Application of computational fluid dynamics (CFD) on the raceway design for the cultivation of microalgae: a review.

Microalgae are a potential solution to supersede fossil fuels and produce renewable energy. The major obstacle to the commercialization of microalgae-based biofuels is the high production cost, including nutritional requirements, photobioreactor design, and downstream processes. As for the photobioreactor design, open ponds have been adopted by major commercial plants for their economic advantages. Raceway is a popular type among open ponds. Nevertheless, the fluid dynamics of the raceway operation is quite complex. Software simulation based on Computational Fluid Dynamics is an upcoming strategy for optimizing raceway design. The optimization intends to affect light penetration, particle distribution, mass transfer, and biological kinetics. This review discusses how this strategy can be helpful to design a highly productive raceway pond-based microalgal culture system.

The effects of hydraulic retention time (HRT) on chromium(VI) reduction using autotrophic cultivation of Chlorella vulgaris.

Chromium is an acutely toxic heavy metal that is known to be a carcinogen. Of the two predominant forms of chromium, Cr(III) and Cr(VI), Cr(III) has only about one thousandth the toxicity of Cr(VI). Using microalgal biomass is one way to remove Cr(VI) from the environment. Four days of hydraulic retention time (HRT) was required to completely reduce 10 mg/L of Cr(VI) in the influent. Microalgal biomass is conventionally regarded as an adsorbent in most Cr(VI) reduction studies. However, this study found that Chlorella vulgaris had the potential to convert Cr(VI) to Cr(III) through the enzymatic route of chromium reductase although the measured chromium reductase activity of C. vulgaris was less than that reported values obtained in bacteria. X-ray absorption near-edge spectroscopy (XANES) analysis further showed the absorption edge of Cr(III) in Cr(VI)-treated C. vulgaris, supporting the assumption of Cr(VI) potentially being converted to less-toxic Cr(III).

The growth of oleaginous Rhodotorula glutinis in an airlift bioreactor on crude glycerol through a non-sterile fermentation process.

While the use of oleaginous Rhodotorula glutinis as a feedstock for biodiesel production is an attractive idea, as it can avoid the pollutions associated with over-consumption of fossil fuels. Nevertheless, the related costs, including the energy required for sterilization, remain a barrier to commercialization. This study thus used a low-pH non-sterile medium, instead of a completely sterilized one, to grow R. glutinis in a 5-L airlift bioreactor. The results show that R. glutinis can grow well at a low pH level of 4.0 and without sterilization of the medium, producing a final biomass of 11.7 g/L. Nevertheless, such a low pH will lead to fewer total lipids accumulation, and so a two-stage process of pH control in a non-sterile batch was proposed. Even this two-stage pH operation was also able to produce a similar final biomass of 11.7 g/L. However, the batch with two-stage pH control had a far higher lipid content of 55 ± 4% as compared to that of 21 ± 3% in the batch grown at pH 4.0. This study shows the potential of the proposed non-sterile process with two-stage pH control applied to the growth of R. glutinis to enhance the total lipid accumulation.

The pH effects on the distribution of 1,3-propanediol and 2,3-butanediol produced simultaneously by using an isolated indigenous Klebsiella sp. Ana-WS5.

Several carbon sources were investigated for the production of 1,3-propanediol (PDO) and 2,3-butanediol (BDO) simultaneously, using an isolated indigenous Klebsiella sp. Ana-WS5. The results indicate that glycerol is a suitable carbon source for both BDO and PDO production. Further investigation suggests that adjustment of the pH could alter the metabolic pathway, which affects the ratio of PDO and BDO obtained. The batch with pH controlled at 7.0 had the highest total diol (PDO + BDO) productivity of 0.86 g/L h and the highest PDO/BDO of 7.67, as compared to a batch with pH controlled at 6.0. However, the batch without pH control could achieve a maximum total diol concentration of 48.1 ± 1.6 g/L and the highest yield of 86 % (total diols produced/glycerol consumed). The effects of pH control on the distribution of PDO and BDO concluded in this study could be further applied to the process design for enhancing PDO or BDO production.

A two-stage cultivation process for the growth enhancement of Chlorella vulgaris.

This study proposes a two-stage cultivation process with an autotrophic growth followed by a mixotrophic process. The results indicated that a two-stage cultivation process using a daily dose of 3 g/L of glucose could achieve 7.4 g/L of biomass, which was about a 64 % increase over simple autotrophic cultivation. In the second stage of mixotrophic cultivation, glucose was regarded as a better carbon source for cell growth, than was glycerol. Linoleic acid (C18:2) would be the primary component in the two-stage cultivation as in the autotrophic cultivation. Even carbon source was provided in the second stage of mixotrophic cultivation; lower light intensity limited the mixotrophic growth, which indicated that photosynthesis still plays an important role in the second stage of mixotrophical cultivation. The final biomass was higher after this two-stage cultivation process, which made it suitable for application in the production scale-up of algal biomass.

Integration of microalgae cultivation and anaerobic co-digestion with dairy wastewater to enhance bioenergy and biochemicals production.

A sequential anaerobic digestion and phycoremediation process was employed to recover nutrients and remove pollutants from dairy wastewater (DW), while simultaneously producing biomethane and biochemicals. Anaerobic digestion of 100% DW achieved a methane content and production rate of 53.7% and 0.17 L/L/d, respectively. This was accompanied by the removal of 65.5% chemical oxygen demand (COD), 86% total solid (TS), and 92.8% volatile fatty acids (VFAs). The anaerobic digestate was then used to grow Chlorella sorokiniana SU-1. Using 25% diluted digestate as the medium, SU-1 could reach 4.64 g/L biomass concentration, with total nitrogen (TN), total phosphorus (TP) and COD removal efficiencies of 77.6%, 87.1% and 70.4%, respectively. The obtained microalgal biomass (contained 38.5% carbohydrates, 24.9% proteins, 8.8% lipids) was used to co-digest with DW, resulting in good methane production performance. Co-digestion with 25% (w/v) algal biomass obtained a higher CH4 content (65.2%) and production rate (0.16 L/L/d) than other ratios.

Utilizing microalgal hydrolysate from dairy wastewater-grown Chlorella sorokiniana SU-1 as sustainable feedstock for polyhydroxybutyrate and ß-carotene production by engineered Rhodotorula glutinis #100-29.

The objective of this study was to explore the potential of utilizing Chlorella sorokiniana SU-1 biomass grown on dairy wastewater-amended medium as sustainable feedstock for the biosynthesis of ß-carotene and polyhydroxybutyrate (PHB) by Rhodotorula glutinis #100-29. To break down the rigid cell wall, 100 g/L of microalgal biomass was treated with 3% sulfuric acid, followed by detoxification using 5% activated carbon to remove the hydroxymethylfurfural inhibitor. The detoxified microalgal hydrolysate (DMH) was used for flask-scale fermentation, which yielded a maximum biomass production of 9.22 g/L, with PHB and ß-carotene concentration of 897 mg/L and 93.62 mg/L, respectively. Upon scaling up to a 5-L fermenter, the biomass concentration increased to 11.2 g/L, while the PHB and ß-carotene concentrations rose to 1830 mg/L and 134.2 mg/L. These outcomes indicate that DMH holds promise as sustainable feedstock for the production of PHB and ß-carotene by yeast.

Integrating anaerobic digestion and microalgae cultivation for dairy wastewater treatment and potential biochemicals production from the harvested microalgal biomass.

Utilizing wastewaters as feedstock for microalgal cultivation has the dual benefits of water-saving and low nutrient costs, with simultaneous remediation of pollutants and generation of value-added biochemical products. This study employed two different strategies to treat raw dairy wastewaters with moderate and high chemical oxygen demand (COD) levels. For moderate-COD dairy wastewater, the wastewater was directly utilized as feedstock for algal cultivation, in which the effects of wastewater dilution ratios and algal inoculum sizes were investigated. The results show that the microalga strain used (Chlorella sorokiniana SU-1) was capable of obtaining a high biomass concentration of 3.2 ± 0.1 g/L, accompanied by 86.8 ± 6%, 94.6 ± 3%, and 80.7 ± 1%, removal of COD, total phosphorus (TP) and total nitrogen (TN), respectively. Meanwhile, the obtained microalgal biomass has lipids content of up to 12.0 ± 0.7% at a wastewater dilution ratio of 50% and an inoculum size of 2 g/L. For high-COD dairy wastewater, an integrated process of anaerobic digestion and microalgal phycoremediation was employed, and the effect of inoculum sizes was also studied. The inoculum size of 2 g/L gave highest biomass production of 4.25 ± 0.10 g/L with over 93.0 ± 2.0% removal of COD, TP, and TN. The harvested microalgal biomass has lipids and protein content of 12.5 ± 2.2% and 18.0 ± 2.2%, respectively. The present study demonstrated potential microalgal phycoremediation strategies for the efficient COD removal and nutrients recovery from dairy wastewater of different COD levels with simultaneous production of microalgal biomass which contains valuable components, such as protein and lipids.

Microalgae as sustainable food and feed sources for animals and humans - Biotechnological and environmental aspects.

Offering a potential solution for global food security and mitigating environmental issues caused by the expansion of land-based food production, the carbon-hunger and nutrient-rich microalgae emerged as a sustainable food source for both humans and animals. Other than as an alternative source for protein, microalgae offer its most valuable nutrients, omega-3 and 6 long-chain polyunsaturated fatty acids where the content can compete with that of marine fish with lower chemicals contamination and higher purity. Furthermore, the colorful pigments of microalgae can act as antioxidants together with many other health-improving properties as well as a natural colorant. In addition, the supplementation of algae as animal feed provides plentiful benefits, such as improved growth and body weight, reduced feed intake, enhanced immune response and durability towards illness, antibacterial and antiviral action as well as enrichment of livestock products with bioactive compounds. The significant breakthrough in algal biotechnology has made algae a powerful "cell factory" for food production and lead to the rapid growth of the algal bioeconomy in the food and feed industry. The first overview of this review was to present the general of microalgae and its potential capability. Subsequently, the nutritional compositions of microalgae were discussed together with its applications in human foods and animal feeds, followed by the exploration of their economic feasibility and sustainability as well as market trends. Lastly, both challenges and future perspectives were also discussed.

Microbial cell factories for the production of polyhydroxyalkanoates.

Pollution caused by persistent petro-plastics is the most pressing problem currently, with 8 million tons of plastic waste dumped annually in the oceans. Plastic waste management is not systematized in many countries, because it is laborious and expensive with secondary pollution hazards. Bioplastics, synthesized by microorganisms, are viable alternatives to petrochemical-based thermoplastics due to their biodegradable nature. Polyhydroxyalkanoates (PHAs) are a structurally and functionally diverse group of storage polymers synthesized by many microorganisms, including bacteria and Archaea. Some of the most important PHA accumulating bacteria include Cupriavidus necator, Burkholderia sacchari, Pseudomonas sp., Bacillus sp., recombinant Escherichia coli, and certain halophilic extremophiles. PHAs are synthesized by specialized PHA polymerases with assorted monomers derived from the cellular metabolite pool. In the natural cycle of cellular growth, PHAs are depolymerized by the native host for carbon and energy. The presence of these microbial PHA depolymerases in natural niches is responsible for the degradation of bioplastics. Polyhydroxybutyrate (PHB) is the most common PHA with desirable thermoplastic-like properties. PHAs have widespread applications in various industries including biomedicine, fine chemicals production, drug delivery, packaging, and agriculture. This review provides the updated knowledge on the metabolic pathways for PHAs synthesis in bacteria, and the major microbial hosts for PHAs production. Yeasts are presented as a potential candidate for industrial PHAs production, with their high amenability to genetic engineering and the availability of industrial-scale technology. The major bottlenecks in the commercialization of PHAs as an alternative for plastics and future perspectives are also critically discussed.

Lactic acid production directly from starch in a starch-controlled fed-batch operation using Lactobacillus amylophilus.

Based on the batch results, we constructed a simplified simultaneous saccharification and fermentation (SSF) model for the simulation of lactic acid production directly from unhydrolyzed potato starch using Lactobacillus amylophilus. The results of batch operation at different initial starch concentrations (20, 40 and 60 g/l) indicated that a higher initial starch concentration would lead to a slightly lower productivity, but would largely decrease the yield. Among that, the batch with 20 g/l of initial starch had the maximum productivity and the maximum yield, which would be 0.31 g/(l h) and 98% (g/g), respectively. In view of increasing the productivity and the final lactic acid concentration, a starch-controlled fed-batch operation with 20 g/l of initial starch was performed. It showed the fed-batch operation with starch controlled at 8 ± 1 g/l by adjusting the starch-feeding rate led to the maximum productivity of 0.75 g/(l h) and the yield of 69%.

Application of computational fluid dynamics to raceways combining paddlewheel and CO2 spargers to enhance microalgae growth.

The present study investigated the effect of light intensity and mixing on microalgae growth in a raceway by comparing the performance of a paddlewheel to a combination of paddlewheel and CO2 spargers in a 20 L raceway. The increase of light intensity was known to be able to increase the microalgal growth rate. Increasing paddlewheel rotation speed from 13 to 30 rpm enhanced C. vulgaris growth by enhancing culture mixing. Simulation results using computational fluid dynamics (CFD) indicated that both the turnaround areas of the raceway and the area opposite the paddlewheel experienced very low flow velocities (dead zones) of less than 0.1 m/min, which could cause cell settling and slow down growth. The simulated CFD velocity distribution in the raceway was validated by actual velocity measurements. The installation of CO2 spargers in the dead zones greatly increased flow velocity. The increase of paddlewheel rotation speed reduced the dead zones and hence increased algal biomass production. By complementing the raceway paddlewheel with spargers providing CO2 at 30 mL/min, we achieved a dry cell weight of 5.2 ± 0.2 g/L, which was about 2.6 times that obtained without CO2 sparging.

Coenzyme Q10 production by Rhodobacter sphaeroides in stirred tank and in airlift bioreactor.

A higher Coenzyme Q(10) (CoQ(10)) concentration of 25.04 mg/l was found in airlift bioreactor than the value of 18.11 mg/l obtained in stirred tank under the aerobic-dark cultivation of Rhodobacter sphaeroides. Aeration rate didn't show obvious impact to CoQ(10) production in airlift bioreactor. The fed-batch operation in airlift bioreactor could increase the biomass concentration and led to the maximum CoQ(10) concentration of 33.91 mg/l measured, but a lower CoQ(10) cell content (3.5 mg CoQ(10)/DCW) was observed in the fed-batch operation as compared to the batch operation. To enhance the CoQ(10) content, an aeration-change strategy was proposed in the fed-batch operation of airlift bioreactor. This strategy led to the maximum CoQ(10) concentration of 45.65 mg/l, a 35% increase as compared to the simple fed-batch operation. The results of this study suggested that a fed-batch operation in airlift bioreactor accompanying aeration-change could be suitable for CoQ(10) production.

Current advances in biological swine wastewater treatment using microalgae-based processes.

There is an exponential increase in swine farms around the world to meet the increasing demand for proteins, resulting in a significant amount of swine/piggery wastewater. The wastewater produced in swine farms are rich in ammonia with high eutrophication potential and negative environmental impacts. Safe methods for treatment and disposal of swine wastewater have attracted increased research attention in the recent decades. Conventional wastewater treatment methods are limited by the high ammonia content and chemical/biological oxygen demand of swine wastewater. Recently, microalgal cultivation is being proposed for the phytoremediation of swine wastewater. Microalgae are tolerant to high ammonia levels seen in swine wastewater and they also ensure phosphorus removal simultaneously. This review first gives a brief overview on the conventional methods used for swine wastewater treatment. Microalgae-based processes for the clean-up of swine wastewater are discussed in detail, with their potential advantages and limitations. Future research perspectives are also presented.

Application of thermo-separating aqueous two-phase system in extractive bioconversion of polyhydroxyalkanoates by Cupriavidus necator H16.

Polyhydroxyalkanoates (PHAs), a family of biodegradable and renewable biopolymers show a huge potential as an alternative to conventional plastics. Extractive bioconversion (in situ product recovery) is a technique that integrates upstream fermentation and downstream purification. In this study, extractive bioconversion of PHAs from Cupriavidus necator H16 was performed via a thermo-separating aqueous two-phase system to reduce the cost and environmental impacts of PHAs production. Key operating parameters, such as polymer concentration, temperature, and pH, were optimized. The strategy achieved a yield and PF of 97.6% and 1.36-fold, respectively at 5% EOPO 3900 concentration, 30 °C fermentation temperature and pH 6. The PHAs production process was also successfully scaled up in a 2 L bioreactor. To the best of our knowledge, this is the first report on extractive fermentation of PHAs from Cupriavidus necator utilizing a thermo-separation system to achieve a better productivity and purity of the target product.

Anaerobic co-digestion of algal sludge and waste paper to produce methane.

The unbalanced nutrients of algal sludge (low C/N ratio) were regarded as an important limitation factor to anaerobic digestion process. Adding high carbon content of waste paper in algal sludge feedstock to have a balanced C/N ratio was undertaken in this study. The results showed adding 50% (based on volatile solid) of waste paper in algal sludge feedstock increased the methane production rate to 1170+/-75 ml/l day, as compared to 573+/-28 ml/l day of algal sludge digestion alone, both operated at 4 g VS/l day, 35 degrees C and 10 days HRT. The maximum methane production rate of 1607+/-17 ml/l day was observed at a combined 5 g VS/l day loading rate with 60% (VS based) of paper adding in algal sludge feedstock. Results suggested an optimum C/N ratio for co-digestion of algal sludge and waste paper was in the range of 20-25/1.

Current advances on fermentative biobutanol production using third generation feedstock.

Biobutanol is gaining more attention as a potential alternative to ethanol, and the demand for fermentative biobutanol production has renewed interest. The main challenge faced in biobutanol production is the availability of feedstock. Using conventional agricultural biomass as feedstock is controversial and less efficient, while microalgae, the third generation feedstock, are considered promising feedstock for biobutanol production due to their high growth rate and high carbohydrates content. This review is primarily focused on biobutanol production by using carbohydrate-rich microalgal feedstock. Key technologies and challenges involved in producing butanol from microalgae are discussed in detail and future directions are also presented.

Cultivation of oleaginous Rhodotorula mucilaginosa in airlift bioreactor by using seawater.

The enormous water resource consumption is a concern to the scale-up fermentation process, especially for those cheap fermentation commodities, such as microbial oils as the feedstock for biodiesel production. The direct cultivation of oleaginous Rhodotorula mucilaginosa in a 5-L airlift bioreactor using seawater instead of pure water led to a slightly lower biomass being achieved, at 17.2 compared to 18.1 g/L, respectively. Nevertheless, a higher lipid content of 65 ± 5% was measured in the batch using seawater as compared to the pure water batch. Both the salinity and osmotic pressure decreased as the cultivation time increased in the seawater batch, and these effects may contribute to the high tolerance for salinity. No effects were observed for the seawater on the fatty acid profiles. The major components for both batches using seawater and pure water were C16:0 (palmitic acid), C18:1 (oleic acid) and C18:2 (linoleic acid), which together accounted for over 85% of total lipids. The results of this study indicated that seawater could be a suitable option for scaling up the growth of oleaginous R. mucilaginosa, especially from the perspective of water resource utilization.

Growth of oleaginous Rhodotorula glutinis in an internal-loop airlift bioreactor by using lignocellulosic biomass hydrolysate as the carbon source.

The conversion of abundant lignocellulosic biomass (LCB) to valuable compounds has become a very attractive idea recently. This study successfully used LCB (rice straw) hydrolysate as a carbon source for the cultivation of oleaginous yeast-Rhodotorula glutinis in an airlift bioreactor. The lipid content of 34.3 ± 0.6% was obtained in an airlift batch with 60 g reducing sugars/L of LCB hydrolysate at a 2 vvm aeration rate. While using LCB hydrolysate as the carbon source, oleic acid (C18:1) and linoleic acid (C18:2) were the predominant fatty acids of the microbial lipids. Using LCB hydrolysate in the airlift bioreactor at 2 vvm achieved the highest cell mass growth as compared to the agitation tank. Despite the low lipid content of the batch using LCB hydrolysate, this low cost feedstock has the potential of being adopted for the production of ß-carotene instead of lipid accumulation in the airlift bioreactor for the cultivation of R. glutinis.

The synergistic effects for the co-cultivation of oleaginous yeast-Rhodotorula glutinis and microalgae-Scenedesmus obliquus on the biomass and total lipids accumulation.

In this co-culture of oleaginous yeast-Rhodotorula glutinis and microalgae-Scenedesmus obliquus, microalgae potentially acts as an oxygen generator for the growth of aerobic yeast while the yeast mutually provides CO2 to the microalgae as both carry out the production of lipids. To explore the synergistic effects of co-cultivation on the cells growth and total lipids accumulation, several co-culture process parameters including the carbon source concentration, temperature and dissolved oxygen level would be firstly investigated in the flask trials. The results of co-culture in a 5L photobioreactor revealed that about 40-50% of biomass increased and 60-70% of total lipid increased was observed as compared to the single culture batches. Besides the synergistic effects of gas utilization, the providing of trace elements to each other after the natural cells lysis was believed to be another benefit to the growth of the overall co-culture system.