Photoautotrophic cultivating options of freshwater green microalgal Chlorococcum humicola for biomass and carotenoid production

Photoautotrophic cultivation of Chlorococcum humicola was performed in batch and continuous modes in different cultivating system arrangements to compare biomass and carotenoids’ concentration and their productivities. Batch result from stirred tank and airlift photobioreactors indicated the positive effect of increasing light intensity on growth and carotenoid production, whereas the finding from continuous cultivation indicated that carotenoid enhancement preferred high light intensity and nitrogen-deficient environment. The highest biomass (1.31?±?0.04?g?L^?1) and carotenoid (4.59?±?0.06?mg?L^?1) concentration as well as the highest productivities, 0.46?g?L^?1 d^?1 for biomass and 1.61?mg?L^?1 d^?1 for carotenoids, were obtained when maintaining high light intensity of 10 klx, BG-11 medium and 2% (v/v) CO₂ simultaneously, while the highest carotenoid content (4.84?mg?g^?1) was associated with high light intensity and nitrogen-deficient environment, which was induced by feed-modified BG-11 growth medium containing nitrate 20 folds lower than the original medium. Finally, the cultivating system arranged into smaller stirred tank photobioreactors in series yielded approximately 2.5 folds increase in both biomass and carotenoid productivities relative to using single airlift photobioreactor with equivalent working volume and similar operating condition.     

Synechococcus sp. (PTCC 6021) cultivation under different light irradiances—Modeling of growth rate-light response

Synechococcus sp. (PTCC 6021), a cyanobacterium species, was cultivated in an internally illuminated photobioreactor. The reactor was designed to achieve a monoseptic cultivation of the species. The goal was to study the growth–irradiance behavior of Synechococcus sp. (PTCC 6021). To accomplish this, different initial light irradiances were implemented inside the photobioreactor and the growth of the cells was monitored. It was observed that cell growth increased with higher light intensity until the photoinhibition occurrence at light irradiance higher than 250?μE?m^?2?s^?1. The maximum OD₆₀₀, maximum growth rate, and biomass productivity increased, and hence the extinction coefficient decreased, with the increase in light irradiance before photoinhibition. The maximum optical density (OD₆₀₀) of 5.91 was obtained with irradiance below 250?μE?m^?2?s^?1 during a growth period of 80 days. The modified Monod function could model the growth–irradiance of cells with satisfactory agreement with the experimental data. The comparison of growth–irradiance of the studied species with other photosynthetic organisms showed the same trend as for cyanobacteria with photoinhibition.     

Design of an efficient fermentation process for the production of Metarhizium acridum blastospores

Using a sequential approach, we described efficient blastospore production in a stirred tank bioreactor (3?L capacity). We used the response surface methodology to optimise the media ingredients and fermentation parameters to obtain the maximum production of blastospores by a locally collected isolate of Metarhizium acridum (Ascomycota: Hypocreales). The results showed that a liquid culture medium supplemented with monopotassium phosphate (15.17?g/L), corn steep liquor (69.25?g/L), and casamino acids (80.68?g/L) in a stirred tank bioreactor under operating conditions constant at 635?rpm, a temperature of 26°C, and pH 3.3 produced 1.25?×?10⁸?blastospores (bls)/ml, with 93% viability after 120?h of fermentation. This bioreactor yield compares favourably with the yields obtained by shake flask production and confirms the suitability of the media and production parameters for the potential scale-up fermentation production of M. acridum.     

Optimization of growth conditions and fatty acid analysis for three freshwater diatom isolates

Diatoms are a group of highly abundant and diverse aquatic algae species. They contain high lipid content along with many bioactive compounds that can be exploited for biotechnological applications. Despite these attractive attributes, diatoms are underrepresented in production projects due to difficulties in their cultivation. To optimize the growth of three freshwater diatom isolates, Cyclotella sp., Synedra sp. and Navicula sp., an orthogonal assay on N, P, Si and Fe, as well as temperature and pH, was designed using traditional single‐factor tests. We also studied the effect of using nanosilica as an alternate Si source on growth and found that the diatom isolates studied achieved their highest growth rates under different combinations of nutrient and environmental conditions. Silica had the greatest influence on growth, followed by phosphate and iron. The optimized growth conditions for Synedra sp. were N: 30 mg L^?1, P: 3 mg L^?1, Si: 14.8 mg L^?1, Fe: 0.448 mg L^?1, temperature 25°C and pH 8. For Navicula sp.: N: 20 mg L^?1, P: 2.5 mg L^?1, Si: 19.7 mg L^?1, Fe: 0.112 mg L^?1, temperature 30°C and pH 7.5–8. For Cyclotella sp.: N: 20 mg L^?1, P: 2.5 mg L^?1, Si: 19.7 mg L^?1, Fe: 0.448 mg L^?1, temperature 30°C and pH 7.5–8. Nano silica negatively affected growth in Navicula sp. and Cyclotella sp., but no such effect was observed in Synedra sp. Fatty acid profiling showed C16:0, C16:1(n ? 7), C18:0 and C20:5(n ? 3) as major fatty acids, with no significant differences in fatty acid methyl ester profiles between traditional and modified media. This work gives us a new insight into the growth requirements of freshwater diatom species, which are less studied than marine species.     

Effect of dilution rate on eicosapentaenoic acid productivity ofPhaeodactylum tricornutum utex 640 in outdoor chemostat culture

Summary Eicosapentaenoic acid (EPA) volumetric productivity from an outdoor chemostat culture ofPhaeodactylum tricornutum UTEX 640 in a 50-l tubular photobioreactor varies with dilution rate, reaching a maximum of 47.8 mg l^–1 d^–1 at D=0.36 d^–1. Continuous culture at high dilution rates' is proposed as the most adequate operating mode to maximize polyunsaturated fatty acid production.     

Effects of nitrogen delivery on chitin nanofiber production during batch cultivation of the diatom <Emphasis Type="Italic">Cyclotella</Emphasis> sp. in a bubble column photobioreactor

The photosynthetic diatom Cyclotella sp. extrudes chitin nanofibers following cell division. This diatom requires silicon for cell wall biosynthesis and division, as well as nitrogen for biosynthesis of intracellular material and extracellular chitin, an N-acetyl glucosamine biopolymer. The initial nitrogen/silicon molar ratio was the critical parameter for assessing the limits of nitrogen delivery on cell number and chitin production during batch cultivation of Cyclotella in a bubble column photobioreactor under silicon-limited growth conditions, using nitrate as the nitrogen source. The peak rate of volumetric chitin production increased linearly, from 3.0 to 46 mg chitin L^?1 day^?1, with increasing N/Si ratio over the range studied (0.82 to 8.6 mol N mol^?1 Si). However, the cell number yield and the chitin yield per cell increased asymptotically with increasing N/Si ratio, achieving a final cell number yield of 5.3?×?10⁹?±?2.6?×?10⁸ cells mol^?1 Si and chitin yield of 28.7?±?1.2 mg chitin per 10⁹ cells (1.0 S.E.). An N/Si ratio of at least 4.0 mol N mol^?1 Si achieved 90% of the asymptotic chitin yield. This study has shown that scalable cultivation systems for maximizing chitin nanofiber production will require delivery of both silicon and optimal nitrogen under silicon-limiting growth conditions to promote cell division and subsequent chitin formation.     

Optimization of exopolysaccharide production from Armillaria mellea in submerged cultures

Aims: To study the optimization of submerged culture conditions for exopolysaccharide (EPS) production by Armillaria mellea in shake‐flask cultures and also to evaluate the performance of an optimized culture medium in a 5‐l stirred tank fermenter. Methods and Results: Shake flask cultures for EPS optimal nutritional production contained having the following composition (in g l^?1): glucose 40, yeast extract 3, KH₂PO₄ 4 and MgSO₄ 2 at an optimal temperature of 22°C and an initial of pH 4·0. The optimal culture medium was then cultivated in a 5‐l stirred tank fermenter at 1 vvm (volume of aeration per volume of bioreactor per min) aeration rate, 150 rev min^?1 agitation speed, controlled pH 4·0 and 22°C. In the optimal culture medium, the maximum EPS production in a 5‐l stirred tank fermenter was 588 mg l^?1, c. twice as great as that in the basal medium. The maximum productivity for EPS (Q_p) and product yield (Y_P/S) were 42·02 mg l^?1 d^?1 and 26·89 mg g^?1, respectively. Conclusions: The optimal culture conditions we proposed in this study enhanced the EPS production of A. mellea from submerged cultures. Significance and Impact of the Study: The optimal culturing conditions we have found will be a suitable starting point for a scale‐up of the fermentation process, helping to develop the production of related medicines and health foods from A. mellea.     

Biomass and lipid enhancement in Ankistrodesmus sp. cultured with reused and minimal nutrients media

Microalgae are a promising feedstock for biofuel production. Lipid content in microalgae could be enhanced under nutrient depletion. This work investigated the effect of the nutrient on lipid accumulation in Ankistrodesmus sp. culture. Batch cultures were carried out using fresh BG11 medium, and after the harvest, the medium was reused for the next culture; this method was repeated two times. The maximum lipid productivity of 29.75 mg L^?1 day^?1 was obtained from the culture with the second reuse medium. In continuous cultures, Ankistrodesmus sp. was cultured in both fresh and modified BG11 mediums. The modified BG11 medium was adjusted to resemble the content of the first reuse medium. As a comparison between batch and continuous cultures, it was proven that the productivity in the continuous culture was better than in the batch, where the achievable maximum biomass and lipid were 188.30 and 38.32 mg L^?1 day^?1. The maximum lipid content of 34.22% was obtained from the continuous culture at a dilution rate of 0.08 day^?1, whereas the maximum saturated and unsaturated fatty acid productivities of 79.96 and 104.54 mg L^?1 day^?1 were obtained at a dilution rate of 0.16 day^?1.     

Continuous microalgae cultivation in a photobioreactor

New biomass sources for alternative fuels has become a subject of increasing importance as the nation strives to resolve the economic and strategic impacts of limited fossil fuel resources on our national security, environment, and global climate. Algae are among the most promising non‐food‐crop‐based biomass feedstocks. However, there are currently no commercially viable microalgae‐based production systems for biofuel production that have been developed, as limitations include less‐than optimal oil content, growth rates, and cultivation techniques. While batch studies are critical for determining basic growth phases and characteristics of the algal species, steady‐state studies are necessary to better understand and measure the specific growth parameters. This study evaluated the effects of dilution rate on microalgal biomass productivity, lipid content, and fatty acid profile under steady‐state conditions with continuous illumination and carbon dioxide supplemention for two types of algae. Continuous cultures were conducted for more that 3 months. Our results show that the productivity of Chlorella minutissima varied from 39 to 137 mg/L/day (dry mass) when the dilution rate varied from 0.08 to 0.64 day^?1. The biomass productivity of C. minutissima reached a maximum value (137 mg/L/day) at a dilution rate of 0.33 day^?1, while the productivity of Dunaliella tertiolecta varied from 46 to 91 mg/L/day at a dilution rate of 0.17 to 0.74 day^?1. The biomass productivity of D. tertiolecta reached a maximum value of 91 mg/L/day at a dilution rate of 0.42 day^?1. Moreover, the lipid content had no significant change with various dilution rates. Biotechnol. Bioeng. 2012; 109: 2468–2474. © 2012 Wiley Periodicals, Inc.     

EXTREMELY HIGH GROWTH RATE OF THE SMALL DIATOM CHAETOCEROS SALSUGINEUM ISOLATED FROM AN ESTUARY IN THE EASTERN SETO INLAND SEA,JAPAN1

Small single‐celled Chaetoceros sp. are often widely distributed, but frequently overlooked. An estuarine diatom with an extremely high growth potential under optimal conditions was isolated from the Shinkawa‐Kasugagawa estuary in the eastern part of the Seto Inland Sea, western Japan. It was identified as Chaetoceros salsugineum based on morphological observations. This strain had a specific growth rate of 0.54 h^?1 at 30°C under 700 μmol · m^?2 · s^?1 (about 30% of natural maximal summer light) with a 14:10 L:D cycle; there was little growth in the dark. However, under continuous light it grew at only 0.35 h^?1 or a daily specific growth rate of 8.4 d^?1. In addition, cell density, chlorophyll a, and particulate organic carbon concentrations increased by about 1000 times in 24 h at 30°C under 700 μmol · m^?2 · s^?1 with a 14:10 L:D cycle, showing a growth rate of close to 7 d^?1. This very rapid growth rate may be the result of adaptation to this estuarine environment with high light and temperature. Thus, C. salsugineum can be an important primary producer in this estuary in summer and also an important organism for further physiological and genetic research.     

Growth and biomass production with enhanced β‐glucan and dietary fibre contents of Ganoderma australe ATHUM 4345 in a batch‐stirred tank bioreactor

In this study we maximized biomass production by the basidiomycete Ganoderma australe ATHUM 4345, a species of pharmaceutical interest as it is a valuable source of nutraceuticals, including dietary fibers and glucans. We used the Biolog FF MicroPlate to screen 95 different carbon sources for growth monitoring. The pattern of substrate catabolism forms a substrate assimilation fingerprint, which is useful in selecting components for media optimization of maximum biomass production. Response surface methodology, based on the central composite design was applied to explore the optimum concentrations of carbon and nitrogen sources of culture medium in shake flask cultures. When the improved culture medium was tested in a 20‐L stirred tank bioreactor, using 13.7 g/L glucose and 30.0 g/L yeast extract, high biomass yields (10.1±0.4 g/L) and productivity of 0.09 g L^?1 h^?1 were obtained. The yield coefficients for total glucan and dietary fibers on biomass formed were 94.82±6 and 341.15±12.3 mg/g mycelium dry weight, respectively.     

Maximizing the productivity of the microalgae <Emphasis Type="Italic">Scenedesmus</Emphasis> AMDD cultivated in a continuous photobioreactor using an online flow rate control

In this study, production of the microalga Scenedesmus AMDD in a 300 L continuous flow photobioreactor was maximized using an online flow (dilution rate) control algorithm. To enable online control, biomass concentration was estimated in real time by measuring chlorophyll-related culture fluorescence. A simple microalgae growth model was developed and used to solve the optimization problem aimed at maximizing the photobioreactor productivity. When optimally controlled, Scenedesmus AMDD culture demonstrated an average volumetric biomass productivity of 0.11 g L^?1 d^?1 over a 25 day cultivation period, equivalent to a 70 % performance improvement compared to the same photobioreactor operated as a turbidostat. The proposed approach for optimizing photobioreactor flow can be adapted to a broad range of microalgae cultivation systems.     

Scale up production of isoflavonoids in cell suspension cultures of Pueraria tuberosa grown in shake flasks and bioreactor

Cell cultures of Pueraria tuberosa were grown in vessels of different sizes and 2L stirred tank bioreactor containing modified MS medium with morphactin (0.1 mg l^?1) and 2iP (5.0 mg l^?1) and 20% inoculum. Stable growth and total isoflavonoid yield of 76.6 mg l^?1 were recorded in the cultures during scale up. This was in concordance with the persistent yield of the individual isoflavonoids regardless of the vessel size.     

Treatment of high-strength purified terephthalic acid (PTA) wastewater using a magnetic porous ceramic support in laboratory-scale anaerobic-aerobic fluidized bed reactors

Abstract

Laboratory-scale anaerobic-aerobic fluidized-bed bioreactors (FBR) with porous magnetic ceramics as support were successfully applied to treat purified terephthalic acid (PTA) wastewater. After a short 14-day start-up period, the system was stably operating. During the 40?d stable period, the system organic loading rate (OLR) increased from 6.68 to 23.87?kg chemical oxygen demand (COD)/(m³d), the effluent COD and PTA were below 90 and 30?mg/L, respectively. The FBR presented excellent COD and PTA removal efficiency with a low hydraulic retention time (HRT) value of six hours. The growth kinetic parameters suggested that the biomass in FBR possess high maximum specific growth rate (μ_max?=?2.22?d^?1) and good tolerance to varied OLR (K_s?=?258.67?mg COD/L).     

Kinetics of growth, oxygen uptake, and substrate utilization of wild type and genetically engineeredBurkholderia sp

The gene (vgb) encoding the hemoglobin (VHb) ofVitreoscilla sp. was cloned intoBurkholderia sp. and the effect of VHb on the growth characteristics of genetically engineeredBurkholderia (YV1) were compared with wild typeBurkholderia (R34) using continuous flow reactors (chemostat) at various dilution rates under aerobic conditions. Batch oxygen uptake rate showed that YV1 has much higher oxygen uptake rate than R34 (i.e. 0.63 mg O₂/g biomass/min vs. 1.43 mg O₂/g biomass/min for R34 and YV1 respectively at a dilution rate of 1.2 day⁻¹). Monod parameters, maximum growth rate (μ_max) and half saturation coefficient (K_s) were found to be 7.03 day⁻¹ and 691 mg/L for R34 respectively, compared to 5.49 day⁻¹ and 404 mg/L for YV1 respectively. At low dilution rates (<2.5 day⁻¹), when the substrate is present in low concentrations, the growth yield was much higher in YV1 (0.52) than in R34 (0.37). Although substrate utilization rates were similar between R34 and YV1, the latter showed much higher oxygen uptake rate than did R34 at all dilution rates. When the stability of VHb was tested on agar plates containing 40 μg/L of kanamycin and 100 μg/L of ampicillin,vgb gene containing VHb plasmid in YV1 was stable over 82 days. When survivability under oxygen limited conditions was tested, R34 survived only for 11 days whereas YV1 survived over 25 days in liquid media; in agar plate experiments, R34 did not survive more than 40 days whereas more than 75% of YV1 survived over 110 days.     

A novel photobioreactor with transparent rectangular chambers for cultivation of microalgae

An open tank photobioreactor containing transparent rectangular chambers (TRCs) was developed to improve the photosynthetic efficiency of microalgal cultivation. The TRCs, made of transparent acrylic, conducted light deep into the photobioreactor, especially at high cell concentrations. The average irradiance, I_av, was calculated by Lambert–Beer's law, and was used to determine the light conditions in the cultivation system. The photobioreactor provided large areas of illumination that improved the effective utilization of light energy for microalgae growth and created a good artificial environment for a high rate of cell growth, even at low I_av. The biomass concentration of Chlorella sp. reached 3.745 g L⁻¹ on the 13th day, with biomass productivity of 0.340 g L⁻¹ d⁻¹. The total biomass obtained was 56% more than that of similar culture systems without TRCs.     

Palm oil mill effluent (POME) cultured marine microalgae as supplementary diet for rotifer culture

Malaysia is the world’s leading producer of palm oil products that contribute US$ 7.5 billion in export revenues. Like any other agro-based industries, it generates waste that could be utilized as a source of organic nutrients for microalgae culture. Present investigation delves upon Isochrysis sp. culture in POME modified medium and its utilization as a supplement to Nanochloropsis sp. in rotifer cultures. The culture conditions were optimized using a 1 L photobioreactor (Temp: 23°C, illumination: 180 ∼ 200 μmol photons m⁻²s⁻¹, n = 6) and scaled up to 10 L outdoor system (Temp: 26–29°C, illumination: 50 ∼ 180 μmol photons m⁻²s⁻¹, n = 3). Algal growth rate in photobioreactor (μ = 0.0363 h⁻¹) was 55% higher compared to outdoor culture (μ = 0.0163 h⁻¹), but biomass production was 1.3 times higher in outdoor culture (Outdoor = 91.7 mg m⁻²d⁻¹; Photobioreactor = 69 mg m⁻²d⁻¹). Outdoor culture produced 18% higher lipid; while total fatty acids (FA) was not significantly affected by the change in culture systems as both cultures yield almost similar concentrations of fatty acids per gram of sample (photobioreactor = 119.17 mg g⁻¹; outdoor culture = 104.50 mg g⁻¹); however, outdoor cultured Isochrysis sp. had 26% more polyunsaturated fatty acids (PUFAs). Rotifers cultured in Isochrysis sp./ Nanochloropsis sp. (1:1, v/v) mixture gave similar growth rate as 100% Nanochoropsis sp. culture (μ = 0.40 d⁻¹), but had 45% higher counts of rotifers with eggs (t = 7, maximum). The Isochrysis sp. culture successfully lowered the nitrate (46%) and orthophosphate (83%) during outdoor culture.     

Cultivation of Microalgae in Dairy Farm Wastewater Without Sterilization

The present study investigated the feasibility of cultivating microalgae in dairy farm wastewater. The growth of microalgae and the removal rate of the nutrient from the wastewater were examined. The wastewater was diluted 20, 10 and 5 times before applied to cultivate microalgae. A 5 dilution yielded 0.86 g/L dry weight in 6 days with a relative growth rate of 0.28 d^?1, the 10× dilution gave 0.74 g/L and a relative growth rate of 0.26 d^?1 while the 20× dilution 0.59 g/L and a relative growth rate 0.23 d^?1. The nutrients in the wastewater could be removed effectively in different diluted dairy wastewater. The greatest dilution (20×) showed the removal rates: ammonia, 99.26%; P, 89.92%; COD, 84.18%. A 10× dilution removal% was: ammonia 93; P 91 and COD 88. The 5× dilution removal% was: ammonia 83; P 92; COD 90.     

Xanthan batch fermentations: compared performances of a bubble column and a stirred tank fermentor

Fermentations of Xanthomonas campestris, NRRL B-1459, were carried out in a bubble column fermentor (BCF) and in a stirred tank fermentor (STF) to allow comparison of representative variables measured during the microbial growth and the gum production. The microbial growth phase was described by a logistic rate equation where maximum cell concentration was provided by nitrogenous compounds balance. The average value of the maximum specific growth rate was higher in the bubble column (μ _M =0.5 h^?1) than in the stirred reactor (μ _M =0.4 h^?1). The upper values of xanthan yield (Y _g-x =0.65 kg xanthan/kg glucose; Y _{O 2?x} xanthan/kg oxygen) and specific production rate (q _x =0.26 kg xanthan/kg biomass · h) were measured when the oxygen transfer coefficient was kept up above 80 h^?1 in the STF fermentor. In the bubble column the fermentation achieved in the same culture medium lasts two times longer than in the stirred aerated tank; this was attributed to the low value of the oxygen transfer coefficient (K _L a =20 h^?1) at the beginning of the gum synthesis phase. The results obtained in the stirred tank were the basis to estimate the optimal biomass concentration which enables to achieve a culture in non-limiting oxygen transfer conditions. Nevertheless, the transfer characteristics were more homogeneous in the bubble column than in the stirred tank where dead stagnant zones were observed. This is of primary importance when establishing fermentation kinetics models.     

Modeling of growth kinetics for an isolated marine bacterium,Oceanimonas sp. BPMS22 during the production of a trypsin inhibitor

Abstract

Protease inhibitors significantly control physiologically relevant protease activities. Protease inhibitors from marine microbial sources are unique due to their rough living environmental conditions. In the present study, a protein protease inhibitor (PI) was produced from marine Oceanimonas sp. BPMS22. Seven different media were screened for the growth of the bacterium and production of PI. Different carbon and nitrogen sources were screened and optimized for the specific protease inhibitor activity. Three different growth models were checked for the best fit of the bacterial growth. A modified Gompertz model was selected as the best model for the growth of Oceanimonas sp. BPMS22 with the maximum specific growth rate of 0.165?hr^?1 and doubling time of 4.2?hr. The production of PI takes place during the non-growing phase of the bacterial growth. A kinetic model for the production of PI during non-growing phase was used for studying various process parameters. From the model, the maximum trypsin inhibitor formation rate of 0.3802?IU per mg of biomass per hour was observed at 49.91?hr.