An optical-density-based feedback feeding method for ammonium concentration control in Spirulina platensis cultivation

Cultivation of Spirulina platensis using ammonium salts or wastewater containing ammonium as alternative nitrogen sources is considered as a commercial way to reduce the production cost. In this research, by analyzing the relationship between biomass production and ammonium- N consumption in the fed-batch culture of Spirulina platensis using ammonium bicarbonate as a nitrogen nutrient source, an online adaptive control strategy based on optical density (OD) measurements for controlling ammonium feeding was presented. The ammonium concentration was successfully controlled between the cell growth inhibitory and limiting concentrations using this OD-based feedback feeding method. As a result, the maximum biomass concentration (2.98 g/l), productivity (0.237 g/l·d), nitrogen-to-cell conversion factor (7.32 gX/gN), and contents of protein (64.1%) and chlorophyll (13.4 mg/g) obtained by using the OD-based feedback feeding method were higher than those using the constant and variable feeding methods. The OD-based feedback feeding method could be recognized as an applicable way to control ammonium feeding and a benefit for Spirulina platensis cultivations.     

Raceway cultivation of Spirulina platensis using underground water

The semi-outdoor cultivation of Spirulina platensis was attempted using an underground-water-based medium. Occurrence of contaminant organisms such as Chlorella sp. and Chlamydomonas sp. was not found from a microscopic observation and bacteria were not detected from denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rDNA during the cultivation, owing to pH control and the high quality of the underground water. The mean productivity was high at 10.5 g/m2/d with a range of 4.2-12.3 g/m2/d despite the unfavorable weather conditions of the rainy season. The cultivated S. platensis included a normal protein content of 58.9%. Consequently, the underground water improved the biomass productivity and the biomass quality because of an abundant supplementation of natural minerals and through a contaminant-free culture.     

Efficient docosahexaenoic acid production by Schizochytrium sp. via a two-phase pH control strategy using ammonia and citric acid as pH regulators

The cell growth and docosahexaenoic acid (DHA) synthesis of Schizochytrium sp. are closely related to the culture pH. A two-phase pH control strategy based on nitrogen consumption was developed in which pH 7.0 was used for biomass accumulation and pH 5.0 for DHA synthesis. Using this strategy, the cell dry weight and DHA content reached 98.07 and 25.85 g/L, respectively. Furthermore, ammonia and citric acid were used as pH regulators. Application of citric acid further resulted in 7.88 and 4.87% improvements of total lipids and the ratio of DHA to total fatty acids, respectively. Ammonia, as a suitable nitrogen source, promoted non-lipid biomass accumulation. Using this method, a maximum DHA yield of 32.75 g/L was obtained with non-lipid biomass (58.01 g/L) and the ratio of DHA to total fatty acids (52.36%). This study provides an easy strategy for large-scale industrial production of DHA via high-cell-density fermentation of Schizochytrium sp.     

Production of biomass and nutraceutical compounds by Spirulina platensis under different temperature and nitrogen regimes

The cyanobacterium Spirulina platensis has been used by humans because of its nutritional and possibly medicinal effects. Our study evaluated the influence of temperature and nitrogen concentration in the medium on the production of biomass by this cyanobacterium and the biomass composition in protein, lipid and phenolic compounds. We found that at 35 degrees C there was a negative effect on biomass production but a positive effect on the production of protein, lipids and phenolics, the highest levels of these compounds being obtained in Zarrouk's medium containing 1.875 or 2.500 g l(-1) sodium nitrate. Higher biomass densities and productivity were obtained at 30 degrees C than at 35 degrees C, but nitrogen concentration appeared to have no effect on the amount of protein, lipid or phenolics, indicating that at 30 degrees C the concentration of sodium nitrate in Zarrouk's medium (2.50 g l(-1)) can be reduced without loss of productivity, an important cost-saving factor in large-scale cultivation.     

Thermal characteristics of Spirulina platensis cells under nongrowing conditions at various values of pH medium

The total value of heat (-Q) evolved by green-blue microalgae Spirulina platensis cells in a dark and stationary regime in the range of pH values 8.0-11.6 was determined. It was established that (-Q) reaches its maximum value at 360 +/- 40 J/g of dry biomass in the pH range 9.3-10.3 and then sharply dropped relative to these values and reached zero at pH 7.5 +/- 0.2 and 11.8 +/- 0.2. It is affirmed that an optimum regime for preservation of Spirulina platensis cell viability in a dark and stationary regime is pH range 9.3-10.3. It was also shown that the peak of heat evolution with maximum about 45 degrees C, reflecting mainly the respiration of cells (oxygen absorption rate), did not displace along the temperature scale at a change of pH from 9.3 to 10.4 and slightly displaced lower and higher of these values of pH. It is supposed that the thermostability of biomacromolecules and their complexes responsible for cell respiration does not depend on pH medium in pH range 9.3-10.3.     

Comparison of batch and semicontinuous cultures for production of protein from mesquite wood by Brevibacterium sp. JM98A

The production of protein by a Brevibacterium sp. JM98A usingmesquite wood as the substrate was compared in batch and semicontinuous cultures. A 14 liter glass fermentor with automatic pH, temperature, and foam control was used for the study. A pH range of 6.6 to 7.2 was optimum for the growth of JM98A. The batch and semicontinuous cultures were compared on the basis of viable cell counts, protein production, CMC-Ase (β-1,4-glucanase) activity, and filter paper cellulase (β-1,4-glucan cellobiohydrolyase) activity. Total hexose, cellulose, and reducing sugar consumption were measured. The semicontinuous process yielded 2.97 times as much protein in 72 hr as the batch cultures. Most of the biomass resulted from the utilization of soluble sugars rather than from the degradation of cellulose during the semicontinuous process.     

Quantitative physiology of Penicillium cyclopium grown on whey for production of microbial protein

Summary A filamentous fungus Penicillium cyclopium, capable of growing on deproteinized whey was isolated and characterized for the purpose of production of microbial protein.This organism has a maximum specific growth rate of 0.2 h^–1 at pH 3.0 to 4.5 and 28°C in a medium containing only ammonium nitrogen and deproteinized whey. The yield coefficients are 0.68 g biomass/g lactose, 12.0 g biomass/g nitrogen, and 2.10 g biomass/g oxygen, respectively.Crude protein and total nucleic acid contents of this organism are 47.5% and 7.4% (dry cell weight basis), respectively. The profile of essential amino acids shows that it could be a good source of animal feed or food protein.     

A one-step process for the production of single-cell protein and amyloglucosidase

Summary A new Rhizopus species was isolated from traditional Indonesian food, tempeh. The newly isolated species was similar in its morphological characteristics to Rhizopus oligosporus UQM 145F, but grew faster on potato-dextrose agar as well as in submerged culture. The new isolate was found to convert ground cassava tuber directly into single cell protein without pretreatment due to its high amyloglucosidase formation.From 100 g ground tuber, a dry biomass of 33.75 g containing 26.48% true protein together with 60 ml of highly active amyloglucosidase (282 units) was obtained in 12 h. The amyloglucosidase was recovered by ultrafiltration, releasing 26.226 millimol glucose/l/min from soluble starch. The crude enzyme exhibited a pH optimum between 4.6 and 5.0, a temperature optimum between 55 and 60° C and an apparent Km of 3.125 g/l. High substrate concentrations and ammonium sulphate are inhibitory to the enzyme.     

High cell density cultivation of a novel Aurantiochytrium sp. strain TC 20 in a fed-batch system using glycerol to produce feedstock for biodiesel and omega-3 oils

A recently isolated Australian Aurantiochytrium sp. strain TC 20 was investigated using small-scale (2 L) bioreactors for the potential of co-producing biodiesel and high-value omega-3 long-chain polyunsaturated fatty acids. Higher initial glucose concentration (100 g/L compared to 40 g/L) did not result in markedly different biomass (48 g/L) or fatty acid (12–14 g/L) yields by 69 h. This comparison suggests factors other than carbon source were limiting biomass production. The effect of both glucose and glycerol as carbon sources for Aurantiochytrium sp. strain TC 20 was evaluated in a fed-batch process. Both glucose and glycerol resulted in similar biomass yields (57 and 56 g/L, respectively) by 69 h. The agro-industrial waste from biodiesel production—glycerol—is a suitable carbon source for Aurantiochytrium sp. strain TC 20. Approximately half the fatty acids from Aurantiochytrium sp. strain TC 20 are suitable for development of sustainable, low emission sources of transportation fuels and bioproducts. To further improve biomass and oil production, fortification of the feed with additional nutrients (nitrogen sources, trace metals and vitamins) improved the biomass yield from 56 g/L (34 % total fatty acids) to 71 g/L (52 % total fatty acids, cell dry weight) at 69 h; these yields are to our knowledge around 70 % of the biomass yields achieved, however, in less than half of the time by other researchers using glycerol and markedly greater than achieved using other industrial wastes. The fast growth and suitable fatty acid profile of this newly isolated Aurantiochytrium sp. strain TC 20 highlights the potential of co-producing the drop-in biodiesel and high value omega-3 oils.     

光质对工程聚球藻7002生长及psbA启动子的调控

工程聚球藻的psbA基因及其所用载体的启动子Ppsb A均受光质调控,利用该机制可通过光质调控提高聚球藻的光合效率及其外源基因表达率。以转vp28基因聚球藻7002为实验材料,通过优化光强、温度及pH,解除光限制因素并提高光能利用率。通过改变白光、红光及蓝光的比例,调控光质组成及单色光的光强,检测细胞生长、外源基因的表达及psb A基因的转录。研究结果表明:高比例蓝光下,vp28基因的表达率达到2.4%,是纯白光下的3倍,重组蛋白VP28的积累量提高至2倍。高比例红光抑制了psb AII、psb AIII基因及外源基因的表达,但促使生物量在3 d内突破1.5 g/L。本研究为蓝藻的生物制药和工程蓝藻代谢产物的产业化提供了理论基础。     

Isomaltulose production using free cells: optimisation of a culture medium containing agricultural wastes and conversion in repeated-batch processes

The enzyme glucosyltransferase is an industrially important enzyme since it produces non-cariogenic isomaltulose (6-O-alpha-D-glucopyronosyl-1-6-D-fructofuranose) from sucrose by intramolecular transglucosylation. The experimental designs and response surface methodology (RSM) were applied for the optimisation of the nutrient concentrations in the culture medium for the production of glucosyltransferase by Erwinia sp. D12 in shaken flasks at 200 rpm and 30 degrees C. A statistical analysis of the results showed that, in the range studied, the factors had a significant effect (P < 0.05) on glucosyltransferase production and the highest enzyme activity (10.84 U/ml) was observed in culture medium containing sugar cane molasses (150 g l(-1)), corn steep liquor (20 g l(-1)), yeast extract Prodex Lac SD (15 g l(-1)) and K2HPO4 (0.5 g l(-1)) after 8 h at 30 degrees C. The production of cell biomass by the strain of Erwinia sp. D12 was carried out in a 6.6-l fermenter with a mixing rate of 200 rpm and an aeration rate of 1 vvm. Fermentation time, cellular growth, medium pH and glucosyltransferase production were observed. The greatest glucosyltransferase activity was 22.49 U/ml, obtained after 8 h of fermentation. The isomaltulose production from sucrose was performed using free Erwinia sp. D12 cells in a batch process using an orbital shaker. The influence of the parameters sucrose concentration, temperature, pH, and cell concentration on the conversion of sucrose into isomaltulose was studied. The free cells showed a high conversion rate of sucrose into isomaltulose using batch fermentation, obtaining an isomaltulose yield of 72.11% from sucrose solution 35% at 35 degrees C.     

Simultaneous single-cell protein production and COD removal with characterization of residual protein and intermediate metabolites during whey fermentation by K. marxianus

Cheese whey fermentation with Kluyveromyces marxianus was carried out at 40 °C and pH 3.5 to examine simultaneous single-cell protein production and chemical oxygen demand (COD) removal, determine the fate of soluble whey protein and characterize intermediate metabolites. After 36 h of batch fermentation, the biomass concentration increased from 2.0 to 6.0 g/L with 55 % COD reduction (including protein), whereas soluble whey protein concentration decreased from 5.6 to 4.1 g/L. It was confirmed through electrophoresis (SDS-PAGE) that the fermented whey protein was different from native whey protein. HPLC and GC–MS analysis revealed a change in composition of organic compounds post-fermentation. High inoculum concentration in batch fermentation resulted in an increase in biomass concentration from 10.3 to 15.9 g/L with 80 % COD reduction (including protein) within 36 h with residual protein concentration of 4.5 g/L. In third batch fermentation, the biomass concentration increased from 7.3 to 12.4 g/L with 71 % of COD removal and residual protein concentration of 4.3 g/L after 22 h. After 22 h, the batch process was shifted to a continuous process with cell recycle, and the steady state was achieved after another 60 h with biomass yield of 0.19 g biomass/g lactose and productivity of 0.26 g/L h. COD removal efficiency was 78–79 % with residual protein concentration of 3.8–4.2 g/L. The aerobic continuous fermentation process with cell recycle could be applied to single-cell protein production with substantial COD removal at low pH and high temperature from cheese whey.     

Zinc removal from model solution and wastewater by Arthrospira (Spirulina) Platensis biomass

The biosorption of zinc from model solution as well as wastewater by Arthrospira (Spirulina) platensis biomass was studied. Adsorption capacity of the biosorbent was investigated as a function of contact time between adsorbent and zinc, the initial metals and sorbent concentration, pH value, and temperature. The ability of Arthrospira biomass for zinc biosorption exhibited a maximum at the pH range 4–8. Equilibrium data fitted well with the Langmuir model as well as the Freundlich model with maximum adsorption capacity of 7.1 mg/g. The pseudo second-order model was found to correlate well with the experimental data. Different thermodynamic parameters, ΔG°, ΔH° and ΔS° were evaluated and it has been found that the sorption was feasible, spontaneous, and endothermic in nature. The process of zinc removal from industrial effluent was studied at different time of sorbat–sorbent interaction and different sorbent dosage. Maximum zinc removal (83%) was obtained at sorbent concentration 60 g/L during 1-h experiment. The results indicate that Arthrospira platensis biomass could be effectively used for zinc removal from industrial effluents.     

Influence of specific growth rate on biomass yield, productivity, and compostion of Candida utilis in batch and continuous culture.

Candida utilis was grown in batch and continuous culture on prickly pear juice as sole carbon and energy source. In batch culture the maximum specific growth rate (mum) and the substrate yield coefficient (Yps) varied according to sugar concentration. When the fermentation was carried out with 1% sugar, mum and Ys were 0.47/h and 42.6%, respectively. The best yields occurred in a chemostat at the pH range of 3.5 to 4.5 and temperature of 30 C. A beneficial effect on Ys was observed when the dilution rate (D) was increased. At a D of 0.55/h, the productivity was 2.38 g/liter per h. The maintenance coefficient attained a value of 0.09 g of sugar/g of biomass per h. Increases of D produced higher protein contents of the biomass. The information obtained indicates that protein production with Candida utilis, using prickly pear juice, should be carried out a high dilution rates where the Ys and protein content of the cell mass are also higher.     

Biofixation of carbon dioxide by Spirulina sp. and Scenedesmus obliquus cultivated in a three-stage serial tubular photobioreactor

The increase in the concentration of atmospheric carbon dioxide is considered to be one of the main causes of global warming. As estimated by the Intergovernmental Panel on Climate Change (IPCC) criteria, about 10-15% of the gases emitted from the combustion coal being in the form of carbon dioxide. Microalgae and cyanobacteria can contribute to the reduction of atmospheric carbon dioxide by using this gas as carbon source. We cultivated the Scenedesmus obliquus and Spirulina sp. at 30 degrees C in a temperature-controlled three-stage serial tubular photobioreactor and determined the resistance of these organisms to limitation and excess of carbon dioxide and the capacity of the system to fix this greenhouse gas. After 5 days of cultivation under conditions of carbon limitation both organisms showed cell death. Spirulina sp. presenting better results for all parameters than S. obliquus. For Spirulina sp. the maximum specific growth rate and maximum productivity was 0.44 d(-1), 0.22 g L(-1)d(-1), both with 6% (v/v) carbon dioxide and maximum cellular concentration was 3.50 g L(-1) with 12% (v/v) carbon dioxide. Maximum daily carbon dioxide biofixation was 53.29% for 6% (v/v) carbon dioxide and 45.61% for 12% carbon dioxide to Spirulina sp. corresponding values for S. obliquus being 28.08% for 6% (v/v) carbon dioxide and 13.56% for 12% (v/v) carbon dioxide. The highest mean carbon dioxide fixation rates value was 37.9% to Spirulina sp. in the 6% carbon dioxide runs.     

Use of the effluent from biogas production for cultivation of Spirulina

The effluent from the biogas process was tested as a nutrient source during cultivation of the protein-rich and edible microalgae Spirulina (Arthrospira platensis) and compared with conventional Spirulina medium. Equal biomass production was observed until late exponential phase and no significant differences could be observed between the treatments in protein amount, amino acid composition, and total lipid concentration. The concentration of the pigment phycocyanin differed significantly between Spirulina medium and the effluent-based medium (63.3 ± 11.7 and 86.2 ± 1.9 mg g⁻¹, respectively). Slightly higher concentrations of saturated fatty acids, mainly palmitic acid, were observed in the biomass produced in Spirulina medium than in that produced in the effluent-based medium. In the biomass produced in the effluent-based medium, the cadmium concentration was 0.07 ± 0.05 mg kg⁻¹ of dry weight, whereas it was below the detection limit in the biomass produced in Spirulina medium. There is a need to identify new food and feed resources and a possible future scenario is to integrate Spirulina production into the biogas plant for protein production as it contains more than 60% of protein on dry weight basis. In that scenario, it is important to control heavy metal concentrations in the biogas slurry fed to Spirulina.

     

Influence of specific growth rate on biomass yield, productivity, and compostion of Candida utilis in batch and continuous culture.

Candida utilis was grown in batch and continuous culture on prickly pear juice as sole carbon and energy source. In batch culture the maximum specific growth rate (mum) and the substrate yield coefficient (Yps) varied according to sugar concentration. When the fermentation was carried out with 1% sugar, mum and Ys were 0.47/h and 42.6%, respectively. The best yields occurred in a chemostat at the pH range of 3.5 to 4.5 and temperature of 30 C. A beneficial effect on Ys was observed when the dilution rate (D) was increased. At a D of 0.55/h, the productivity was 2.38 g/liter per h. The maintenance coefficient attained a value of 0.09 g of sugar/g of biomass per h. Increases of D produced higher protein contents of the biomass. The information obtained indicates that protein production with Candida utilis, using prickly pear juice, should be carried out a high dilution rates where the Ys and protein content of the cell mass are also higher.     

pH对小球藻Chlorella sp. XQ-200419光合作用、生长和产油的影响

以一种生长快、油脂含量高的小球藻(Chlorella sp. XQ-200419)为实验材料, 利用测定净光合放氧速率的方法研究了pH对其光合作用的影响; 使用改良的BG-11培养基在微藻环形培养池模拟系统中进行分批培养, 培养周期为8d, 培养过程中使用 pH控制仪在线监测藻液的pH, 根据pH变化, 自动接通、关闭CO2通气管道, 将藻液pH分别控制在5.06.0, 7.08.0, 8.09.0, 9.010.0, 10.011.0内, 研究pH对生长速率、生物质面积产率、总脂含量和总脂面积产率的影响。主要结果如下: 藻液pH对小球藻Chlorella sp. XQ-200419光合放氧、生长速率、生物质产率、总脂含量和产率都有显著影响, 适宜的pH范围是7.09.0, 在此范围内, 光合放氧、生长速率、生物质产率、总脂含量和产率均保持较高水平, 且pH的影响不显著; pH低于7.0, 高于9.0, 其光合放氧、生长速率、生物质产率、总脂含量和产率都显著降低。这表明pH对小球藻Chlorella sp. XQ-200419光合作用的影响和对生长、产油的影响是一致的。pH 7.08.0, 小球藻的生物质平均面积产率和总脂平均面积产率都达到最大值, 分别是8.9 g/(m2d)和2269.5 mg/(m2d); 当藻液pH超过10.0, 生物质平均面积产率和总脂平均面积产率分别降低42.1%和60.0%。适合于小球藻生长的pH也有利于其积累油脂, 所以, pH对小球藻产油的影响是一种适宜模式, 而非胁迫模式。规模化培养小球藻Chlorella sp. XQ-200419, 通过补充CO2将藻液pH控制在7.09.0内, 可以获得高生物质产率和总脂产率。研究结果反映出pH对小球藻光合作用、生长和产油影响的规律, 也为规模化培养小球藻生产微藻油脂过程中合理控制藻液pH提供了依据。       

Growth and Polysaccharide Production by Methylocystis parvus OBBP on Methanol

Methylocystis parvus OBBP, an obligate methylotroph originally isolated as a methane-utilizing bacterium, was cultivated on methanol as a sole source of carbon. After adaptation to high methanol levels, this organism grew on methanol with a maximum specific growth rate of 0.65 h. The pH optimum for growth was between 7 and 9, and the temperature optimum was between 30 and 37 degrees C. Methanol concentrations higher than 5% (by weight) were toxic. Formaldehyde, at a concentration greater than 1 mM, inhibited growth. Formate was neither a substrate nor an inhibitor. An extracellular viscous heteropolysaccharide was produced during growth. The maximum production of the total biomass was 14.5 g (dry weight) per liter of broth. The dried biomass contained 22% (wt/wt) crude protein and 62% (wt/wt) polysaccharide. The main components of the polysaccharide were d-glucose (82%) and l-rhamnose (14%).     

Effects of Temperature and pH on Cell Growth of a Flocculating Microalga (Chlorella vulgaris) JSC-7

微藻是可广泛用于健康食品及水产养殖的饵料,同时,微藻细胞内积累的油脂可作为可再生生物燃料,因此微藻的生长和代谢受到广泛关注。温度和pH对微藻的生物量积累有很大影响,考察不同温度和pH条件下微藻细胞的生长有助于寻找最佳的条件进行微藻的培养。自絮凝小球藻JSC-7（Chlorella vulgaris JSC-7）可实现自沉降采收,有利于降低微藻生产成本,优化其生长条件对更好地利用该微藻具有重要意义。考察了温度（22∽40℃）及pH（6.0∽10.0）对其细胞生长、叶绿素含量和油脂产量的影响。在所选取的温度及pH范围内,JSC-7细胞均可生长,显示该藻种可以适应广泛的温度和pH条件。适合细胞生长的温度依次为31℃〉28℃〉35℃〉25℃,pH依次为7.0〉8.0〉6.0。pH 8.0时生物量和油脂的积累量最多,说明该藻株在弱碱条件下更适合生长和产油。当温度为31℃、pH为7.0时,可获得最高的生长量（OD690=0.941）、叶绿素含量（19 mg/L）及油脂产量（39.07%/克干重）。