Glutamate production from CO2 by Marine CyanobacteriumSynechococcus sp.

A photobioreactor was constructed in the form of a Perspex column 900 mm tall with an internal diameter of 70 mm. The reactor volume was 1.8 L and the light source consisted of a metal-halide lamp to reproduce sunlight. Light was distributed through the culture using a new type of optical fiber that diffuses light out through its surface, perpendicular to the fiber axis. A cluster of 661 light-diffusing optical fibers (LDOFs) pass from the light source through the reactor column (60-cm culture depth) and are connected to a mirror at the top of the reactor. This biosolar reactor has been used for the production of glutamate from CO₂ by the marine cyanobacterium Synechococcus sp. NKBG040607. We present here details of the construction of the biosolar reactor and characterization of its properties. The effect of light intensity on glutamate production was measured. Carbon dioxide-to-glutamate conversion ratios were determined at different cell densities: the maximum conversion ratio (28%) was achieved at a cell density of 3x10⁸ cells/mL. A comparison of glutamate production using the LDOF biosolar reactor described here with production by batch culture using free or immobilized cells showed that use of an optical-fiber biosolar reactor increased glutamate-production efficiency 6.75-fold. We conclude that as a result of its high surface-to-volume ratio (692/m) increased photoproduction of useful compounds may be achieved. Such a system is generally applicable to all aspects of photobiotechnology.     

Effect of CO2 Concentration on Growth and Biochemical Composition of Newly Isolated Indigenous Microalga Scenedesmus bajacalifornicus BBKLP-07

Photosynthetic mitigation of CO₂ through microalgae is gaining great importance due to its higher photosynthetic ability compared to plants, and the biomass can be commercially exploited for various applications. CO₂ fixation capability of the newly isolated freshwater microalgae Scenedesmus bajacalifornicus BBKLP-07 was investigated using a 1-l photobioreactor. The cultivation was carried at varying concentration of CO₂ ranging from 5 to 25%, and the temperature and light intensities were kept constant. A maximum CO₂ fixation rate was observed at 15% CO₂ concentration. Characteristic growth parameters such as biomass productivity, specific growth rate, and maximum biomass yield, and biochemical parameters such as carbohydrate, protein, lipid, chlorophyll, and carotenoid were determined and discussed. It was observed that the effect of CO₂ concentration on growth and biochemical composition was quite significant. The maximum biomass productivity was 0.061 ± 0.0007 g/l/day, and the rate of CO₂ fixation was 0.12 ± 0.002 g/l/day at 15% CO₂ concentration. The carbohydrate and lipid content were maximum at 25% CO₂ with 26.19 and 25.81% dry cell weight whereas protein, chlorophyll, and carotenoid contents were 32.89% dry cell weight, 25.07 μg/ml and 6.15 μg/ml respectively at 15% CO₂ concentration.

     

Antioxidant Activity and Kinetic Characterization of Chlorella vulgaris Growth under Flask-Level Photoheterotrophic Growth Conditions

C. vulgaris is a unicellular microalgae, whose growth depends on the conditions in which it is found, synthesizing primary and secondary metabolites in different proportions. Therefore, we analyzed and established conditions in which it was possible to increase the yields of metabolites obtained at the flask level, which could then be scaled to the photobioreactor level. As a methodology, a screening design was applied, which evaluated three factors: type of substrate (sodium acetate or glycerol); substrate concentration; and exposure-time to red light (photoperiod: 16:8 and 8:16 light/darkness). The response variables were: cell division; biomass; substrate consumption; and antioxidant activity in intracellular metabolites (ABTS•+ and DPPH•). As a result, the sodium acetate condition of 0.001 g/L, in a photoperiod of 16 h of light, presented a doubling time (Td = 4.84 h) and a higher rate of division (σ = 0.20 h⁻¹), having a final biomass concentration of 2.075 g/L. In addition, a higher concentration of metabolites with antioxidant activity was found in the sodium acetate (0.629 Trolox equivalents mg/L ABTS•+ and 0.630 Trolox equivalents mg/L DPPH•). For the glycerol, after the same photoperiod (16 h of light and 8 h of darkness), the doubling time (Td) was 4.63 h, with a maximum division rate of σ = 0.18 h⁻¹ and with a biomass concentration at the end of the kinetics of 1.4 g/L. Sodium acetate under long photoperiods, therefore, is ideal for the growth of C. vulgaris, which can then be scaled to the photobioreactor level.     

Spiral tubular bioreactors for hydrogen production by photosynthetic microorganisms

Spiral tubular bioreactors were constructed out of transparent PVC tubing for H2 production applications. Both a cyanobacterialAnabaena variabilis mutant that lacks uptake hydrogenase activity and the photosynthetic bacteriumRhodobacter sp. CBS were tested in the bioreactors. Continuous H₂ photoproduction at an average rate of 19 mL min^-2.h^-1 was observed using theA. variabilis mutant under an air atmosphere (without argon sparging or application of a partial vacuum). The cyanobacterial photobioreactor was run continuously for over one month with an average efficiency of light energy conversion to H₂ of 1.4%. Another H₂-producing approach employed a unique type of activity found in a strain of photosynthetic bacteria that shifts CO (and H₂O) into H₂ (and CO₂) in darkness. Continuous dark H₂ production byRhodobacter sp. CBS from CO (in anticipation of using synthesis gas as the future substrate) at rates up to 140 mL . g cdw^-1 . h^-1 was observed in a bubble-train bioreactor for more than 10 d.

     

Fermentation of rice straw/chicken manure to carboxylic acids using a mixed culture of marine mesophilic micoorganisms

Countercurrent fermentation of rice straw and chicken manure to carboxylic acids was performed using a mixed culture of marine mesophilic microorganisms. To increase the digestibility of the biomass, rice straw, and chicken manure were pretreated with 0.1 g Ca(OH)₂/g biomass. Fermentation was performed for 80% rice straw and 20% chicken manure at various volatile solid loading rates (VSLR) and liquid residence times (LRT). The highest acid productivity of 1.69 g/(L·d) occurred at a total acid concentration of 32.4 g/L. The highest conversion (0.69 g VS digested/g VS fed) and yield (0.29 g total acids/g VS fed) were at a total acid concentration of 25 g/L. A Continuum Particle Distribution Model of the process predicted the experimental total acid concentration and conversion results with an average error of 6.41% and 6.15%, respectively. Results show how total acid concentrations, conversions, and yields vary with VSLR and LRT in the MixAlco process.     

Characterization of a Recombinant Flocculent Saccharomyces cerevisiae Strain that Co-ferments Glucose and Xylose: I. Influence of the Ratio of Glucose/Xylose on Ethanol Production

Glucose/xylose mixtures (90 g/L total sugar) were evaluated for their effect on ethanol fermentation by a recombinant flocculent Saccharomyces cerevisiae, MA-R4. Glucose was utilized faster than xylose at any ratio of glucose/xylose, although MA-R4 can simultaneously co-ferment both sugars. A high percentage of glucose can increase cell biomass production and therefore increase the rate of glucose utilization (1.224 g glucose/g biomass/h maximum) and ethanol formation (0.493 g ethanol/g biomass/h maximum). However, the best ratio of glucose/xylose for the highest xylose consumption rate (0.209 g xylose/g biomass/h) was 2:3. Ethanol concentration and yield increased and by-product (xylitol, glycerol, and acetic acid) concentration decreased as the proportion of glucose increased. The maximum ethanol concentration was 41.6 and 21.9 g/L after 72 h of fermentation with 90 g/L glucose and 90 g/L xylose, respectively, while the ethanol yield was 0.454 and 0.335 g/g in 90 g/L glucose and 90 g/L xylose media, respectively. High ethanol yield when a high percentage of glucose is available is likely due to decreased production of by-products, such as glycerol and acetic acid. These results suggest that ethanol selectivity is increased when a higher proportion of glucose is available and reduced when a higher proportion of xylose is available.     

亚心形四爿藻培养和产氢过程一体化平板光生物反应系统

利用整合了燃料电池的平板光生物反应器, 探讨了将亚心型四爿藻高密度培养和产氢两段工艺一体化集成的可行性. 在培养阶段通入体积分数为2%~5%的CO2可使藻细胞迅速增殖, 9 d内即可达到产氢要求的生物量(8.5×106 cell/mL). 通过叶绿素荧光参数分析, 选择2%的CO2培养的藻进行后续的产氢实验. 结果表明, PSⅡ活性和光合电子传递速率均随时间的推移而逐渐下降. 通过对产氢动力学曲线的分析, 计算出最大产氢速率为1.1 mL/(h·L), 持续产氢时间为60 h.     

Performance of an internal-loop airlift bioreactor for treatment of hexane-contaminated air

Hexane is a toxic volatile organic compound that is quite abundant in gas emissions from chemical industries and printing press and painting centers, and it is necessary to treat these airstreams before they discharge into the atmosphere. This article presents a treatment for hexane-contaminated air in steady-state conditions using an internal-loop airlift bioreactor inoculated with a Pseudomonas aeruginosa strain. Bioprocesses were conducted at 20-mL/min, a load of 1.26 g/m³ of C₆H₁₄, and a temperature of 28°C. The results of hexane removal efficiencies were presented as a function of the inoculum size (approx 0.07 and 0.2 g/L) and cell reuse. Bioprocess monitoring comprises quantification of the biomass, the surface tension of the medium, and the hexane concentration in the fermentation medium as well as in the inlet and outlet airstreams. The steady-state results suggest that the variation in inoculum size from 0.07 to 0.2 g/L promotes hexane abatement from the influent from 65 to 85%, respectively. Total hydrocarbon removal from the waste gas was achieved during experiments conducted using reused cells at an initial microbial concentration of 0.2 g/L.     

Effect of Zeolite NaY and Ca-Montmorillonite on Ethanol Production Using Synthetic Molasses

The influence of Ca-Montmorillonite (Ca-MNT) and zeolite NaY addition on ethanol production from synthetic molasses by S. cerevisiae 251 TP(3-2) was studied by the measurement of biomass concentrations and metal ion concentration with respect to fermentation time. Addition of 5 g/L Ca-MNT and 10 g/L zeolite NaY resulted in an increase in both ethanol concentration and ethanol production rate. This increase was 24 and 40% for ethanol concentration and 65 and 87% for production rate, respectively. From the ion analyses, it was observed that the NaY added to the medium decreased the toxic concentration of zinc, manganese, and iron cations and acted also as a pH regulator. Ca-MNT added to the medium decreased the concentration of Na⁺ ions, which is known to have a toxic effect on glycolysis and cell concentration. These effects caused improvement in the ethanol production rate.     

Astaxanthinogenesis in the yeastPhaffia rhodozyma

Astaxanthin is a diketo-dihydroxy-carotenoid produced byPhaffia rhodozyma, a basidiomicetous yeast. A low-cost fermentation medium consisting of raw sugarcane juice and urea was developed to exploit the active sucrolytic/urelolytic enzyme apparatus inherent to the yeast. As compared to the beneficial effect of 0.1 g% urea, a ready nitrogen source, mild phosphoric pre inversion of juice sucrose to glucose and fructose, promptly fermentable carbon sources, resulted in smaller benefits. Corn steep liquor (CSL) was found to be a valuable supplement for both yeast biomass yield (9.2 g dry cells/L) and astaxanthin production (1.3 mg/g cells). Distillery effluent (vinace), despite only a slightly positive effect on yeast growth, allowed for the highest pigment productivity (1.9 mg/g cells). Trace amounts of Ni² (1 mg/L, as a cofactor for urease) resulted in controversial effects, namely, biomass decrease and astaxanthin increase, with no effect on the release (and uptake) of ammonium ion from urea. Since the synthesized astaxanthin is associated with the yeast cell and the pigment requires facilitated release for aquaculture uses (farmed fish meat staining), an investigation of the yeast cell wall was undertaken using detergent-treated cells. The composition of the rigid yeast envelope was found to be heterogeneous. Its partial acid or enzymatic depolymerization revealed glucose and xylose as common monomeric units of the cell-wall glycopolymers. Yeast cell-wall partial depolymerization with appropriate hydrolases may improve the pigment bioavailability for captive aquatic species and poultry.

     

Inhibitors Compounds on Sugarcane Bagasse Saccharification: Effects of Pretreatment Methods and Alternatives to Decrease Inhibition

Considering bioethanol production, extensive research has been performed to decrease inhibitors produced during pretreatments, to diminish energy input, and to decrease costs. In this study, sugarcane bagasse was pretreated with NaOH, H₂SO₄, and water. The higher concentration of phenols, 3.3 g/L, was observed in biomass liquid fraction after alkaline pretreatment. Acid pretreatment was responsible to release considerable acetic acid concentration, 2.3 g/L, while water-based pretreatment was the only to release formic acid, 0.02 g/L. Furans derivatives were not detected in liquid fractions regardless of pretreatment. Furthermore, washing step removed most of the phenols from pretreated sugarcane bagasse. Saccharification of alkali-pretreated biomass plus polyethylene glycol (PEG) at 0.4% (w/v) enhanced 8 and 26% the glucose and the xylose release, respectively, while polyvinylpyrrolidone (PVP) also at 0.4% (w/v) increased the release by 10 and 31% of these sugars, respectively, even without washing and filtration steps. Moreover, these polymers cause above 50% activation of endoglucanase and xylanase activities which are crucial for biomass hydrolysis.

     

Ethanol Production from Sugarcane Bagasse by Zymomonas mobilis Using Simultaneous Saccharification and Fermentation (SSF) Process

Considerable efforts have been made to utilize agricultural and forest residues as biomass feedstock for the production of second-generation bioethanol as an alternative fuel. Fermentation utilizing strains of Zymomonas mobilis and the use of simultaneous saccharification and fermentation (SSF) process has been proposed. Statistical experimental design was used to optimize the conditions of SSF, evaluating solid content, enzymatic load, and cell concentration. The optimum conditions were found to be solid content (30%), enzymatic load (25 filter paper units/g), and cell concentration (4 g/L), resulting in a maximum ethanol concentration of 60 g/L and a volumetric productivity of 1.5 g L^?1?h^?1.     

Production of Xylitol from D-Xylose byDebaryomyces hansenii

Xylitol, a naturally occurring five-carbon sugar alcohol, can be produced from D-xylose through microbial hydrogenation. Xylitol has found increasing use in the food industries, especially in confectionary. It is the only so-called “second-generation polyol sweeteners” that is allowed to have the specific health claims in some world markets. In this study, the effect of cell density on the xylitol production by the yeastDebaryomyces hansenii NRRL Υ-7426 from D-xylose under microaerobic conditions was examined. The rate of xylitol production increased with increasing yeast cell density to 3 g/L. Beyond this amount there was no increase in the xylitol production with increasing cell density. The optimal pH range for xylitol production was between 4.5 and 5.5. The optimal temperature was between 28 and 37°C, and the optimal shaking speed was 300 rpm. The rate of xylitol production increased linearly with increasing initial xylose concentration. A high concentration of xylose (279 g/L) was converted rapidly and efficiently to produce xylitol with a product concentration of 221 g/L was reached after 48 h of incubation under optimum conditions.

     

Microbial Lipid Production from Corn Stover via Mortierella isabellina

Microbial lipid is a promising source of oil to produce biofuel if it can be generated from lignocellulosic materials. Mortierella isabellina is a filamentous fungal species featuring high content of oil in its cell biomass. In this work, M. isabellina was studied for lipid production from corn stover. The experimental results showed that M. isabellina could grow on different kinds of carbon sources including xylose and acetate, and the lipid content reached to 35 % at C/N ratio of 20. With dilution, M. isabellina could endure inhibition effects by dilute acid pretreatment of corn stover (0.3 g/L furfural, 1.2 g/L HMF, and 1 g/L 4-hydroxybenozic acid) and the strain formed pellets in the cell cultivations. An integrated process was developed combining the dilute acid pretreatment, cellulase hydrolysis, and cell cultivation for M. isabellina to convert corn stover to oil containing fungal biomass. With 7.5 % pretreated biomass solid loading ratio, the final lipid yield from sugar in pretreated biomass was 40 % and the final lipid concentration of the culture reached to 6.46 g/L.     

Effect of Different Carbon Sources on Cellulase Production by Marine Strain Microbulbifer hydrolyticus IRE-31-192

Cellulase is an important enzyme that can be used to breakdown lignocellulose into glucose. Microbulbifer hydrolyticus IRE-31(ATCC 700072) is a kind of marine bacterium, which could grow in high salinity medium and has fast-strong growth ability. In this study, a novel strain was screened from Microbulbifer hydrolyticus IRE-31 through mutations to produce cellulase. The effect of different carbon sources on the growth as well as on the production of cellulase of the new strain was studied. Carboxymethyl-cellulase (CMCase) activity selected to represent cellulase was proven to be effectively promoted while xylose, galactose, and melibiose as well as glucose were used as carbon sources. When xylose and glucose were chosen to be further investigated, 472.57 U/L and 266.01 U/L CMCase activity were obtained from 30 g/L glucose and 10 g/L xylose, respectively. These results clarified the effect of different carbon sources on the production of cellulase, which laid a good foundation for the further research in the production of cellulase by marine bacteria.

     

Preliminary Research on Cd^2＋ Removal from Wastewater by Sphaerotilus Natans

Influencing factors, such as the biomass concentration, pH value, temperature and time of adsorption on the removal of Cd^2＋ were investigated when Sphaerotilus natans was used as a biosorbent. The results show that Sphaerotilus natans has a significant effect on the adsorption of Cd^2＋. The process of adsorption will reach the equilibrium in 20 min. Temperature has no significant effect on the adsorption. The removal rate of Cd^2＋. can approach to 90% under the conditions of pH 5.5, the biomass mass concentration O. 6 g/L and the initial mass concentration of cadmium ≤20 mg/L. The maximum specific uptake is 1.9 mmol/g dry biomass . The adsorption was fitted with the Freundlich isotherm adsorption equation when the concentration of Cd^2＋ is in a range of 0--100 mg/L. HCl and EDTA can desorb Cd^2＋ from biomass effectively.     

Efficient Production of a Thermophilic 2-Deoxyribose-5-Phosphate Aldolase in Glucose-Limited Fed-Batch Cultivations of <Emphasis Type="Italic">Escherichia coli</Emphasis> by Continuous Lactose Induction Strategy

The production of a thermophilic 2-deoxyribose-5-phosphate aldolases (DERA) in Escherichia coli BL21 under continuous lactose induction strategy was investigated. The process was combined with the exponential feeding method, controlling the feeding rate to maintain the specific growth rate at 0.15 h⁻¹. The results indicate that the lactose concentration in the feed medium affected directly the expression of the target protein. The use of 50 g/L in the feed medium resulted in the biomass concentration of 39.3 g DCW/L, and an expression level of above 30%, and the maximum final DERA concentration of 16,200 U/L. Furthermore, the acetate concentration remained at a low level in the fed-batch phase, less than 0.5 g/L. In conclusion, combining glucose feeding with lactose induction is a more powerful way to achieve high cell density cultures and to efficiently produce the thermophilic DERA. The results also indicate the potential industrial utility in the scale production of other recombinant proteins.     

Carob pod: A new substrate for citric acid production by Aspergillus niger

The production of citric acid from carob pod extract byA. niger in surface fermentation was investigated. A maximum citric acid concentration (85.5 g/L), citric acid productivity (4.07 g/L/d), specific citric acid production rate (0.18 g/g/d), and specific sugar uptake rate (0.358 g/g/d) was achieved at an initial sugar concentration of 200 g/L, pH of 6.5, and a temperature of 30°C. Other kinetic parameters, namely, citric acid yield, biomass yield, specific biomass production rate, and fermentation efficiency were maximum at pH 6.5, temperature 30°C, and initial sugar concentration 100 g/L. The external addition of methanol into the carob pod extract at a concentration up to 4% (v/v) improved the production of citric acid.     

Kinetics of ethanol production from carob pods extract by immobilizedSaccharomyces cerevisiae cells

Kinetics of ethanol production from carob pods extract by immobilizedS. cerevisiae cells in static and shake flask fermentation have been investigated. Shake flask fermentation proved to be a better fermentation system for the production of ethanol than static fermentation. The optimum values of ethanol concentration, ethanol productivity, ethanol yield, and fermentation efficiency were obtained at pH range 3.5–6.5 and temperature between 30–35°C. A maximum ethanol concentration (65 g/L), ethanol productivity (8.3 g/Lh), ethanol yield (0.44 g/g), and fermentation efficiency (95%) was achieved at an initial sugar concentration of 200, 150, 100, and 200 g/L, respectively. The highest values of specific ethanol production rate and specific sugar uptake rate were obtained at pH 6.5, temperature 40°C, and initial sugar concentration of 100 g/L. Other kinetic parameters, biomass concentration, biomass yield, and specific biomass production rate were maximum at pH 5.5, temperature 30°C, and initial sugar concentration 150 g/L. Under the same fermentation conditions non-sterilized carob pod extract gave higher ethanol concentration than sterilized medium. In repeated batch fermentations, the immobilizedS. cerevisiae cells in Ca-alginate beads retained their ability to produce ethanol for 5 d.     

Rhamnolipid Production by <Emphasis Type="Italic">Pseudomonas Aeruginosa</Emphasis> GIM 32 Using Different Substrates Including Molasses Distillery Wastewater

A rhamnolipid production strain newly isolated from oil-contaminated soil was identified as Pseudomonas aeruginosa GIM32 by its morphology and 16S rDNA sequence analysis. The effect of carbon source and carbon to nitrogen (C/N) ratio on rhamnolipids production was investigated. Palm oil was favorable as a carbon source for rhamnolipid production. The maximum biomass and rhamnolipid concentration were 8.24 g/L and 30.4 g/L, respectively, with an optimization medium containing 50 g/L palm oil and 5 g/L sodium nitrate. Molasses distillery wastewater as an unconventional substrate for rhamnolipid production was investigated. It was found that 2.6 g/L of rhamnolipids was produced; this amount was higher than that of past reports using wastewater as a substrate. In addition, 44% of the chemical oxygen demand of wastewater was removed at the same time under the optimization condition. Eleven kinds of different molecular weight rhamnolipid homologues were identified in the rhamnolipids obtained from molasses distillery wastewater by P. aeruginosa GIM32 by LC–MS analysis.