Detection and monitoring of viruses are essential for healthy plants and prosperity. Recent development in CRISPR/Cas system in diagnosis has open an avenue well suited for pathogen detection. Variety of CRISPR associated proteins are being discovered, suggesting array of application and detection strategies in diagnosis. Phytopathogenic viruses are diverse with respect to their nucleic acid compositions, which presents a challenge in developing a single device applicable for almost all viruses. The review describes about the efficient use of CRISPR/Cas Technology in diagnosis, such as SHERLOCK, DETECTR and SATORI. These methods are different in their characteristic to identify specific nucleic acids and processing the detectable signals. These technologies are in their infancy and lot of scope is there to develop commercial kits. Plant tissue culture-based industries, climate control green houses, indoor cultivation facilities etc. has been considered as few examples. This review will be beneficial for researchers seeking to develop detection mechanism based on CRISPR/Cas technology. The outcome in the form of cost-effective detection of viruses will be boon for agro-based industries, which are facing challenges through virus contamination.

Biofuels from microalgae have promising potential for a sustainable bioeconomy. Algal strains' oil content and biomass yield are the most influential cost drivers in the fourth generation biofuel (FGB) production. Genetic modification is the key to improving oil accumulation and biomass yield, consequently developing the bioeconomy. This paper discusses current practices, new insights, and emerging trends in genetic modification and their bioeconomic impact on FGB production. It was demonstrated that enhancing the oil and biomass yield could significantly improve the probability of economic success and the net present value of the FGB production process. The techno-economic and socioeconomic burden of using genetically modified (GM) strains and the preventive control strategies on the bioeconomy of FGB production is reviewed. It is shown that the fully lined open raceway pond could cost up to 25% more than unlined ponds. The cost of a plastic hoop air-supported greenhouse covering cultivation ponds is estimated to be US 60,000$ /ha. The competitiveness and profitability of large-scale cultivation of GM biomass are significantly locked to techno-economic and socioeconomic drivers. Nonetheless, it necessitates further research and careful long-term follow-up studies to understand the mechanism that affects these parameters the most.

As the world's population grows, it is necessary to rethink how countries throughout the world produce food in order to replace the conventional and unsustainable agricultural techniques. Microalgae cultivation using a nutrient-rich solution from hydroponic systems not only presents a novel approach to solving problems pertaining to the impact of the discharges on the natural environment but also provides a plethora of other biotechnological applications particularly in the productions of high value-added products and plants growth stimulants, which can be potentially assimilated into the circular bioeconomy (CBE) in the hydroponic sector. In this review, the potential and practicability of microalgae to be merged into hydroponics CBE are reviewed. Overall, the integration of microalgal biorefineries in hydroponics systems can be realized after considering their Technology Readiness Level and System Readiness Level beforehand. Several suggestions on strains and hydroponics system improvement using existing biotechnological tools, Artificial Intelligence (AI) and nanobiotechnology in support of the CBE will be covered.

Algae are effective predecessors of nutrient foods and preventive drugs, gaining global attraction in recent years. It exhibits potent antiviral, antibacterial, antifungal, anti-inflammatory, antioxidant, anti-glycemic, and cholesterol-lowering properties due to their richness in highly valuable secondary metabolites. Nevertheless, algae produce valuable bioproducts, its application in dentistry is in its primitive stage. This review focuses on the emergence and emerging role of micro/macroalgae as a natural source of therapeutic, preventive, and biocompatible agents in dentistry. Several studies unveiled that Cyanobacteria, Spirulina, and Chlorella species offer high oral antibacterial and antifungal properties compared to gold standard agents. The characteristic of algae to scavenge superoxide and hydroxyl free radicals, fabricate them as an anti-oxidative and anti-cancer agent. Either alone or by synergism with pinnacle therapies they are found to produce promising curative actions against periodontitis by embattling proinflammatory cytokines. Technologies extend the functions of microalgae as a detoxifying agent, potent drug delivery system, and adjunct regenerative material in chronic periodontitis. Its application as thickening, binding, anticariogenic agent in toothpaste, antibacterial agent in mouthwash, and biocompatible agent in dental impression materials remains very primitive. Low-cost and eco-friendly technologies are needed for the production of oral hygiene products using algal biomass.

Decomposition of lignocellulosic plant biomass by four filamentous fungi was carried out to facilitate subsequent anaerobic degradation and biogas formation. Agricultural side products, wheat straw and corn stover and forestry energy plant willow chips were selected as plant biomass sources. The substrates were confronted by pure cultures of Penicillium aurantiogriseum (new isolate from rumen), Trichoderma reesei (DSM768), Gilbertella persicaria (SZMC11086) and Rhizomucor miehei (SZMC11005). In addition to total cellulolytic filter paper degradation activity, the production of endoglucanase, cellobiohydrolase, β-glucosidase enzymes were followed during the pretreatment period, which lasted for 10 days at 37 °C. The products of pretreatments were subsequently tested for mesophilic biogas production in batch reactors. All 4 strains effectively pretreated the lignocellulosic substrates albeit in varying degrees, which was related to the level of the tested hydrolytic enzyme activities. Penicillium aurantiogriseum showed outstanding hydrolytic enzyme production and highest biogas yield from the partially degraded substrates. Corn stover was the best substrate for biomass decomposition and biogas production. Scanning electron microscopy confirmed the deep penetration of fungal hyphae into the lignocellulosic substrate in all cases.

An emerging biofilm immobilization method has enabled effortless biomass harvesting and promoted economic feasibility. The current limitation towards the adaptation of this technology is the inadequate understanding of the biofilm interaction towards microporous membrane. Cell adhesion is recognized as the most important step towards the immobilized cultivation of microalgae. Cell attachment kinetic was studied in a short-term batch culture of three marine diatoms, Amphora coffeaeformis, Cylindrotheca fusiformis and Navicula incerta over 96 h on submerged commercial polyvinylidene fluoride (PVDF) membrane under swirling motion of culture medium. Both the evolution of cell adhesion intensity and compositional changes of the extracellular polymeric substances (EPS) released were quantified throughout the cultivation period. To delve into the cell-substratum interactions, existing thermodynamics and colloidal extended Derjaguin, Landau, Vervey, and Overbeek (XDLVO) theory were employed. As a result, A. coffeaeformis and N. incerta recorded a higher cell colonization percentage than C. fusiformis being the lowest about 2.16±0.17% cell colonization due to their respective species-dependent EPS variation. Polysaccharide contents were at least two times higher than protein contents for both C. fusiformis and N. incerta except for A. coffeaeformis depicting a lower polysaccharide-to-protein ratio whereby the protein contents were maximized at 1.03 × 103 ± 64.14 pg m-2 cell-1 at 6th h. From the surface free energy point of view, both thermodynamics and XDLVO model elucidated that cells adhered reversibly in the secondary energy minimum and ranked C. fusiformis the lowest adhesion tendency among three. These findings establish fundamental knowledge about biofilm formation in porous substrate bioreactors.

Cyanobacteria are noteworthy hosts for industrially relevant redox reactions, owing to a light-driven cofactor recycling system using water as electron donor. Customizing Synechocystis sp. PCC 6803 chassis by redirecting electron flow offers a particularly interesting approach to further improve light-driven biotransformations. Therefore, different chassis expressing the heterologous ene-reductase YqjM (namely ΔhoxYH, Δflv3, ΔndhD2 and ΔhoxYHΔflv3) were generated/evaluated. The results showed the robustness of the chassis, that exhibited growth and oxygen evolution rates similar to Synechocystis wild-type, even when expressing YqjM. By engineering the electron flow, the YqjM light-driven stereoselective reduction of 2-methylmaleimide to 2-methylsuccinimide was significantly enhanced in all chassis. In the best performing chassis (ΔhoxYH, lacking an active bidirectional hydrogenase) a 39 % increase was observed, reaching an in vivo specific activity of 116 U gDCW-1 and an initial reaction rate of 16.7 mM h-1. In addition, the presence of the heterologous YqjM mitigated substrate toxicity, and the conversion of 2-methylmaleimide increased oxygen evolution rates, in particular at higher light intensity. In conclusion, this work demonstrates that rational engineering of electron transfer pathways is a valid strategy to increase in vivo specific activities and initial reaction rates in cyanobacterial chassis harboring oxidoreductases.

In the recent two decades, there has been a tremendous increase in the biosynthesis of nanomaterials employing live organisms, their components, extracts, or biomolecules as catalysts. Algae has been used majorly for commercial and industrial uses such as food, feed, skin care, medicines, and fertilizers, algae are now being explored to synthesize green nanoparticles (NPs). Indeed, algae are a rich source of bioactive substances, are easy to produce, grow quickly, and are scalable, therefore this trend is growing by the day. The natural material from algae works as a capping and stabilizing factor in the conversion of metal compounds to metal, metal oxides, or bimetallic NPs. The NPs generated by algae might be intracellular or extracellular, depending on the area of the NPs. The aim of the present review, the first of its kind, is to provide readers with essential information about the diversity of algal strains exploited in the booming field of nanobiotechnology and to explore the biomedical applications of NPs biosynthesized from algae which include antimicrobial, antioxidant, anticancer and biocompatibility properties. Furthermore, this study examines the rationale for the algal-mediated creation of metal, metal oxide, and bimetallic NPs from a variety of algae, as well as the characterization of algae-mediated nanomaterial synthesis.

Biological wastewater treatment is a promising and environmentally friendly method that utilises living microorganisms to remediate water and enable recovery or conversion of contaminants into valuable products. For many decades, microalgae and cyanobacteria, photosynthetic living microorganisms, have been explored extensively for wastewater bioremediation. They can be used for recovering valuable nutrients such as nitrogen and phosphorous from secondary effluents and capable of transforming those nutrients into marketable products such as biofuels, biofertilisers, nutraceutical, and pigments for promoting a Bio-Circular Green economy. In recent years, there has been a shift towards mixing compatible microalgae with bacteria, which is inspired by their natural symbiotic relationships to increase nitrogen and phosphorus recoveries. With this enhanced bioremediation, recovery of polluted wastes can be intensified and higher biomass quality (with high nutrient density) can be achieved. This review focuses on the state-of-the-art of mixed microalgal-bacterial cultivating systems. A comprehensive comparison of existing studies that used Chlorella species as microalgae in various mixed microalgal-bacterial cultivating systems (suspension, biofilm, and immobilisation) for nitrogen and phosphorus recoveries from wastewater is conducted. Key technical challenges such as balancing microalgae and bacteria species, pH regulation, light distribution, biomass harvesting, and biomass conversion are also discussed. From the data comparisons among different cultivation systems, it has been suggested that immobilisation appears to require less amount of operational light compared to the suspended and biofilm-based systems for similar nitrogen and phosphorus removal efficiencies.

Bioconjugates, such as antibody-drug conjugates or fluorescent-labeled proteins, are highly interesting for various applications in medicine and biology. In their production, not only the synthesis is challenging but also the downstream processing, for which hydrophobic interaction chromatography (HIC) is often used. However, in-depth studies of the adsorption of bioconjugates in HIC are still rare. Therefore, in the present work, three different conjugates of lysozyme and fluorescein isothiocyanate (FITC) were synthesized and isolated, and their adsorption on the hydrophobic resin Toyopearl PPG-600 M was systematically studied in batch experiments. The influence of sodium chloride and ammonium sulfate with ionic strengths up to 2000 mM on the adsorption isotherms was investigated at pH 7.0 and 25 °C, and the results were compared to those for pure lysozyme. The conjugation leads to an increase of the adsorption in all studied cases. All studied conjugates contain only a single FITC and differ only in the position of the conjugation on the lysozyme. Despite this, strong differences in the adsorption behavior were observed. Moreover, a mathematical model was developed, which enables the prediction of the adsorption isotherms in the studied systems for varying ionic strengths.

Polyhydroxyalkanoates (PHA) represent an environmentally friendly alternative to petroleum based plastics for a broad range of applications from packaging to biomedical devices. In the prospect of an industrial PHA production, it is highly valuable to accurately control the incorporation of different repeating units into the polymer, to produce a polyester with specific material characteristics. In this study, we develop macroscopic dynamic models predicting the polymer production and composition when mixtures containing up to four volatile fatty acids (VFA) are used as substrates. These models successfully reproduce the sequential (and preferential) substrate consumption and polymer production/reconsumption patterns, experimentally observed during biomass growth, thanks to simple kinetic structures based on Monod and inhibition factors. These models can serve as a basis for numerical simulation and process analysis, as well as process intensification through model-based optimization and control.

The economic production of bioethanol as a sustainable liquid fuel is particularly needed and attractive. Giant reed as a low-cost and renewable biomass can be utilized as a sustainable feedstock for bioethanol development. The current research focuses on optimizing the fermentation parameters to increase ethanol concentration while lowering production costs. In this work, the giant reed was hydrolyzed thermochemically using HCl; cellulose and hemicellulose fractions were maximally converted at optimized hydrolysis conditions (5% HCl, 30 min, and 120 °C), resulting in a high sugar concentration (≈ 55 g/L), which were fermented by Candida tropicalis Y-26 for bioethanol production (≈ 15 g/L). Taguchi design was used to optimize the fermentation parameters (temperatures, pH, incubation period, and nitrogen sources). Under optimum fermentation conditions (25 °C; 24 h.; pH 5.5; and ammonium nitrate as a nitrogen source), the ethanol concentration at flask level accomplished ≈ 21 g/L, while its scale-up to bioreactor level contributed ≈ 25 g/L (equivalent to 250 kg ethanol/ton biomass) with ≈ 67% increase than the fermentation under unoptimized conditions. Overall, these findings proved that optimizing the fermentation parameters by Taguchi design and scaling up at a bioreactor could improve bioethanol production from giant reed biomass.

Fermentation technology is commonly used as a mature process to produce numerous products with the help of micro-organisms. However, these organisms are sometimes inhibited by the accumulation of these products or their by-products. One route to circumvent this is via extractive fermentation, which combines the fermentation process with extraction. To facilitate this, novel bioreactor designs are required, such as the semi-partition bioreactor (SPB) which has been recently proposed for in-situ extractive fermentation. The latter combines a fermentation and an extraction unit into a single vessel using a mixer-settler principle. Where the bioproduct is produced in the mixer and removed continuous in the settler. As the SPB functionality is a subject of interest, this study builds on demonstrating different process conditions in the production of a sample bioprocess (lactic acid (LA)) which is susceptible to product inhibition. The results showed a 34.5 g/L LA concentration was obtained in the pH-controlled condition. While LA production can suffer from product inhibition, neutralizing agents can be easily used to curb inhibitory problems, however, the LA fermentation is a simple (and well-studied) example, which can demonstrate an alternative route to avoiding product inhibition (for systems which cannot be rescued using pH control). Hence, to replicate a scenario of product inhibition, two different process conditions were investigated, no pH control with no extraction (non-integrated), and no pH control with integrated extractive fermentation. Key findings showed higher LA concentration in integrated (25.10 g/L) as compared to the non-integrated (14.94 g/L) case with improved yield (0.75 gg-1 (integrated) versus 0.60 gg-1 (non-integrated)) and overall productivity (0.35 gL-1h-1(integrated) versus 0.20 gL-1h-1(non-integrated)) likewise. This is the first demonstration of an SP bioreactor, and shows how the reactor can be applied to improve productivity. Based on these results, the SPB design can be applied to produce any product liable to product inhibition.

As a part of the search for environment-friendly biocontrol of mosquito-borne diseases, mosquito larvicidal potential of Bacillus thuringiensis subsp. jegathesan (Btj) Cry toxins is explored for toxins with increased toxicity. Safe delivery of the Cry toxins to mosquito larvae in aquatic habitats is a major concern. This is because in water bodies Bacillus thuringiensis (Bt) protein formulations degrade by sunlight, can sink down and get adsorbed by the silt. So, because of its short persistence the toxin requires repeated applications at the given site. Therefore, an upcoming approach is incorporating the Bt toxins in Chlamydomonas reinhardtii (C. reinhardtii) because it is a food of mosquito larvae in water and its molecular toolkit is well investigated for foreign gene expression. The present work aimed to compare the feasibility of C. reinhardtii chloroplast and nuclear compartments for stable expression of Cry11Ba toxin as this is the most toxic Btj protein to date, lethal to different mosquito species. With chloroplast expression of cry11Ba gene we were able to generate marker-free C. reinhardtii strain stably expressing Cry11Ba protein and demonstrating mortality against Aedes aegypti larvae. Moreover, for nuclear expression linking the cry11Ba gene to zeocin via foot and mouth disease virus (FMDV) 2A peptide resulted in the selection of transformants with increased cry11Ba mRNA expression levels by semi-quantitative reverse transcriptase PCR. Obtained results lay a foundation for the C. reinhardtii chloroplast expression system to be used for genetic engineering with Bt toxins which possess enhanced toxicity.

The thermal stability of enzymes dramatically limits their application in the industrial field. Based on the crystal structure, we conducted a semi-rational design according to the B-factor and free energy values to improve the stability of dextranase from Streptococcus mutans (SmdexTM). The B-factor values of Asn102, Asn503, Asp501 and Asp500 were the highest predicted by B-FITTER. Then Rosetta was used to simulate the saturation mutations of Asn102, Asn503, Asp501 and Asp500. The mutated amino acid was designed according to the change of acG. The results showed that the thermal stability of N102P, N102C, D500G, and D500T was improved, and the half-lives of N102P/D500G and N102P/D500T at 45 °C were increased to 3.14 times and 2.44 times, respectively. Analyzing the interaction of amino acids by using Discovery Studio 4.5, it was observed that the thermal stability of dextranase was improved due to the increase in hydrophobicity and the number of hydrogen bonds of the mutant enzyme. The catalytic efficiency of N102P/D500T was increased. Compared with the hydrolyzed products of SmdexTM, the mutant enzymes do not change the specificity of hydrolysates.

Lactobacillus rhamnosus GG (LGG) is one of the most widely used probiotics because of its health benefits and safety. Fucose is among the most abundant hexoses in the human intestine, and LGG consumes fucose to produce energy or proliferate. However, no study has elucidated the metabolism by which LGG metabolizes fucose to produce energy, biomass, and extracellular metabolites. We used metabolomics and flux balance analysis to elucidate these mechanisms and highlight how they might affect the host. We found three different metabolic flux modes by which LGG anaerobically metabolizes fucose to produce energy and biomass. These metabolic flux modes differ from homolactic or heterolactic fermentation and account for the production of lactic acid, 1,2-propanediol, acetic acid, formic acid, and carbon dioxide as a result of fucose metabolism in LGG. We also used gas chromatography/time-of-flight mass spectrometry to identify a variety of short-chain fatty acids and organic acids secreted during fucose metabolism by LGG. Our study is the first to elucidate the unique fucose metabolism of LGG in anaerobic condition.

Nannochloropsis gaditana is a promising microalga for biotechnology. One of the strategies to stimulate its full potential in metabolite production is exposure to flashing lights. Here, we report how N. gaditana adapts to different flashing light regimes (5, 50, and 500 Hz) by changing its cellular physiology and the relative expression of genes related to critical cellular functions. We analyzed the differential mRNA abundance of genes related to photosynthesis, nitrogen assimilation and biosynthesis of chlorophyll, carotenoids, lipids, fatty acids and starch. Analysis of photosynthetic efficiency and high mRNA abundance of photoprotection genes supported the inference that excess excitation energy provided by light absorbance during photosynthesis was produced under low frequency flashing lights and was dissipated by photopigments via the xanthophyll-cycle. Increased relative expression levels of genes related to the synthesis of carotenoids and chlorophyll confirmed the accumulation of photopigments previously observed at low frequency flashing lights. Higher differential mRNA abundance of genes related to the triacylglycerol biosynthesis were observed at lower frequency flashing lights, possibly triggered by a poor nitrogen assimilation caused by low mRNA abundance of a nitrate reductase gene. This study advances a new understanding of algal physiology and metabolism leading to improved cellular performance and metabolite production.

Ionic liquids (ILs) are widely applied in many bioprocesses involving microorganisms due to their unique properties. In this work, the toxicity of imidazolium and pyrrolidinium ionic liquids towards E. coli., a bacterium for which there are limited toxicity data in the literature, was determined. For its simplicity, the nephelometry method was used to estimate ionic liquid toxicity values. The influence of the cation and the alkyl chain length of the cation and anion was analysed. Pyrrolidinium cations were seen to be less toxic than imidazolium cations, while an increase in the alkyl chain length of both pyrrolidinium and imidazolium cations increased the toxicity. Among the anions studied, dimethylphosphate ([Me2PO4]) was the less toxic, while the EC50 for the ionic liquid 1-butyl-3-methylpyrrolidinium dimethylphosphate ([C1C4Pyr][Me2PO4]) was close to 200 mM. Furthermore, a dicationic ionic liquid based on imidazolium and pyrrolidinium cations was synthetized and its toxicity toward E. coli was analysed, maintaining a growth rate of 100 % in the range 0-0.76 mM. The methodology used in this work allows to easily find the less toxic ionic liquids that are biocompatible with E. coli to be used in new bioprocesses.

This study has employed mammalian transient expression systems to generate afucosylated antibodies and antibody Fc mutants for rapid candidate screening in discovery and early development. While chemical treatment with the fucose analogue 2-fluoro-peracetyl-fucose during transient expression only partially produced antibodies with afucosylated N-glycans, the genetic inactivation of the FUT8 gene in ExpiCHO-S™ by CRISPR/Cas9 enabled the transient production of fully afucosylated antibodies. Human IgG1 and murine IgG2a generated by the ExpiCHOfut8KO cell line possessed a 8-to-11-fold enhanced FcγRIIIa binding activity in comparison with those produced by ExpiCHO-S™. The Fc mutant S239D/S298A/I332E produced by ExpiCHO-S™ had an approximate 2-fold higher FcγRIIIa affinity than that of the afucosylated wildtype molecule, although it displayed significantly lower thermal-stability. When the Fc mutant was produced in the ExpiCHOfut8KO cell line, the resulting afucosylated Fc mutant antibody had an additional approximate 6-fold increase in FcγRIIIa binding affinity. This synergistic effect between afucosylation and the Fc mutations was further verified by a natural killer (NK) cell activation assay. Together, these results have not only established an efficient large-scale transient CHO system for rapid production of afucosylated antibodies, but also confirmed a cooperative impact between afucosylation and Fc mutations on FcγRIIIa binding and NK cell activation.

There have been growing interests in microalgal biotechnology for the biorefining of bioactive compounds such as carotenoid pigments, ω-3 fatty acids, antioxidants or antimicrobials for sectoral applications in the pharmacology, nutraceutical and cosmetic fields. This study focused on the unicellular marine rhodophyte Porphyridium purpureum CCAP 1380/1 A, which was cultivated via a two-stage batch growth mode for 10 days using hydrogen peroxide (H2O2), the phytohormone methyl jasmonate (MJ) and three plant extracts (Passiflora incarnata, Panax ginseng and Valeriana officinalis). The microalgal biomass was then analysed for its protein, phycoerythtin, carbohydrate and pigment composition together with its pigment content and antioxidant activity. Of note, MJ increased the protein and phycoerythtin content (up to 225 µg BSA eq./mg DW and 15 mg/ml, respectively) while both the MJ and H2O2 treatments increased carotenoid pigment yields (β-carotene and zeaxanthin, up to 5 and 4 mg/g, respectively). Carbohydrates were enhanced ∼10 fold by the Valeriana officinalis treatment (up 192 μg starch eq./mg). Overall, neutral lipids and antioxidants were mostly negatively affected by the plant extracts. The greatest antioxidant activity registered was obtained with the H2O2 treatment (15 μmol Trolox eq./g DW with TEAC assay). P. purpureum contains multiple valuable compounds of commercial interest. These results indicate that they can be favorably modulated using specific cultivation regimes and chemical enhancers, thereby facilitating the exploitation of the biomass by applying a suitable co-refinery pipeline.

The roots of Pyrenacantha volubilis contain camptothecin (CPT), a high-value bioactive compound possessing anticancer and anti-HIV properties. Isolated root cultures of P. volubilis established in half MS media fortified with 0.3 mgL-1 indole-3-acetic acid and 0.2 mgL-1 indole-3-butyric acid and transferred to light conditions resulted in induction of green roots which obtained a maximum biomass content of 1.09 ± 0.03 g fresh weight with a growth index of 2.07 ± 0.02 in 60 days. The chlorophyll and camptothecin content of the roots was found to increase throughout the culture period attaining a maximum total chlorophyll content of 2.97 ± 0.004 mgg-1 fresh weight in 60 days and CPT content of 2.59 ± 0.02 mgg-1dry weight in 50 days. In the elicitation studies, the green roots treated with 200 mgL-1 yeast extract yielded a maximum camptothecin content of 5.3 ± 0.40 mgg-1 dry weight in 7 days of incubation, which was a 1.94 fold increase in camptothecin content than the control cultures. The thin layer chromatography, high performance liquid chromatography, and liquid chromatography-mass spectroscopy analysis further confirmed the presence of camptothecin (2.17 ± 0.04 mgg-1DW) in the 30 day old in vitro root samples. The overall results suggest the feasibility of green root cultures of P. volubilis as an efficient system for sustainable in vitro production of camptothecin provided further scaling up experiments in bio reactors are imperative.

Microalgae lipid accumulation can be accomplished by different strategies rather than naturally reaching the stationary phase. Many studies employ nitrogen (N) depletion to improve lipid production; however, this approach might not be a suitable alternative when growth in wastewater is attempted. Agro-industry effluents in particular can have high concentrations of N, so nutrient removal is also required. This study evaluated two possibilities of achieving stress conditions in Desmodesmus sp. cultivation: light intensity and CO2 concentration. The culture medium also included liquid and solid residues from the sugarcane agro-industry: vinasse and a biofertilizer produced from bagasse biochar. Optimization of growth in a flat plate photobioreactor was conducted by combining a two-level factorial design and simplex methodology. Both the highest biomass and polyunsaturated fatty acid productivities (150.2 and 21.4 mg L-1 day-1, respectively) were achieved near the central points (5% CO2 in air and 1000 μmol m-2 s-1 light intensity). These results show the possibility of microalgae growth in a sustainable medium coupled with high-value lipid production, e.g., omegas-3, - 6, and - 9.