Immobilized microalgae: principles, processes and its applications in wastewater treatment.

Microalgae have emerged as potential candidates for biomass production and pollutant removal. However, expensive biomass harvesting, insufficient biomass productivity, and low energy intensity limit the large-scale production of microalgae. To break through these bottlenecks, a novel technology of immobilized microalgae culture coupled with wastewater treatment has received increasing attention in recent years. In this review, the characteristics of two immobilized microalgae culture technologies are first presented and then their mechanisms are discussed in terms of biofilm formation theories, including thermodynamic theory, Derjaguin-Landau-Verwei-Overbeek theory (DLVO) and its extended theory (xDLVO), as well as ionic cross-linking mechanisms in the process of microalgae encapsulated in alginate. The main factors (algal strains, carriers, and culture conditions) affecting the growth of microalgae are also discussed. It is also summarized that immobilized microalgae show considerable potential for nitrogen and phosphorus removal, heavy metal removal, pesticide and antibiotic removal in wastewater treatment. The role of bacteria in the cultivation of microalgae by immobilization techniques and their application in wastewater treatment are clarified. This is economically feasible and technically superior. The problems and challenges faced by immobilized microalgae are finally presented.

Morphological growth pattern of Phanerochaete chrysosporium cultivated on different Miscanthus x giganteus biomass fractions.

BACKGROUND: Solid-state fermentation is a fungal culture technique used to produce compounds and products of industrial interest. The growth behaviour of filamentous fungi on solid media is challenging to study due to the intermixity of the substrate and the growing organism. Several strategies are available to measure indirectly the fungal biomass during the fermentation such as following the biochemical production of mycelium-specific components or microscopic observation. The microscopic observation of the development of the mycelium, on lignocellulosic substrate, has not been reported. In this study, we set up an experimental protocol based on microscopy and image processing through which we investigated the growth pattern of Phanerochaete chrysosporium on different Miscanthus x giganteus biomass fractions. RESULTS: Object coalescence, the occupied surface area, and radial expansion of the colony were measured in time. The substrate was sterilized by autoclaving, which could be considered a type of pre-treatment. The fastest growth rate was measured on the unfractionated biomass, followed by the soluble fraction of the biomass, then the residual solid fractions. The growth rate on the different fractions of the substrate was additive, suggesting that both the solid and soluble fractions were used by the fungus. Based on the FTIR analysis, there were differences in composition between the solid and soluble fractions of the substrate, but the main components for growth were always present. We propose using this novel method for measuring the very initial fungal growth by following the variation of the number of objects over time. Once growth is established, the growth can be followed by measurement of the occupied surface by the mycelium. CONCLUSION: Our data showed that the growth was affected from the very beginning by the nature of the substrate. The most extensive colonization of the surface was observed with the unfractionated substrate containing both soluble and solid components. The methodology was practical and may be applied to investigate the growth of other fungi, including the influence of environmental parameters on the fungal growth.

Consolidated bioethanol production from olive mill waste: Wood-decay fungi from central Morocco as promising decomposition and fermentation biocatalysts.

Meknes region is a Moroccan olive-processing area generating high amounts of non-valorized Olive Mill Waste (OMW). Fungi are natural decomposers producing varied enzyme classes and effectively contributing to the carbon cycle. However, structural complexity of biomass and modest performances of wild fungi are major limits for local biorefineries. The objective of current research is to assess the ability of local fungi for bioethanol production from OMW using Consolidated Bioprocessing (CBP). This is done by characterizing lignocellulolytic potential of six wood-decay and compost-inhabiting ascomycetes and selecting potent fermentation biocatalysts. High and diversified activities were expressed by Fusarium solani and Fusarium oxysporum: 9.36 IU. mL-1 and 2.88 IU. mL-1 total cellulase activity, 0.54 IU. mL-1 and 0.57 IU. mL-1 laccase activity, respectively, and 8.43 IU. mL-1 lignin peroxidase activity for the latter. F. oxysporum had maximum bioethanol production and yield of 2.47 g.L-1 and 0.84 g.g-1, respectively, qualifying it as an important bio-agent for single-pot local biorefinery.

Current and novel approaches to downstream processing of microalgae: A review.

Biotechnological application of microalgae cultures at large scale has significant potential in the various fields of biofuels, food and feed, cosmetic, pharmaceutic, environmental remediation and water treatment. Despite this great potential application, industrialisation of microalgae culture and valorisation is still faced with serious remaining challenges in culture scale-up, harvesting and extraction of target molecules. This review presents a general summary of current techniques for harvesting and extraction of biomolecules from microalgae, their relative merits and potential for industrial application. The cell wall composition and its impact on microalgae cell disruption is discussed. Additionally, more recent progress and promising experimental methods and studies are summarised that would allow the reader to further investigate the state of the art. A final survey of energetic assessments of the different techniques is also made. Bead milling and high-pressure homogenisation seem to give clear advantages in terms of target high value compounds extraction from microalgae, with enzyme hydrolysis as a promising emerging technique. Future industrialisation of microalgae for high scale biotechnological processing will require the establishment of universal comparison-standards that would enable easy assessment of one technique against another.

A predictive dynamic yeast model based on component, energy, and electron carrier balances.

The present study describes a novel yeast model for the prediction of yeast fermentation. The proposed model considers the possible metabolic pathways of yeast. For each pathway, the time evolution of components, energy (ATP/ADP), and electron carriers (NAD+ /NADH) are expressed with limitation factors for all quantities consumed by each respective pathway. In this manner, the model can predict the partition of these pathways based on the growth conditions and their evolution over time. Several biological pathways and their stoichiometric coefficients are well known from literature. It is important to note that most of the kinetic parameters have no effect as the actual kinetics are controlled by the balance of limiting factors. The few remaining parameters were adjusted and compared with the literature when the data set was available. The model fits our experimental data from yeast fermentation on glucose in a nonaerated batch system. The predictive ability of the model and its capacity to represent the intensity of each pathway over time facilitate an improved understanding of the interactions between the pathways. The key role of energy (ATP) and electron carrier (NAD+ ) to trigger the different metabolic pathways during yeast growth is highlighted, whereas the involvement of mitochondrial respiration not being associated with the TCA cycle is also shown.

Optimize, Modulate, and Scale-up Resveratrol and Resveratrol Dimers Bioproduction in Vitis labrusca L. Cell Suspension from Flasks to 20 L Bioreactor.

Resveratrol and its oligomers are biologically active compounds. This work brings new insights for the bioproduction of trans-resveratrol with three dimers, pallidol, trans-ε-viniferin, and trans-δ-viniferin, in cell suspension of Vitis labrusca. Conditions of elicitation by methyl jasmonate were optimized for the production of stilbenes using statistical design of experiment. Bio-production of stilbenes was scaled-up to 5 L and in these conditions, trans-resveratrol concentrations reached 237 mg/L, and for pallidol 114 mg/L. The comparison of different elicitation modes (different elicitors, combination with cyclodextrins or adsorbent resin) allowed to reach particularly high concentrations of target molecules: Resveratrol 6.14 g/L, pallidol 0.90 g/L, δ-viniferin 0.54 g/L, and ε-viniferin 0.50 g/L. Scale-up to 20 L-stirring-bioreactor gave similar growth rates to those observed in shake flask culture, with a high production of resveratrol (4.23 g/L) and δ-viniferin (0.76 g/L). This work provides new strategies for the production of stilbenes in plant cell suspension for biological and commercial evaluation.

Process for symbiotic culture of Saccharomyces cerevisiae and Chlorella vulgaris for in situ CO2 mitigation.

Industrial biotechnology relies heavily on fermentation processes that release considerable amounts of CO2. Apart from the fact that this CO2 represents a considerable part of the organic substrate, it has a negative impact on the environment. Microalgae cultures have been suggested as potential means of capturing the CO2 with further applications in high-value compounds production or directly for feed applications. We developed a sustainable process based on a mixed co-dominant culture of Saccharomyces cerevisiae and Chlorella vulgaris where the CO2 production and utilization controlled the microbial ecology of the culture. By mixing yeast and microalga in the same culture, the CO2 is produced in dissolved form and is available to the microalga avoiding degassing and dissolution phenomena. With this process, the CO2 production and utilization rates were balanced and a mutual symbiosis between the yeast and the microalga was set up in the culture. In this study, the reutilization of CO2 and growth of C. vulgaris was demonstrated. The two organism populations were balanced at approximately 20 × 106 cells ml-1 and almost all the CO2 produced by yeast was reutilized by microalga within 168 h of culture. The C. vulgaris inoculum preparation played a key role in establishing co-dominance of the two organisms. Other key factors in establishing symbiosis were the inoculum ratio of the two organisms and the growth medium design. A new method allowed the independent enumeration of each organism in a mixed culture. This study could provide a basis for the development of green processes of low environmental impact.

Magnesium Uptake by the Green Microalga Chlorella vulgaris in Batch Cultures.

The accumulation (internal and superficial distribution) of magnesium ions (Mg(2+)) by the green freshwater microalga Chlorella vulgaris (C. vulgaris) was investigated under autotrophic culture in a stirred photobioreactor. The concentrations of the three forms of Mg(2+) (dissolved, extracellular, and intracellular) were determined with atomic absorption spectroscopy during the course of C. vulgaris growth. The proportions of adsorbed (extracellular) and absorbed (intracellular) Mg(2+) were quantified. The concentration of the most important pigment in algal cells, chlorophyll a, increased over time in proportion to the increase in the biomass concentration, indicating a constant chlorophyll/biomass ratio during the linear growth phase. The mean-average rate of Mg(2+) uptake by C. vulgaris grown in a culture medium starting with 16 mg/l of Mg(2+) concentration was measured. A clear relationship between the biomass concentration and the proportion of the Mg(2+) removal from the medium was observed. Of the total Mg(2+) present in the culture medium, 18% was adsorbed on the cell wall and 51% was absorbed by the biomass by the end of the experiment (765 h). Overall, 69% of the initial Mg(2+) were found to be removed from the medium. This study supported the kinetic model based on a reversible first-order reaction for Mg(2+) bioaccumulation in C. vulgaris, which was consistent with the experimental data.

Nitrogen and phosphate removal from wastewater with a mixed microalgae and bacteria culture.

Microalgae are able to convert nutrients (nitrogen and phosphorus) from wastewater into biomass and bio-products, thus improving the sustainability of wastewater treatment. In High Rate Algal Ponds (HRAP), biomass productivity and water treatment efficiency are highly dependent on environmental parameters such as temperature, light intensity and photoperiod. The influence of temperature and photoperiod on biomass productivity and the removal of dissolved nitrogen and phosphorus from municipal wastewater by a native microalgae-bacteria consortium was assessed in batch cultures in view of the development of an HRAP at a larger scale. Temperature affected the growth rate and microalgae biomass production as well as ammonium and phosphate removal rates. At the temperatures 15 and 25 °C, the average total nitrogen and phosphorus removal extents ranged from 72 to 83% and 100% respectively. Additionally 33.0 ± 0.1% of the total nitrogen was eliminated by stripping at 25 °C, and 50 ± 2% was assimilated by the microorganisms under all conditions tested.

Effects of processing steps on the phenolic content and antioxidant activity of beer.

A new analytical method (liquid chromatography-antioxidant, LC-AOx) was used that is intended to separate beer polyphenols and to determine the potential antioxidant activity of these constituents after they were allowed to react online with a buffered solution of the radical cation 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS(•+)). Using the LC-AOx method, it was possible to demonstrate that the extent of the antioxidant activity was very much dependent on the phenolic compound considered. The method was also applied to the analysis of beer extracts and allowed the evaluation of their antioxidant activity at different steps of beer processing: brewing, boiling, and fermentation. This study showed that the total antioxidant activity remained unchanged throughout beer processing, as opposed to the polyphenolic content, which showed a 3-fold increase. Hopping and fermentation steps were the main causes of this increase. However, the increase measured after fermentation was attributed to a better extraction of polyphenols due to the presence of ethanol, rather than to a real increase in their content. Moreover, this method allowed the detection of three unknown antioxidant compounds, which accounted for 64 ± 4% of the total antioxidant activity of beer and were individually more efficient than caffeic acid and epicatechin.