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1.
The inadequate and indiscriminate disposal of sugarcane vinasse in soils and water bodies has received much attention since decades ago, due to environmental problems associated to this practice. Vinasse is the final by-product of the biomass distillation, mainly for the production of ethanol, from sugar crops (beet and sugarcane), starch crops (corn, wheat, rice, and cassava), or cellulosic material (harvesting crop residues, sugarcane bagasse, and wood). Because of the large quantities of vinasse produced, alternative treatments and uses have been developed, such as recycling of vinasse in fermentation, fertirrigation, concentration by evaporation, and yeast and energy production. This review was aimed at examining the available data on the subject as a contribution to update the information on sugarcane vinasse, from its characteristics and chemical composition to alternatives uses in Brazil: fertirrigation, concentration by evaporation, energy production; the effects on soil physical, chemical and biological properties; its influence on seed germination, its use as biostimulant and environmental contaminant. The low pH, electric conductivity, and chemical elements present in sugarcane vinasse may cause changes in the chemical and physical–chemical properties of soils, rivers, and lakes with frequent discharges over a long period of time, and also have adverse effects on agricultural soils and biota in general. Thus, new studies and green methods need to be developed aiming at sugarcane vinasse recycling and disposal.  相似文献   

2.
Fermentation can use renewable raw materials as substrate, which makes it a sustainable method to obtain H2. This study evaluates H2 production by a mixed culture from substrates such as glucose and derivatives from sugarcane processing (sucrose, molasses, and vinasse) combined with landfill leachate. The leachate alone was not a suitable substrate for biohydrogen production. However, leachate blended with glucose, sucrose, molasses, or vinasse increased the H2 production rate by 2.0-, 2.8-, 4.6-, and 0.5-fold, respectively, as compared with the substrates without the leachate. Determination of metals (Cu, Cd, Pb, Hg, Ni, and Fe) at the beginning and at the end of the fermentative assays showed how they were consumed during the fermentation and demonstrated improved H2 production. During fermentation, Cu, Fe, and Cd were the most consumed leachate metals. The best substrate combination to produce H2 was molasses and leachate, which gave high volumetric productivity—469 ml H2/l h. However, addition of the leachate to the substrates stimulated lactic acid formation pathways, which lowered the H2 yield. The use of leachate combined with sugarcane processing derivatives as substrates could add value to the leachate and reduce its polluting power, generating a clean energy source from renewable raw materials.  相似文献   

3.
Fly ash (FA) and vinasse (VN), two industrial wastes, are generated in huge amounts and cause serious hazards to the environment. In this experiment, different proportions of these two wastes were used as food for two epigeic earthworms (Eisenia fetida and Eudrilus eugeniae) to standardize the recycling technique of these two wastes and to study their effect on fungal especially cellulolytic fungal population, cellulase activity and their isozyme pattern, chitin content and microbial biomass of waste mixture during vermicomposting. Increasing VN proportion from 25% to 50% or even higher, counts of both fungi and cellulolytic fungi in waste mixtures were significantly (P ? 0.05) increased during vermicomposting. Higher cellulase activity in treatments having 50% or more vinasse might be attributed to the significantly (P ? 0.05) higher concentration of group I isozyme while concentrations of other isozymes (group II and III) of cellulase were statistically at par. Higher chitin content in vinasse-enriched treatments suggested that fungal biomass and fungi-to-microbial biomass ratio in these treatments were also increased due to vermicomposting. Results revealed that Eudrilus eugeniae and Eisenia fetida had comparable effect on FA and VN mixture during vermicomposting. Periodical analysis of above-mentioned biochemical and microbial properties and nutrient content of final vermicompost samples indicated that equal proportion (1:1, w/w) of FA and VN is probably the optimum composition to obtain best quality vermicompost.  相似文献   

4.
Buckwheat tea waste (BWTW) and barley tea waste (BTW), by-products of the beverage industry, are alternative carbohydrate sources for ethanol production. In this study, optimal enzyme loading for enzymatic saccharification of BWTW and BTW was determined, and simultaneous saccharification and fermentation (SSF) was performed by Saccharomyces cerevisiae and Mucor indicus to produce ethanol. Optimal enzyme loading for enzymatic saccharification of 2?% w/v BWTW and BTW was 0.5?% (weight of enzyme/weight of tea wastes) for BWTW and 0.1?% for BTW. Ethanol production from BWTW by S. cerevisiae and M. indicus after 48?h of SSF was 49.9/100?g of BWTW and 47.9/100?g of BWTW, respectively, with 0.5?% enzyme loading. Ethanol production from BTW by S. cerevisiae and M. indicus after 48?h of SSF was 20.5/100?g of BTW and 21.6/100?g of BTW, corresponding to 62 and 66?% of the theoretical yield based on starch content, respectively, with 0.1?% of enzyme loading. Furthermore, S. cerevisiae produced 76?% of the theoretical yield based on the total glucose from starch in BWTW and BTW when a mixture of BWTW and BTW was used as a substrate, with 0.2?% enzyme loading and no additional nitrogen or mineral sources.  相似文献   

5.
Considerable concern is caused by the problem of potato peel waste (PPW) to potato industries in Europe. An integrated, environmentally-friendly solution is yet to be found and is currently undergoing investigation. Potato peel is a zero value waste produced by potato processing plants. However, bio-ethanol produced from potato wastes has a large potential market. If Federal Government regulations are adopted in light of the Kyoto agreement, the mandatory blending of bio-ethanol with traditional gasoline in amounts up to 10% will result in a demand for large quantities of bio-ethanol. PPW contain sufficient quantities of starch, cellulose, hemicellulose and fermentable sugars to warrant use as an ethanol feedstock. In the present study, a number of batches of PPW were hydrolyzed with various enzymes and/or acid, and fermented by Saccharomyces cerevisae var. bayanus to determine fermentability and ethanol production. Enzymatic hydrolysis with a combination of three enzymes, released 18.5 g L?1 reducing sugar and produced 7.6 g L?1 of ethanol after fermentation. The results demonstrate that PPW, a by-product of the potato industry features a high potential for ethanol production.  相似文献   

6.
The production of hydrogen through Anaerobic Digestion (AD) has been investigated to verify the efficacy of several pretreatment processes. Three types of waste with different carbon structures have been tested to obtain an extensive representation of the behavior of the materials present in Organic Waste (OW). The following types of waste were selected: Sweet Product Residue (SPR), i.e., confectionary residue removed from the market after the expiration date, Organic Waste Market (OWM) refuse from a local fruit and vegetable market, and Coffee Seed Skin (CSS) waste from a coffee production plant. Several pretreatment processes have been applied, including physical, chemical, thermal, and ultrasonic processes and a combination of these processes. Two methods have been used for the SPR to remove the packaging, manual (SPR) and mechanical (SPRex). A pilot plant that is able to extrude the refuse to 200 atm was utilized. Two parameters have been used to score the different pretreatment processes: efficiency (ξ), which takes into account the amount of energy produced in the form of hydrogen compared with the available energy embedded in the refuse, and efficacy (η), which compares the efficiency obtained using the pretreated refuse with that obtained using the untreated refuse. The best result obtained for the SPR was the basic pretreatment, with η = 6.4, whereas the thermal basic pretreatment gave the highest value, η = 17.0 for SPRex. The best result for the OWM was obtained through a combination of basic/thermal pretreatments with η = 9.9; lastly, the CSS residue with ultrasonic pretreatment produced the highest quantity of hydrogen, η = 5.2.  相似文献   

7.
Molasses-based distilleries are one of the most polluting industries generating large volumes of high strength wastewater called vinasse. Different processes covering anaerobic, aerobic as well as physicochemical methods have been employed to treat this effluent. This study evaluated the microbial communities present in the vinasse during different stages of its treatment by traditional and molecular methods. The analysis of the efficiency of each treatment was performed by physicochemical parameters and toxicity analysis. The treatment of vinasse was performed in the following steps: high flow fermentation; filtration; chemical flakes; low-flow fermentation; filtration; and neutralization. The physicochemical analysis in different stages of the vinasse treatment demonstrated that phases of treatment influenced the performance of the evaluated parameters. Among the 37 parameters, 9 were within the limits established by the Commission for Environmental Policy of Minas Gerais, Brazil (COPAM), especially BOD (96.7% of pollution reduction), suspended solids (99.9%), pH, copper (88%), iron (92.9%), and manganese (88%). Some parameters, even after treatment, did not fit the maximum allowed by legislation. The microbial population decreased reaching 3 log CFU/ml present in the steps of the flakes chemical and disinfection treatment of vinasse. Lactobacillus brevis and Pichia kudriavzevii were present in all stages of the treatments, showing that these microorganisms were resistant and demonstrated that they might be important in the treatment of vinasse. The vinasse showed a significant reduction of pollution load after the disinfection treatment however still should not be discarded into water bodies because the high values of tannins and sediment solids, but suggest the use of the effluent in the cooling coil during the distillation process of the beverage.  相似文献   

8.
The utilization of bioethanol is being focused on as a fuel alternative to oil and or natural gas. Bioethanol production from cellulosic plant residues is one of the solutions proposed for the problems caused by usage of food crops that are also vital for human consumption, such as sugar cane and corn, as a source of bioethanol. However, to utilize these new sources for bioethanol production, conditions for saccharification in each different material have not been optimized. In this study, we reported some optimum conditions for the saccharification of Korean lawn grass (KL) and bent grass (BG) using acremonium cellulase and endoglucanase as saccharifying enzymes for ethanol fermentation. With respect to saccharification of KL and BG, 0.19 and 0.18 g of d-glucose per g-substrate at maximum were produced, respectively. Comminution with a ball mill was found to be effective in the saccharification of KL, while ball-milled BG showed no significant improvement in saccharification. Being incorporated with 99 % of d-glucose consumption, saccharified KL was incubated for 3 days with Saccharomyces cerevisiae and Zymomonas mobilis, respectively, and each mixture fermented to ethanol yielding approximately 100 % of theoretical values from d-glucose consumption, respectively.  相似文献   

9.
Present study describes the treatment of molasses spentwash and its use as a potential low cost substrate for production of biopolymer polyhydroxybutyrate (PHB) by waste activated sludge. Fluorescence microscopy revealed the presence of PHB granules in sludge biomass which was further confirmed by fourier transform-infra-red spectroscopy (FT-IR) and 13C nuclear magnetic resonance (NMR). The processing of molasses spentwash was carried out for attaining different ratios of carbon and nitrogen (C:N). Highest chemical oxygen demand (COD) removal and PHB accumulation of 60% and 31% respectively was achieved with raw molasses spentwash containing inorganic nitrogen (C:N ratio = 28) followed by COD removal of 52% and PHB accumulation of 28% for filtered molasses containing inorganic nitrogen (C:N ratio = 29). PHB production yield (Yp/s) was highest (0.184 g g?1 COD consumed) for deproteinized spentwash supplemented with nitrogen. In contrast, the substrate consumption and product formation were higher in case of raw spentwash. Though COD removal was lowest from deproteinized spentwash, evaluation of kinetic parameters suggested higher rates of conversion of available carbon to biomass and PHB. Thus the process provided dual benefit of conversion of two wastes viz. waste activated sludge and molasses spentwash into value-added product-PHB.  相似文献   

10.
A pot experiment is described with a fast-growing poplar clone and two native willows (Populus deltoides Bartr. cl. I-69/55 (Lux)), Salix viminalis L. and Salix purpurea L.), irrigated with landfill leachate and compost wastewater over a 1-year growing period. The use of leachate resulted in up to 155% increased aboveground biomass compared to control water treatments and in up to 28% reduced aboveground biomass compared to a complete nutrient solution. The use of compost wastewater resulted in up to 62% reduced aboveground biomass compared to the control treatments and in up to 86% reduced aboveground biomass compared to the complete nutrient solution. Populus was the most effective in biomass production due to the highest leaf production, whereas S. purpurea was the least effective in biomass accumulation, but less sensitive to high ionic strength of the irrigation water compared to S. viminalis. The results showed a high potential for landfill leachate application (with up to 2144 kg N ha?1, 144 kg P ha?1, 709 kg K ha?1, 1010 kg Cl ha?1, and 1678 kg Na ha?1 average mass load in the experiment). High-strength compost wastewater demonstrated less potential for application as irrigation and fertilization source even in high water-diluted treatments (1:8 by volume).  相似文献   

11.
This study investigated the production of d-lactic acid from unutilized sugarcane bagasse using steam explosion pretreatment. The optimal steam pressure for a steaming time of 5?min was determined. The steam-exploded sugarcane bagasse was hydrolyzed using cellulase (Meicelase) and then the hydrolyzate was subjected to fermentation substrate. By enzymatic saccharification using Meicelase, the highest recovery of glucose from raw bagasse, 73.7?%, was obtained at a steam pressure of 20?atm. For extracted residue with water after steam explosion, the glucose recovery increased up to 94.9?% at a steam pressure of 20?atm. These results showed that washing with water is effective in removing enzymatic reaction inhibitors. After steam pretreatment (steam pressure of 20?atm), d-lactic acid was produced by Lactobacillus delbrueckii NBRC 3534 from the enzymatic hydrolyzate of steam-exploded bagasse and washed residue. The conversion rate of d-lactic acid obtained from the glucose concentration was 66.6?% for the hydrolyzate of steam-exploded bagasse without washing with water and 90.0?% for that derived from the extracted residue with water after steam explosion. These results also demonstrated that the hydrolyzate of steam-exploded bagasse (without washing with water) contains fermentation inhibitors and washing with water can remove them.  相似文献   

12.
Dried and ground banana peel biomass (BP) after hydrothermal sterilization pretreatment was used for ethanol production using simultaneous saccharification and fermentation (SSF). Central composite design (CCD) was used to optimize concentrations of cellulase and pectinase, temperature and time for ethanol production from BP using SSF. Analysis of variance showed a high coefficient of determination (R2) value of 0.92 for ethanol production. On the basis of model graphs and numerical optimization, the validation was done in a laboratory batch fermenter with cellulase, pectinase, temperature and time of nine cellulase filter paper unit/gram cellulose (FPU/g-cellulose), 72 international units/gram pectin (IU/g-pectin), 37 °C and 15 h, respectively. The experiment using optimized parameters in batch fermenter not only resulted in higher ethanol concentration than the one predicted by the model equation, but also saved fermentation time. This study demonstrated that both hydrothermal pretreatment and SSF could be successfully carried out in a single vessel, and use of optimized process parameters helped achieve significant ethanol productivity, indicating commercial potential for the process. To the best of our knowledge, ethanol concentration and ethanol productivity of 28.2 g/l and 2.3 g/l/h, respectively from banana peels have not been reported to date.  相似文献   

13.
The filamentous alga Hydrodictyon reticulatum harvested from a bench-scale wastewater treatment pond was used to evaluate biogas production after ultrasound pretreatment. The effects of ultrasound pretreatment at a range of 10–5000 J/mL were tested with harvested H. reticulatum. Cell disruption by ultrasound was successful and showed a higher degree of disintegration at a higher applied energy. The range of 10–5000 J/mL ultrasound was able to disintegrated H. reticulatum and the soluble COD was increased from 250 mg/L to 1000 mg/L at 2500 J/mL. The disintegrated algal biomass was digested for biogas production in batch experiments. Both cumulative gas generation and volatile solids reduction data were obtained during the digestion. Cell disintegration due to ultrasound pretreatment increased the specific biogas production and degradation rates. Using the ultrasound approach, the specific methane production at a dose of 40 J/mL increased up to 384 mL/g-VS fed that was 2.3 times higher than the untreated sample. For disintegrated samples, the volatile solids reduction was greater with increased energy input, and the degradation increased slightly to 67% at a dose of 50 J/mL. The results also indicate that disintegration of the algal cells is the essential step for efficient anaerobic digestion of algal biomass.  相似文献   

14.
Bacterial cellulose pellicles were produced by Gluconacetobacter xylinus using non conventional low-cost carbon sources, such as glycerol remaining from biodiesel production and grape bagasse, a residue of wine production. The carbon sources assayed showed their suitability for microbial cellulose production, with relatively high production values such as 10.0 g/l for the culture medium with glycerol from biodiesel as carbon source and corn steep liquor as nitrogen source; and 8.0 g/l for the culture medium containing grape bagasse and corn steep liquor. Glucose, commercial glycerol and cane molasses were also assayed as carbon sources for comparison. The bacterial celluloses produced were characterized by means of scanning electron microscopy, X-ray diffraction, Fourier transform infrared spectroscopy and thermogravimetric analysis. Morphological analysis showed that bacterial cellulose microfibrils produced from the non-conventional media used were several micrometers long and had rectangular cross-sections with widths and thicknesses in the range of 35–70 and 13–24 nm, respectively. X-ray patterns showed crystallinity levels in the range of 74–79 % (area method), whereas both X-ray patterns and infrared spectroscopy evidenced the presence of peaks characteristic of Cellulose I polymorph. Besides thermal properties were similar to those found for the pellicle obtained from glucose. The study performed showed the suitability of using wine residues or glycerol remaining from increasing biodiesel production as cheap carbon sources for production of bacterial cellulose microfibrils, with similar characteristics as those obtained by use of more expensive carbon sources such as glucose or commercial glycerol. On the other hand, the low cost nitrogen sources used (corn steep liquor or diammonium phosphate) also contributed to the economy of the bioprocess.  相似文献   

15.
Mineralization potentials are often used to classify organic wastes. These methods involve measuring CO2 production during batch experiments, so variations in chemical compounds are not addressed. Moreover, the physicochemical conditions are not monitored during the reactions. The present study was designed to address these deficiencies. Incubations of a mixture of soil and waste (vinasse at 20% dry matter from a fermentation industry) were conducted in aerobic and anaerobic conditions, and liquid samples obtained by centrifugation were collected at 2 h, 1 d and 28 d. Dissolved organic carbon (DOC) patterns highlighted that: there was a “soil effect” which increased organic matter (OM) degradation in all conditions compared to vinasse incubated alone; and OM degradation was faster under aerobic conditions since 500 mg kg?1 of C remained after aerobic incubation, as compared to 4000 mg kg?1 at the end of the anaerobic incubation period. No changes were detected by Fourier transform infrared spectroscopy (FTIR) between 2 h and 1 d incubation. At 28 days incubation, the FTIR signal of the aerobic samples was deeply modified, thus confirming the high OM degradation. Under anaerobic conditions, the main polysaccharide contributions (ν(C–O)) disappeared at 1000 and 1200 cm?1, as also confirmed by the 13C NMR findings. Under aerobic incubation, a 50% decrease in the polysaccharide proportion was observed. Under anaerobic conditions, significant chemical modifications of the organic fraction were detected, namely formation of low molecular weight organic acids.  相似文献   

16.
Biodiesel can be a potential alternative to petroleum diesel, but its high production cost has impeded its commercialization in most parts of the world. One of the main drivers for the generation and use of biodiesel is energy security, because this fuel can be produced from locally available resources, thereby reducing the dependence on imported oil. Many countries are now trying to produce biodiesel from plant or vegetable oils. However, the consumption of large amounts of vegetable oils for biodiesel production could result in a shortage in edible oils and cause food prices to soar. Alternatively, the use of animal fat, used frying oils, and waste oils from restaurants as feedstock could be a good strategy to reduce the cost. However, these limited resources might not meet the increasing demand for clean, renewable fuels. Therefore, recent research has been focused the use of residual materials as renewable feedstock in order to lower the cost of producing biodiesel. Microbial oils or single cell oils (SCOs), produced by oleaginous microorganisms have been studied as promising alternatives to vegetable or seed oils. Various types of agro-industrial residues have been suggested as prospective nutritional sources for microbial cultures. Since the most abundant residue from agricultural crops is lignocellulosic biomass (LCB), this byproduct has been given top-priority consideration as a source of biomass for producing biodiesel. But the biological transformation of lignocellulosic materials is complicated due to their crystalline structure. So, pretreatment is required before they can be converted into fermentable sugar. This article compares and scrutinizes the extent to which various microbes can accumulate high levels of lipids as functions of the starting materials and the fermentation conditions. Also, the obstacles associated with the use of LCB are described, along with a potentially viable approach for overcoming the obstacles that currently preclude the commercial production of biodiesel from agricultural biomass.  相似文献   

17.
The present study examined the suitability of vermicomposting technology for processing source-separated human faeces. Since the earthworm species Eisenia fetida could not survive in fresh faeces, modification in the physical characteristics of faeces was necessary before earthworms could be introduced to faeces. A preliminary study with six different combinations of faeces, soil and bulking material (vermicompost) in different layers was conducted to find out the best condition for biomass growth and reproduction of earthworms. The results indicated that SVFV combination (soil, vermicompost, faeces and vermicompost – bottom to top layers) was the best for earthworm biomass growth indicating the positive role of soil layer in earthworm biomass growth. Further studies with SVFV and VFV combinations, however, showed that soil layer did not enhance vermicompost production rate. Year-long study conducted with VFV combination to assess the quality and quantity of vermicompost produced showed an average vermicompost production rate of 0.30 kg-cast/kg-worm/day. The vermicompost produced was mature as indicated by low dissolved organic carbon (2.4 ± 0.43 mg/g) and low oxygen uptake rate (0.15 ± 0.09 mg O2/g VS/h). Complete inactivation of total coliforms was noted during the study, which is one of the important objectives of human faeces processing. Results of the study thus indicated the potential of vermicomposting for processing of source-separated human faeces.  相似文献   

18.
An attempted has been made to recover high-calorific fuel gas and useful carbonaceous residue by the electric arc pyrolysis of waste lubricating oil. The characteristics of gas and residues produced from electric arc pyrolysis of waste lubricating oil were investigated in this study. The produced gas was mainly composed of hydrogen (35–40%), acetylene (13–20%), ethylene (3–4%) and other hydrocarbons, whereas the concentration of CO was very low. Calorific values of gas ranged from 11,000 to 13,000 kcal kg?1 and the concentrations of toxic gases, such as NOx, HCl and HF, were below the regulatory emissions limit. Gas chromatography–mass spectrometry (GC/MS) analysis of liquid-phase residues showed that high molecular-weight hydrocarbons in waste lubricating oil were pyrolyzed into low molecular-weight hydrocarbons and hydrogen. Dehydrogenation was found to be the main pyrolysis mechanism due to the high reaction temperature induced by electric arc. The average particle size of soot as carbonaceous residue was about 10 μm. The carbon content and heavy metals in soot were above 60% and below 0.01 ppm, respectively. The utilization of soot as industrial material resources such as carbon black seems to be feasible after refining and grinding.  相似文献   

19.
In this research, a biosurfactant-producing bacterium with capability of asphaltene degradation was isolated from oil-contaminated soil samples, and identified as Bacillus cereus. This strain produced an effective biosurfactant in the presence of molasses and the surface tension was reduced to the level of 36.4 mN/m after 48 h under optimum conditions. The optimum values of carbon-to-nitrogen ratio (C:N), pH, and temperature for biosurfactant production were determined as 30:1, 7.3 and 29 °C, respectively, using response surface methodology. The maximum emulsification activity in the culture broth was 53 % after 48 h using kerosene at 25 °C. The goodness of fit of four growth kinetic models including Tessier, Contois, Logistic and Westerhoff was compared for the bacterial growth and molasses utilization of B. cereus in 5-L batch bioreactor during 120 h. Conducted kinetic study showed that biosurfactant production had a good fit with the Contois growth kinetic model (R2 = 0.962) and the maximum specific growth rate (µ max ), saturation constant (K s ) and the yield of biomass per substrate (Y x/s ) were determined to be 0.145 h?1, 1.83 g/L and 0.428 g/g, respectively. The asphaltene biodegradation in flask was evaluated by FTIR analysis and quantified by a spectrophotometer. This bacterium was able to degrade up to 40 % of asphaltene as a sole carbon and energy source after 60 days at 28 °C. The resulting surface tension of 30.2 mN/m with the critical micelle concentration of 23.4 mg/L indicated good efficiency of the biosurfactant.  相似文献   

20.
The feasibility of adding crude glycerol from the biodiesel industry to the anaerobic digesters treating sewage sludge in wastewater treatment plants was studied in both batch and continuous experiments at 35 °C. Glycerol addition can boost biogas yields, if it does not exceed a limiting 1% (v/v) concentration in the feed. Any further increase of glycerol causes a high imbalance in the anaerobic digestion process. The reactor treating the sewage sludge produced 1106 ± 36 ml CH4/d before the addition of glycerol and 2353 ± 94 ml CH4/d after the addition of glycerol (1% v/v in the feed). The extra glycerol-COD added to the feed did not have a negative effect on reactor performance, but seemed to increase the active biomass (volatile solids) concentration in the system. Batch kinetic experiments showed that the maximum specific utilization rate (μmax) and the saturation constant (KS) of glycerol were 0.149 ± 0.015 h?1 and 0.276 ± 0.095 g/l, respectively. Comparing the estimated values with the kinetics constants for propionate reported in the literature, it can be concluded that glycerol uptake is not the rate-limiting step during the process.  相似文献   

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