrolysis, and fermentation are accomplished in a single process step by cellulolytic microorganisms. CBP offers the potential for lower biofuel production costs due to simpler feedstock processing, lower energy inputs, and higher conversion efficiencies than separate hydrolysis and fermentation processes, and is an economically attractive near-term goal for "third generation" biofuel production. In this review article, production of third generation biofuels from cellulosic feedstocks will be addressed in respect to the metabolism of cellulolytic bacteria and the development of strategies to increase biofuel yields through metabolic engineering.
It is important to determine the effect of changing environmental conditions on the microbial kinetics for design and modeling of biological treatment processes. In this research, the kinetics of nitrate and nitrite reduction by autotrophic hydrogen-dependent denitrifying bacteria and the possible role of acetogens were studied in two sequencing batch reactors (SBR) under varying pH and temperature conditions. A zero order kinetic model was proposed for nitrate and nitrite reduction and kinetic coefficients were obtained at two temperatures (25 +/- 1 and 12 +/- 1 degrees C), and pH ranging from 7 to 9.5. Nitrate and nitrite reduction was inhibited at pH of 7 at both temperatures of 12 +/- 1 and 25 +/- 1 degrees C. The optimum pH conditions for nitrate and nitrite reduction were 9.5 at 25 +/- 1 degrees C and 8.5 at 12 +/- 1 degrees C. Nitrate and nitrite reduction rates were compared, when they were used separately as the sole electron acceptor. It was shown that nitrite reduction rates consistently exceeded nitrate reduction rates, regardless of temperature and pH. The observed transitional accumulation of nitrite, when nitrate was used as an electron acceptor, indicated that nitrite reduction was slowed down by the presence of nitrate. No activity of acetogenic bacteria was observed in the hydrogenotrophic biomass and no residual acetate was detected, verifying that the kinetic parameters obtained were not influenced by heterotrophic denitrification and accurately represented autotrophic activity.
When attempting to increase yields of desirable end-products during fermentation, there is the possibility that increased concentrations of one product redirects metabolism towards the synthesis of less desired products. Changes in growth, final end-product concentrations, and activities of enzymes involved in pyruvate catabolism and fermentative end-product formation were studied in Clostridium thermocellum in response to the addition of individual end-products (H(2), acetate, ethanol, formate, and lactate) to the growth medium. These were added to the growth medium at concentrations ten times greater than those found at the end of growth in cultures grown under carbon-limited conditions using cellobiose (1.1 gl(-1)) as model soluble substrate. Although growth rate and final cell biomass decreased significantly with the addition of all end-products, addition of individual end-products had less pronounced effects on growth. Metabolic shifts, represented by changes in final end-product concentrations, were observed; H(2) and acetate yields increased in the presence of exogenous ethanol and lactate, while ethanol yields increased in the presence of exogenous hydrogen (H(2)), acetate, and lactate. Late exponential phase enzyme activity data of enzymes involved in pyruvate catabolism and end-product formation revealed no changes in enzyme levels greater than 2-fold in response to the presence of any given end-product, with the exception of pyruvate:formate lyase (PFL), ferredoxin-dependent hydrogenase (Fd-H(2)ase), and pyruvate:ferredoxin oxidoreductase (PFO): PFL and Fd-H(2)ase activities increased 2-fold in the presence of ethanol, while PFO activity decreased by 57% in the presence of sodium formate. Changes in enzyme levels did not necessarily correlate with changes in final end-product yields, suggesting that changes in final end-product yields may be governed by thermodynamic considerations rather than levels of enzyme expressed under the conditions tested. We demonstrate that bacterial metabolism may be manipulated in order to selectively improve desired product yields.
Ackerman, J. N., Zvomuya, F., Cicek, N. and Flaten, D. 2013. Evaluation of manure-derived struvite as a phosphorus source for canola. Can. J. Plant Sci. 93: 419–424. There is growing interest in the treatment of swine manure to mitigate water quality issues related to phosphorus (P) from livestock operations. Precipitation of P as struvite (MgNH4PO4·6H2O) is a potential strategy to achieve this. The overall objective of this growth room study was to evaluate the effect of manure-derived struvite (MDS) on canola growth and P recovery efficiency. Pure struvite (PS), monoammonium phosphate (MAP), and polymer-coated monoammonium phosphate (PCMAP) were applied to canola plants in plastic pots containing 2 kg of a sandy loam soil. Biomass yields for MDS and PS were similar at all P rates (mean = 7.6 g pot−1) and significantly smaller than those for MAP and PCMAP (mean = 9.3 g pot−1). Differences in P uptake among P sources were detected at the highest P rate where P uptake was significantly greater for MAP and PCMAP (mean = 22.7 mg P pot−1) than for the struvite forms (mean = 16.4 mg P pot−1). Our results show that although P uptake was similar for the struvites and commercial fertilizers at P2O5 rates of 38 mg pot−1 or lower, biomass yield per unit of P taken up was smaller for the struvites. This may be due to lower initial solubility of the struvites in the alkaline (pH 7.7) soil used in this experiment, which gave an early stage growth advantage to canola fertilized with MAP and PCMAP. These results suggest that it may be necessary to supplement struvite with soluble P fertilizers, such as MAP, if applied on soils such as that tested in this study.
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