Isolation and evaluation of oil-producing microalgae from subtropical coastal and brackish waters

Microalgae have been widely reported as a promising source of biofuels, mainly based on their high areal productivity of biomass and lipids as triacylglycerides and the possibility for cultivation on non-arable land. The isolation and selection of suitable strains that are robust and display high growth and lipid accumulation rates is an important prerequisite for their successful cultivation as a bioenergy source, a process that can be compared to the initial selection and domestication of agricultural crops. We developed standard protocols for the isolation and cultivation for a range of marine and brackish microalgae. By comparing growth rates and lipid productivity, we assessed the potential of subtropical coastal and brackish microalgae for the production of biodiesel and other oil-based bioproducts. This study identified Nannochloropsis sp., Dunaniella salina and new isolates of Chlorella sp. and Tetraselmis sp. as suitable candidates for a multiple-product algae crop. We conclude that subtropical coastal microalgae display a variety of fatty acid profiles that offer a wide scope for several oil-based bioproducts, including biodiesel and omega-3 fatty acids. A biorefinery approach for microalgae would make economical production more feasible but challenges remain for efficient harvesting and extraction processes for some species.     

The dilemma for lipid productivity in green microalgae: importance of substrate provision in improving oil yield without sacrificing growth

Rising oil prices and concerns over climate change have resulted in more emphasis on research into renewable biofuels from microalgae. Unlike plants, green microalgae have higher biomass productivity, will not compete with food and agriculture, and do not require fertile land for cultivation. However, microalgae biofuels currently suffer from high capital and operating costs due to low yields and costly extraction methods. Microalgae grown under optimal conditions produce large amounts of biomass but with low neutral lipid content, while microalgae grown in nutrient starvation accumulate high levels of neutral lipids but are slow growing. Producing lipids while maintaining high growth rates is vital for biofuel production because high biomass productivity increases yield per harvest volume while high lipid content decreases the cost of extraction per unit product. Therefore, there is a need for metabolic engineering of microalgae to constitutively produce high amounts of lipids without sacrificing growth. Substrate availability is a rate-limiting step in balancing growth and fatty acid (FA) production because both biomass and FA synthesis pathways compete for the same substrates, namely acetyl-CoA and NADPH. In this review, we discuss the efforts made for improving biofuel production in plants and microorganisms, the challenges faced in achieving lipid productivity, and the important role of precursor supply for FA synthesis. The main focus is placed on the enzymes which catalyzed the reactions supplying acetyl-CoA and NADPH.     

Biodiesel production with microalgae as feedstock: from strains to biodiesel

Due to negative environmental influence and limited availability, petroleum-derived fuels need to be replaced by renewable biofuels. Biodiesel has attracted intensive attention as an important biofuel. Microalgae have numerous advantages for biodiesel production over many terrestrial plants. There are a series of consecutive processes for biodiesel production with microalgae as feedstock, including selection of adequate microalgal strains, mass culture, cell harvesting, oil extraction and transesterification. To reduce the overall production cost, technology development and process optimization are necessary. Genetic engineering also plays an important role in manipulating lipid biosynthesis in microalgae. Many approaches, such as sequestering carbon dioxide from industrial plants for the carbon source, using wastewater for the nutrient supply, and maximizing the values of by-products, have shown a potential for cost reduction. This review provides a brief overview of the process of biodiesel production with microalgae as feedstock. The methods associated with this process (e.g. lipid determination, mass culture, oil extraction) are also compared and discussed.     

Tailoring cyanobacterial cell factory for improved industrial properties

Photosynthetic biomanufacturing provides a promising solution for sustainable production of biofuels and biochemicals. Cyanobacteria are among the most promising microbial platforms for the construction of photosynthetic cell factories. Metabolic engineering of cyanobacteria has enabled effective photosynthetic synthesis of diverse natural or non-natural metabolites, while commercialization of photosynthetic biomanufacturing is usually restricted by process and economic feasibilities. In actual outdoor conditions, active cell growth and product synthesis is restricted to narrow light exposure windows of the day-night cycles and is threatened by diverse physical, chemical, and biological environmental stresses. For biomass harvesting and bioproduct recovery, energy and cost consuming processing and equipment is required, which further decreases the economic and environmental competitiveness of the entire process. To facilitate scaled photosynthetic biomanufacturing, lots of efforts have been made to engineer cyanobacterial cell properties required by robust & continual cultivation and convenient & efficient recovery. In this review, we specifically summarized recently reported engineering strategies on optimizing industrial properties of cyanobacterial cells. Through systematically re-editing the metabolism, morphology, mutualism interaction of cyanobacterial chassis cells, the adaptabilities and compatibilities of the cyanobacterial cell factories to the industrial process could be significantly improved. Cell growth and product synthesis of the tailored cyanobacterial cells could be expanded and maintained at night and in stressful environments, while convenient biomass harvesting could also be expected. For developing more feasible cyanobacterial photosynthetic biomanufacturing in large scale, we here propose the importance of tailoring industrial properties of cyanobacteria and outline the directions that should be exploited in the future.     

Cyanobacteria as an eco-friendly resource for biofuel production: A critical review

Cyanobacteria are photosynthetic microorganisms which can be found in various environmental habitats. These photosynthetic bacteria are considered as promising feedstock for the production of the third- and the fourth-generation biofuels. The main subject of this review is highlighting the significant aspects of the biofuel production from cyanobacteria. The most recent investigations about the extraction or separation of the bio-oil from cyanobacteria are also adduced in the present review. Moreover, the genetic engineering of cyanobacteria for improving biofuel production and the impact of bioinformatics studies on the designing better-engineered strains are mentioned. The large-scale biofuel production is challenging, so the economic considerations to provide inexpensive biofuels are also cited. It seems that the future of biofuels is strongly dependent to the following items; understanding the metabolic pathways of the cyanobacterial species, progression in the construction of the engineered cyanobacteria, and inexpensive large-scale cultivation of them.     

Membrane applications for microalgae cultivation and harvesting: a review

With renewed interest in microalgae due to their potential for biofuel and bioproducts production, efficient cultivation and harvesting mechanisms are needed to increase the economic competitiveness of microalgal products against traditional sources. With pore sizes ranging from microns to angstroms, membranes provide tailored functions for solid/liquid separation (cell retention, biomass concentration and dewatering), gas/liquid separation (gas delivery and removal), and solute/liquid separation (bioproduct recovery, feedstock preparation and effluent recycling) that are problematic or not possible with other technologies. Existing knowledge on membrane systems used in other disciplines, such as environmental engineering, marine science, and biomedicine, can be applied to algae production. Though membranes have great potential to facilitate cultivation and harvesting, challenges in energy reduction and fouling mitigation need to be overcome for long-term, cost-effective application.     

Photobioreactor cultivation strategies for microalgae and cyanobacteria

The current burden on fossil‐derived chemicals and fuels combined with the rapidly increasing global population has led to a crucial need to develop renewable and sustainable sources of chemicals and biofuels. Photoautotrophic microorganisms, including cyanobacteria and microalgae, have garnered a great deal of attention for their capability to produce these chemicals from carbon dioxide, mineralized water, and solar energy. While there have been substantial amounts of research directed at scaling‐up production from these microorganisms, several factors have proven difficult to overcome, including high costs associated with cultivation, photobioreactor construction, and artificial lighting. Decreasing these costs will substantially increase the economic feasibility of these production processes. Thus, the purpose of this review is to describe various photobioreactor designs, and then provide an overview on lighting systems, mixing, gas transfer, and the hydrodynamics of bubbles. These factors must be considered when the goal of a production process is economic feasibility. Targets for improving microalgae and cyanobacteria cultivation media, including water reduction strategies will also be described. As fossil fuel reserves continue to be depleted and the world population continues to increase, it is imperative that renewable chemical and biofuel production processes be developed toward becoming economically feasible. Thus, it is essential that future research is directed toward improving these processes. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:811–827, 2018     

Membrane technology in microalgae cultivation and harvesting: A review

Membrane processes have long been applied in different stages of microalgae cultivation and processing. These processes include microfiltration, ultrafiltration, dialysis, forward osmosis, membrane contactors and membrane spargers. They are implemented in many combinations, both as a standalone and as a coupled system (in membrane biomass retention photobioreactors (BR-MPBRs) or membrane carbonation photobioreactors (C-MPBRs). To provide sufficient background on these applications, an overview of membrane materials and membrane processes of interest in microalgae cultivation and processing is provided in this work first. Afterwards, discussion about specific aspects of membrane applications in microbial cultivation and harvesting is provided, including membrane fouling. Many of the membrane processes were shown to be promising options in microalgae cultivation. Yet, significant process optimizations are still required when they are applied to enable microalgae biomass bulk production to become competitive as a raw material for biofuel production. Recent developments of the coupled systems (BR-MPBR and C-MPBR) bring significant promises to improve the volumetric productivity of a cultivation system and the efficiency of inorganic carbon capture, respectively.     

Effects of bacterial communities on biofuel-producing microalgae: stimulation,inhibition and harvesting

Despite the great interest in microalgae as a potential source of biofuel to substitute for fossil fuels, little information is available on the effects of bacterial symbionts in mass algal cultivation systems. The bacterial communities associated with microalgae are a crucial factor in the process of microalgal biomass and lipid production and may stimulate or inhibit growth of biofuel-producing microalgae. In addition, we discuss here the potential use of bacteria to harvest biofuel-producing microalgae. We propose that aggregation of microalgae by bacteria to achieve >90% reductions in volume followed by centrifugation could be an economic approach for harvesting of biofuel-producing microalgae. Our aims in this review are to promote understanding of the effects of bacterial communities on microalgae and draw attention to the importance of this topic in the microalgal biofuel field.     

Cultivation, photobioreactor design and harvesting of microalgae for biodiesel production: A critical review

Microalgae have the ability to mitigate CO₂ emission and produce oil with a high productivity, thereby having the potential for applications in producing the third-generation of biofuels. The key technologies for producing microalgal biofuels include identification of preferable culture conditions for high oil productivity, development of effective and economical microalgae cultivation systems, as well as separation and harvesting of microalgal biomass and oil. This review presents recent advances in microalgal cultivation, photobioreactor design, and harvesting technologies with a focus on microalgal oil (mainly triglycerides) production. The effects of different microalgal metabolisms (i.e., phototrophic, heterotrophic, mixotrophic, and photoheterotrophic growth), cultivation systems (emphasizing the effect of light sources), and biomass harvesting methods (chemical/physical methods) on microalgal biomass and oil production are compared and critically discussed. This review aims to provide useful information to help future development of efficient and commercially viable technology for microalgae-based biodiesel production.     

Strategies to enhance production of microalgal biomass and lipids for biofuel feedstock

ABSTRACT

Microalgae have enormous potential as feedstock for biofuel production compared with other sources, due to their high areal productivity, relatively low environmental impact, and low impact on food security. However, high production costs are the major limitation for commercialization of algal biofuels. Strategies to maximize biomass and lipid production are crucial for improving the economics of using microalgae for biofuels. Selection of suitable algal strains, preferably from indigenous habitats, and further improvement of those ‘platform strains’ using mutagenesis and genetic engineering approaches are desirable. Conventional approaches to improve biomass and lipid productivity of microalgae mainly involve manipulation of nutritional (e.g. nitrogen and phosphorus) and environmental (e.g. temperature, light and salinity) factors. Approaches such as the addition of phytohormones, genetic and metabolic engineering, and co-cultivation of microalgae with yeasts and bacteria are more recent strategies to enhance biomass and lipid productivity of microalgae. Improvement in culture systems and the use of a hybrid system (i.e. a combination of open ponds and photobioreactors) is another strategy to optimize algal biomass and lipid production. In addition, the use of low-cost substrates such as agri-industrial wastewater for the cultivation of microalgae will be a smart strategy to reduce production costs. Such systems not only generate high algal biomass and lipid productivity, but are also useful for bioremediation of wastewater and bioremoval of waste CO₂. The aim of this review is to highlight the advances in the use of various strategies to enhance production of algal biomass and lipids for biofuel feedstock.     

Efficacy of microalgae for industrial wastewater treatment: a review on operating conditions,treatment efficiency and biomass productivity

Sustainable, clean, renewable energy without negotiating contiguous environment is a challenging task mainly comprises of natural resource management which involves operational efficiency, waste minimisation and energy recovery. Disposal of untreated industrial wastewater with chemical nutrients especially compounds containing nitrogen and phosphorous lead to eutrophication and related environmental issues that affect the recycling processes of bio system. Biotransformation of pollutants using microalgae has proven to be proficient and economic method of wastewater treatment due to their adaptability of growing in various wastewater streams and also useful in the process of CO₂ fixation. Moreover this technology has the competence of producing bio fuels as an alternative energy resource in the form of bio diesel, bio ethanol and biogas. In this review paper, the applicability of microalgae cultivation in industrial wastewater treatment has been discussed extensively including the processes involved, influencing operational parameters such as study mode, cultivation mode and time, method of aeration, pH and intensity of light. Further, the cultivation methods, harvesting techniques involved in the treatment process have been presented. In addition, the analysis on removal efficiency of algal treatment, biomass productivity and lipid content of the cultivated biomass has been discussed widely which possibly will be helpful in adopting the process integration in industrial wastewater treatment with bio energy production.     

Harvesting of microalgae by bio-flocculation

The high-energy input for harvesting biomass makes current commercial microalgal biodiesel production economically unfeasible. A novel harvesting method is presented as a cost and energy efficient alternative: the bio-flocculation by using one flocculating microalga to concentrate the non-flocculating microalga of interest. Three flocculating microalgae, tested for harvesting of microalgae from different habitats, improved the sedimentation rate of the accompanying microalga and increased the recovery of biomass. The advantages of this method are that no addition of chemical flocculants is required and that similar cultivation conditions can be used for the flocculating microalgae as for the microalgae of interest that accumulate lipids. This method is as easy and effective as chemical flocculation which is applied at industrial scale, however in contrast it is sustainable and cost-effective as no costs are involved for pre-treatment of the biomass for oil extraction and for pre-treatment of the medium before it can be re-used.     

Mechanism and challenges in commercialisation of algal biofuels

Biofuels made from algal biomass are being considered as the most suitable alternative energy in current global and economical scenario. Microalgae are known to produce and accumulate lipids within their cell mass which is similar to those found in many vegetable oils. The efficient lipid producer algae cell mass has been reported to contain more than 30% of their cell weight as lipids. According to US DOE microalgae have the potential to produce 100 times more oil per acre land than any terrestrial plants. This article reviews up to date literature on the composition of algae, mechanism of oil droplets, triacylglycerol (TAG) production in algal biomass, research and development made in the cultivation of algal biomass, harvesting strategies, and recovery of lipids from algal mass. The economical challenges in the production of biofuels from algal biomass have been discussed in view of the future prospects in the commercialisation of algal fuels.     

Chromatographic capture of cells to achieve single stage clarification in recombinant protein purification

Recent advancements in cell culture engineering have allowed drug manufacturers to achieve higher productivity by driving higher product titers through cell line engineering and high-cell densities. However, these advancements have shifted the burden to clarification and downstream processing where the difficulties now revolve around removing higher levels of process- and product-related impurities. As a result, a lot of research efforts have turned to developing new approaches and technologies or process optimization to still deliver high quality biological products while controlling cost of goods. Here, we explored the impact of a novel single use technology employing chromatographic principle-based clarification for a process-intensified cell line technology. In this study, a 16% economic benefit ($/g) was observed using a single-use chromatographic clarification compared to traditional single-use clarification technology by improving the overall product cost through decreased operational complexity, higher loading capacity, increased product recovery, and higher impurity clearance. In the end, the described novel chromatographic approach significantly simplified and enhanced the cell culture fluid harvest unit operation by combining the reduction of insoluble and key soluble contaminants of the harvest fluid into a single stage.     

Bioflocculation as an innovative harvesting strategy for microalgae

Microalgae are a promising new source of biomass for the production of third generation biofuels but, so far, the majority of microalgal biomass has been used for high-value applications. New low-cost technologies are needed to make the production and processing of microalgae economically feasible for low-value applications. A major challenge lies in the harvesting of microalgae, which requires a cost-efficient separation technology. Flocculation, especially bioflocculation, is an attractive low-cost separation technology. Various new bioflocculation strategies have been claimed to generate major advances in cost-efficient harvesting. Here, we review the recent advances in bioflocculation based on algal–bacterial, algal–fungal, or algal–algal interactions within the framework of microalgae biomass harvesting for biofuel production. We also discuss recent advances using infochemicals and genetic engineering for the induction of bioflocculation.     

Cyanobacteria cultivation in industrial wastewaters and biodiesel production from their biomass: a review

As an alternative fuel biodiesel has become increasingly important due to diminishing petroleum reserves and adverse environmental consequences of exhaust gases from petroleum-fueled engines. Recently, research interest has focused on the production of biofuel from microalgae. Cyanobacteria appeared to be suitable candidates for cultivation in wastes and wastewaters because they produce biomass in satisfactory quantity and can be harvested relatively easily due to their size and structure. In addition, their biomass composition can be manipulated by several environmental and operational factors to produce biomass with concrete characteristics. Herein, we review the culture of cyanobacteria in wastewaters and also the potential resources that can be transformed into biodiesel successfully for meeting the ever-increasing demand for biodiesel production.     

Challenges in the microbial production of flavonoids

As flavonoids have beneficial health effects on humans, they are gaining increasing interest from pharmaceutical and health industries. However, current production methods, such as plant extraction and chemical synthesis, are inadequate to meet the demand. Therefore, microbial production might offer a promising alternative. During recent years, microbial strains able to produce flavonoids to a certain extent have been developed. However production titers are limited to the mg l^?1 range, hampering the industrial exploitation of these strains. The latter will not be achieved by simply introducing the heterologous pathway in the production host and optimizing the fermentation process, but will depend on the interaction of different aspects of metabolic engineering and process engineering to overcome the current limitations. Next to engineering the production strain to optimize the availability of precursors, the pathway itself also requires intensive engineering. Currently utilized strategies result in a wide variety of different production strains, requiring high-throughput screening methods to identify optimal performing strains. As more and more organisms are being characterized, each with their own specific properties which might be beneficial for the heterologous production of flavonoids, the choice of the production host is another important aspect. Finally, the use of co-cultures might offer an alternative in which different parts of the process are performed by different organisms. This review aims to provide an overview of the research that has been done on these separate aspects. The work presented here could be used as a framework for further research.     

Developments and perspectives of photobioreactors for biofuel production

The production of biofuels from microalgae requires efficient photobioreactors in order to meet the tight constraints of energy efficiency and economic profitability. Current cultivation systems are designed for high-value products rather than for mass production of cheap energy carriers. Future bioreactors will imply innovative solutions in terms of energy efficiency, light and gas transfer or attainable biomass concentration to lower the energy demand and cut down production costs. A new generation of highly developed reactor designs demonstrates the enormous potential of photobioreactors. However, a net energy production with microalgae remains challenging. Therefore, it is essential to review all aspects and production steps for optimization potential. This includes a custom process design according to production organism, desired product and production site. Moreover, the potential of microalgae to synthesize valuable products additionally to the energetic use can be integrated into a production concept as well as waste streams for carbon supply or temperature control.