Microalgae protein digestibility: How to crack open the black box?

Microalgae are booming as a sustainable protein source for human nutrition and animal feed. Nevertheless, certain strains were reported to have robust cell walls limiting protein digestibility. There are several disruption approaches to break down the cell integrity and increase digestive enzyme accessibility. This review's intent is to discuss the digestibility of microalgae proteins in intact cells and after their disruption. In intact single cells, the extent of protein digestibility is chiefly related to cell wall structural properties (differing among strains) as well as digestion method and when added to food or feed protein digestibility changes depending on the matrix's composition. The degree of effectiveness of the disruption method varies among studies, and it is complicated to compare them due to variabilities in digestibility models, strains, disruption method/conditions and their consequent impact on the microalgae cell structure. More exhaustive studies are still required to fill knowledge gaps on the structure of microalgal cell walls and to find efficient and cost-effective disruption technologies to increase proteins availability without hindering their quality.

Enhanced short-chain fatty acid production from sludge anaerobic fermentation by combined pretreatment with sodium pyrophosphate and thermal hydrolysis.

The slow breakdown of sludge is the primary obstacle hindering the conversion of waste-activated sludge to short-chain fatty acids (SCFAs) by anaerobic fermentation. This study proposed a novel method incorporating sodium pyrophosphate and thermal hydrolysis (SP-TH) for sludge pretreatment and evaluated its effectiveness regarding SCFA production. The combined pretreatment of SP at 0.4 g/g of total suspended solids and TH at 140 °C enhanced SCFA production from 2,169 ± 208 to 4,388 ± 184 mg chemical oxygen demand/L. SP strips extracellular polymeric substances, and the subsequent TH decomposes cells in the sludge, thus promoting sludge hydrolysis. SP-TH pretreatment promoted SCFA accumulation by enhancing enzyme activity and enriching acidifying bacteria. This study demonstrated that SP-TH pretreatment can effectively promote acid production from sludge, providing a new avenue for organic matter recovery through sludge anaerobic fermentation pretreatment.

The effect of ultrasound on the extraction and foaming properties of proteins from potato trimmings.

High-intensity ultrasonication is an emerging technology for plant protein isolation and modification. In this study, the potential of temperature-controlled ultrasonication to enhance the recovery of functional proteins from potato trimmings was assessed. Different ultrasound energy levels [2000-40,000 J/g fresh weight (FW)] were applied during protein extraction at pH 9.0. True protein yields after ultrasonication significantly increased (up to 91%) compared to conventional extraction (33%). Microstructural analysis of the extraction residues showed more disrupted cells as ultrasonication time increased. Ultrasound treatments (10,000 and 20,000 J/g FW) increased the protein yield without affecting the foaming and air-water interfacial properties of protein isolates obtained after isoelectric precipitation (pH 4.0). However, proteins obtained after extended ultrasonication (40,000 J/g FW) had significantly slower early-stage adsorption kinetics. This was attributed to ultrasound-induced aggregation of the protease inhibitor fraction. In conclusion, ultrasonication shows potential to help overcome some challenges associated with plant protein extraction.

Enhancing phycocyanin yield from Spirulina sp. under salt stress using various extraction methods.

Phycocyanin, a blue-coloured pigment, predominantly found and derived from Spirulina sp., has gained researchers' interest due to its vibrant hues and other attractive properties like antioxidant and anti-microbial. However, the lack of reliable and sustainable phycocyanin extraction strategies without compromising the quality has hindered the scaling up of its production processes for commercial purposes. Here in this study, phycocyanin was extracted from wet and dry biomass Spirulina sp., using three different physical cell disruption methods (ultrasonication, homogenization, and freeze-thaw cycles) combined with two different buffers (phosphate buffer and acetate buffer) and water (as control). The result showed that the freeze-thaw method combined with acetate buffer produced the highest yield (25.013 ± 2.572 mg/100 mg) with a purity ratio of 0.806 ± 0.079. Furthermore, when subjected to 30% w/v salt stress, 1.9 times higher phycocyanin yield with a purity ratio of 1.402 ± 0.609 was achieved using the previously optimized extraction method.

Extraction of Protein and Bioactive Compounds from Mediterranean Red Algae (Sphaerococcus coronopifolius and Gelidium spinosum) Using Various Innovative Pretreatment Strategies.

In this study, the release of proteins and other biomolecules into an aqueous media from two red macroalgae (Sphaerococcus coronopifolius and Gelidium spinosum) was studied using eight different cell disruption techniques. The contents of carbohydrates, pigments, and phenolic compounds coextracted with proteins were quantified. In addition, morphological changes at the cellular level in response to the different pretreatment methods were observed by an optical microscope. Finally, the antioxidant capacity of obtained protein extracts was evaluated using three in vitro tests. For both S. coronopifolius and G. spinosum, ultrasonication for 60 min proved to be the most effective technique for protein extraction, yielding values of 3.46 ± 0.06 mg/g DW and 9.73 ± 0.41 mg/g DW, respectively. Furthermore, the highest total contents of phenolic compounds, flavonoids, and carbohydrates were also recorded with the same method. However, the highest pigment contents were found with ultrasonication for 15 min. Interestingly, relatively high antioxidant activities like radical scavenging activity (31.57-65.16%), reducing power (0.51-1.70, OD at 700 nm), and ferrous iron-chelating activity (28.76-61.37%) were exerted by the different protein extracts whatever the pretreatment method applied. This antioxidant potency could be attributed to the presence of polyphenolic compounds, pigments, and/or other bioactive substances in these extracts. Among all the used techniques, ultrasonication pretreatment for 60 min appears to be the most efficient method in terms of destroying the macroalgae cell wall and extracting the molecules of interest, especially proteins. The protein fractions derived from the two red macroalgae under these conditions were precipitated with ammonium sulfate, lyophilized, and their molecular weight distribution was determined using SDS-PAGE. Our results showed that the major protein bands were observed between 25 kDa and 60 kDa for S. coronopifolius and ranged from 20 kDa to 150 kDa for G. spinosum. These findings indicated that ultrasonication for 60 min could be sufficient to disrupt the algae cells for obtaining protein-rich extracts with promising biological properties, especially antioxidant activity.

Pulsed electric field (PEF) processing of microalga Chlorella vulgaris and its digestibility in broiler feed.

Microalgae have potentially beneficial effects on animal health and nutritional value when added to feed. Crucial hereby is that intracellular bio-active molecules are released in the intestinal tract. Digestibility of Chlorella vulgaris and its impact on total digestibility of broiler feed is a first step in assessing its characteristics as feed supplement. Different methods could be used to increase the digestibility of the algae. Among other, pulsed electric field (PEF) and freezing to disrupt autotrophic (A) and heterotrophic (H) Chlorella vulgaris cells was assessed to increase their availability followed by in-vivo trials. In these trials effect of algae type (A and H) and effect of PEF-processing was evaluated on the apparent nutrient digestibility. Pulsed electric field showed to have a disruption efficiency of 83.90% and 79.20% for heterotrophic and autotrophic C. vulgaris respectively. Freezing C. vulgaris only showed efficiencies ranging from 3.86 to 11.58%. In the in-vivo trials, microscopic counting of intact C. vulgaris cells showed an increase in digested intact C. vulgaris cells of PEF-processed C. vulgaris compared to nonprocessed cells ranging from 12.16% to 15.20%. Autotrophic C. vulgaris had a higher digestibility compared to heterotrophic C. vulgaris, with an increase of 7.29, 9.44, and 17.29% in digestibility of C. vulgaris in the 1, 2, and 5% feed respectively. Feeds with PEF-processed C. vulgaris showed no significant increase in digestibility compared to nonprocessed C. vulgaris supplemented feeds. The 5% C. vulgaris feeds showed lower fat digestibility than the 1 and 2% and control feeds. Protein digestibility was lower for all C. vulgaris feeds compared to the control feed. There was a significant linear decreasing effect (P < 0.001) for all digestibility parameters. Except for crude ash digestibility, which first lowered for the 1 and 2% feeds, but then increased at 5% inclusion. Considering this study, including low dosages of 1 and 2% of C. vulgaris in broiler feed does not compromise its digestibility.

Green and efficient method to acquire high-value phycobiliprotein from microalgal biomass involving deep eutectic solvent-based ultrasound-assisted extraction.

Phycoerythrin (PE) is a phycobiliprotein holding great potential as a high-value food colorant and medicine. Deep eutectic solvent (DES)-based ultrasound-assisted extraction (UAE) was applied to extract B-PE by disrupting the resistant polysaccharide cell wall of Porphyridium purpureum. The solubility of cell wall monomers in 31 DESs was predicted using COSMO-RS. Five glycerol-based DESs were tested for extraction, all of which showed significantly higher B-PE yields by up to 13.5 folds than water. The DES-dependent B-PE extraction efficiencies were proposedly associated with different cell disrupting capabilities and protein stabilizing effects of DESs. The DES-based UAE method could be considered green according to a metric assessment tool, AGREEprep. The crude extract containing DES was further subjected to aqueous two-phase system, two-step ammonium sulfate precipitation, and ultrafiltration processes. The final purified B-PE had a PE purity ratio of 3.60 and a PC purity ratio of 0.08, comparable to the purity of commercial products.

Novel insights into the anti-asthmatic effect of Raphanus sativus L. (Raphani Semen): Targeting immune cells, inflammatory pathways and oxidative stress markers.

ETHNOPHARMACOLOGICAL RELEVANCE: Raphanus sativus L. is a well-known medicinal plant with traditional therapeutic applications in various common ailments including inflammation and asthma. AIMS OF THE STUDY: This study aimed to evaluate the chemical composition and anti-asthmatic potential of the hydro-methanolic extract of the leaves of R. sativus L. (Rs.Cr) using various in vitro and in vivo investigations. MATERIALS AND METHODS: The Rs.Cr was subjected to preliminary phytochemical analysis and HPLC profiling. The safety was assessed through oral acute toxicity tests in mice. The antiasthmatic effect of the extract was studied using milk-induced leukocytosis and ovalbumin (OVA)-induced allergic asthma models established in mice. While mast cell degranulation and passive paw anaphylaxis models were established in rats. Moreover, effect of the extract was studied on various oxidative and inflammatory makers. The antioxidant effect of the extract was also studied by in vitro DPPH method. RESULTS: The HPLC profiling of Rs.Cr showed the presence of important polyphenols in a considerable quantity. In toxicity evaluation, Rs.Cr showed no sign of morbidity or mortality with LD50 < 2000 mg/kg. The extract revealed significant mast cell disruption in a dose-dependent manner compared to the intoxicated group. Similarly, treatment with Rs.Cr and dexamethasone significantly (p < 0.001) reduced paw edema volume. Subcutaneous injection of milk at a dose of 4 mL/kg, after 24 h of its administration, showed an increase in the leukocyte count in the intoxicated group. Similarly, mice treated with dexamethasone and Rs.Cr respectively showed a significant decrease in leukocytes and eosinophils count in the ovalbumin-induced allergic asthma model. The extract presented a significant (p˂0.001) alleviative effect on the levels of SOD and GSH, MDA, IL-4, IL-5, and IL-13 in a dose-dependent manner as compared to the intoxicated group. Furthermore, the histological evaluation also revealed a notable decrease in inflammatory and goblet cell count with reduced mucus production. CONCLUSION: The current study highlights mechanism-based novel insights into the anti-asthmatic potential of R. sativus that also strongly supports its traditional use in asthma.

Characterization of yeast protein isolates extracted via high-pressure homogenization and pH shift: A promising protein source enriched with essential amino acids and branched-chain amino acids.

In the global food industry, plant-based protein isolates are gaining prominence as an alternative to animal-based counterparts. However, their nutritional value often falters due to insufficient essential amino acids. To address this issue, our study introduces a sustainable protein isolate derived from yeast cells, achieved through high-pressure homogenization (HPH) and alkali pH-shifting treatment. Subjected to HPH pressures ranging from 60 to 120 MPa and 1 to 10 cycles, higher pressure and cycle numbers resulted in enhanced disruption of yeast cells. Combining HPH with alkali pH-shifting treatment significantly augmented protein extraction. Four cycles of HPH at 100 MPa yielded the optimized protein content, resulting in a yeast protein isolate (YPI) with 75.3 g protein per 100 g powder, including 30.0 g of essential amino acids and 18.4 g of branched-chain amino acids per 100 g protein. YPI exhibited superior water and oil-holding capacities compared to pea protein isolate, whey protein isolate (WPI), and soy protein isolate. Although YPI exhibited lower emulsifying ability than WPI, it excelled in stabilizing protein-stabilized emulsions. For foaming, YPI outperformed others in both foaming ability and stabilizing protein-based foam. In conclusion, YPI surpasses numerous plant-based protein alternatives in essential amino acids and branched-chain amino acids contents, positioning it as an excellent candidate for widespread utilization as a sustainable protein source in the food industry, owing to its exceptional nutritional advantages, as well as emulsifying and foaming properties. PRACTICAL APPLICATION: This study introduces a sustainable protein isolate derived from yeast cells. YPI exhibited considerable promise as a protein source. Nutritionally, YPI notably surpassed plant-based protein isolates in EAA and BCAA contents. Functionally, YPI demonstrated superior water-holding and oil-holding capacities, as well as an effective emulsion and foam stabilizer.

Optimization of bio-oil extraction from Chlorella biomass via a green approach to obtain algal-based Di-ethyl phthalate.

This study focuses on determining the optimum external operating parameters of algal cell lysis for extraction of bio-oil from Chlorella biomass. Response surface methodology has been applied to a regression analysis model for optimizing solvent ratios, i.e., ethyl acetate to ethanol (E.A.:E) ratio for maximum extraction of bio-oil and aqueous deep eutectic solvent to biomass (aDES:biomass) ratio for algal pretreatment for the enhanced yield of bio-oil. Optimized process conditions were 15 min of homogenization combined with ultrasonication (hybrid method). The aDES:biomass ratio of 8.25 caused the highest cell disruption efficiency to liberate bio-oil from encapsulated cells. The solvent ethyl acetate to ethanol ratio (E.A.:E) was optimum at 0.8 for maximum extraction of bio-oil, and studies indicated a maximum bio-oil yield of 94.0% using this hybrid pretreatment process combined with ultrasonication and homogenization. The GC-MS characterization technique was used to analyze the bio-oil, which showed it consisted of 67.93% Di-ethyl phthalate (DEP) and 32.07% esters compounds (C12-C40 hydrocarbons range). The produced DEP from Chlorella biomass using this sustainable green approach is very promising. The estimated cost was around Rs 49 per gm (equivalent to Rs 664.56 for 13.58 gm), which indicates the potential for a cost-effective method to produce pure DEP from Chlorella biomass.

Phycobiliproteins from microalgae: research progress in sustainable production and extraction processes.

Phycobiliproteins (PBPs), one of the functional proteins from algae, are natural pigment-protein complex containing various amino acids and phycobilins. It has various activities, such as anti-inflammatory and antioxidant properties. And are potential for applications in food, cosmetics, and biomedicine. Improving their metabolic yield is of great interest. Microalgaes are one of the important sources of PBPs, with high growth rate and have the potential for large-scale production. The key to large-scale PBPs production depends on accumulation and recovery of massive productive alga in the upstream stage and the efficiency of microalgae cells breakup and extract PBPs in the downstream stage. Therefore, we reviewed the status quo in the research and development of PBPs production, summarized the advances in each stage and the feasibility of scaled-up production, and demonstrated challenges and future directions in this field.

Development of sustainable downstream processing for nutritional oil production.

Nutritional oils (mainly omega-3 fatty acids) are receiving increased attention as critical supplementary compounds for the improvement and maintenance of human health and wellbeing. However, the predominant sources of these oils have historically shown numerous limitations relating to desirability and sustainability; hence the crucial focus is now on developing smarter, greener, and more environmentally favourable alternatives. This study was undertaken to consider and assess the numerous prevailing and emerging techniques implicated across the stages of fatty acid downstream processing. A structured and critical comparison of the major classes of disruption methodology (physical, chemical, thermal, and biological) is presented, with discussion and consideration of the viability of new extraction techniques. Owing to a greater desire for sustainable industrial practices, and a desperate need to make nutritional oils more available; great emphasis has been placed on the discovery and adoption of highly sought-after 'green' alternatives, which demonstrate improved efficiency and reduced toxicity compared to conventional practices. Based on these findings, this review also advocates new forays into application of novel nanomaterials in fatty acid separation to improve the sustainability of nutritional oil downstream processing. In summary, this review provides a detailed overview of the current and developing landscape of nutritional oil; and concludes that adoption and refinement of these sustainable alternatives could promptly allow for development of a more complete 'green' process for nutritional oil extraction; allowing us to better meet worldwide needs without costing the environment.

Antibacterial activity of peptides and bio-safety evaluation: in vitro and in vivo studies against bacterial and fungal pathogens.

BACKGROUND: Antimicrobial peptides are promising alternatives to antibiotics to treat bacterial and fungal infections, especially drug-resistant clinical pathogens. METHODS: Antimicrobial peptides (AMPs) were synthesized and antimicrobial activity was assayed. The antibacterial mechanism, ATP production, ROS generation and molecular mechanism were determined. Biofilm inhibition assay was performed in planktonic bacterial cells and biofilm degradation assay was performed using mature biofilm. The synthesized AMP2 was subjected to in vitro and in vivo analysis to analyze the safety. RESULTS: The synthesized peptides AMP1, AMP2, AMP3 and AMP4 exhibited antimicrobial activity against Gram-positive and Gram-negative bacteria. The MIC values ranged from 1.5 ± 0.25-12.5 ± 1.25 µM and the MFC values range from 2.25 ± 0.12-25 ± 1.25 µM. F. solani showed fewer MFC values than other fungal strains. Time kill assay was performed and the AMP2 killed about 70 % of Acinetobacter baumannii at 1 × MIC concentration within 10 min incubation and killed 97 % of bacteria at 1 × MBC concentration within 15 min. The antimicrobial peptide AMP2 was highly effective against planktonic A. baumannii and L. monocytogenes. The tested AMP2 showed less toxicity to cell lines and Zebrafish. CONCLUSIONS: Antimicrobial peptides have potential antimicrobial properties against Gram-positive and Gram-negative bacteria. The in silico studies of these antimicrobial peptides are useful for eradicating drug-resistant bacteria.

Comparative Analysis of Volatile Compounds from Four Radish Microgreen Cultivars Based on Ultrasonic Cell Disruption and HS-SPME/GC-MS.

The ultrasonic cell disruption method was used to efficiently extract isothiocyanates and other volatile compounds from radish microgreens. A total of 51 volatiles were identified and quantified by headspace solid-phase micro-extraction and gas chromatography-mass spectrometry (HS-SPME/GC-MS) in four radish microgreen cultivars, mainly including alcohols, aldehydes, isothiocyanates, sulfides, ketones, esters, terpenes, and hydrocarbons. The correlation between cultivars and volatile compounds was determined by chemometrics analysis, including principal component analysis (PCA) and hierarchical clustering heat maps. The aroma profiles were distinguished based on the odor activity value (OAV), odor contribution rate (OCR), and radar fingerprint chart (RFC) of volatile compounds. This study not only revealed the different flavor characteristics in four cultivars but also established a theoretical basis for the genetic improvement of radish microgreen flavors.

Numerical investigation of force and deflection of nanoneedle penetration into cell using finite element approach: Parameter study and experimental validation of results.

This paper aims to develop a numerical methodology to investigate the penetration process of nanoneedles into cells and the corresponding force and indentation length. The finite element approach via the explicit dynamic method handles convergence difficulties in the nonlinear phenomenon. The cell is modeled as an isotropic elastic hemiellipsoidal shell with a thickness of 200 nm, which represents the lipid membrane and actin cortex, encapsulating cytoplasm that is regarded as an Eulerian body because of its fluid-type behavior. Nanoneedles with diameters 400, 200, and 50 nm are considered for model development based on available experimental data. The Von Mises strain failure criterion is used for rupture detection. A parameter study using 1, 2.5, 5, 7.5, and 10 kPa shows that Young's modulus of the HeLa cell membrane is about 5 kPa. Moreover, a failure strain of 1.2 chosen among 0.2, 0.4, 0.6, 0.8, 1, and 1.2 matches best the experimental data. In addition, a diameter study shows that the relations between force-diameter and indentation length-diameter are linear and polynomial, respectively. Furthermore, regarding the experimental data and by using contour of minimum principal stress around needle and an analytical equation for calculation of buckling force of a woven fabric, we proposed that for a given cell, membrane structural stability-a function of the coupled effect of Young's modulus and actin meshwork size-contributes directly to needle insertion success rate for that type of cell.

Non-thermal, energy efficient hydrodynamic cavitation for food processing, process intensification and extraction of natural bioactives: A review.

Hydrodynamic cavitation (HC) is the process of bubbles formation, expansion, and violent collapse, which results in the generation of high pressures in the order of 100-5000 bar and temperatures in the range of 727-9727 °C for just a fraction of seconds. Increasing consumer demand for high-quality foods with higher nutritive values and fresh-like sensory attributes, food processors, scientists, and process engineers are pushed to develop innovative and effective non-thermal methods as an alternative to conventional heat treatments. Hydrodynamic cavitation can play a significant role in non-thermal food processing as it has the potential to destroy microbes and reduce enzyme activity while retaining essential nutritional and physicochemical properties. As hydrodynamic cavitation occurs in a flowing liquid, there is a decrease in local pressure followed by its recovery; hence it can be used for liquid foods. It can also be used to create stable emulsions and homogenize food constituents. Moreover, this technology can extract food constituents such as polyphenols, essential oils, pigments, etc., via biomass pretreatment, cell disruption for selective enzyme release, waste valorization, and beer brewing. Other applications related to food production include water treatment, biodiesel, and biogas production. The present review discusses the application of HC in the preservation, processing, and quality improvement of food and other related applications. The reviewed examples in this paper demonstrate the potential of hydrodynamic cavitation with further expansion toward the scaling up, which looks at commercialization as a driving force.

Salting-out extraction of recombinant κ-carrageenase and phage T7 released from Escherichia coli cells.

Traditional technology of cell disruption has become one of the bottlenecks restricting the industrialization of genetic engineering products due to its high cost and low efficiency. In this study, a novel bioprocess of phage lysis coupled with salting-out extraction (SOE) was evaluated. The lysis effect of T7 phage on genetically engineered Escherichia coli expressing κ-carrageenase was investigated at different multiplicity of infection (MOI), meanwhile the phage and enzyme released into the lysate were separated by SOE. It was found that T7 phage could lyse 99.9% of host cells at MOI = 1 and release more than 90.0% of enzyme within 90 min. After phage lysis, 87.1% of T7 phage and 71.2% of κ-carrageenase could be distributed at the middle phase and the bottom phase, respectively, in the SOE system composed of 16% ammonium sulfate and 20% ethyl acetate (w/w). Furthermore, κ-carrageenase in the bottom phase could be salted out by ammonium sulfate with a yield of 40.1%. Phage lysis exhibits some advantages, such as mild operation conditions and low cost. While SOE can efficiently separate phage and intracellular products. Therefore, phage lysis coupled with SOE is expected to become a viable alternative to the classical cell disruption and intracellular product recovery.

Solid-state fermentation-based enzyme-assisted extraction of eicosapentaenoic acid-rich oil from Nannochloropsis sp.

Enzymatic treatment of microalgal biomass is a promising approach for extraction of microalgal lipid, but high cost of commercially sourcing enzyme is a major drawback in industrial implementation. Present study involves extraction of eicosapentaenoic acid-rich oil from Nannochloropsis sp. biomass using low cost cellulolytic enzymes produced from Trichoderma reesei in a solid-state fermentation bioreactor. Maximum total fatty acid recovery of 369.4 ± 4.6 mg/g dry weight (total fatty acid yield of 77%) was achieved in 12 h from the enzymatically treated microalgal cells, of which the eicosapentaenoic acid content was 11%. Sugar release of 1.70 ± 0.05 g/L was obtained post enzymatic treatment at 50 °C. The enzyme was reused thrice for cell wall disruption without compromising on total fatty acid yield. Additionally, high protein content of 47% in the defatted biomass could be explored as a potential aquafeed, thus enhancing the overall economics and sustainability of the process.

Alkyl carbamate ionic liquids for permeabilization of microalgae biomass to enhance lipid recovery for biodiesel production.

Microalgae are potential biomass source for biodiesel production. However, their strong cell walls make efficient lipid extraction problematic. Disrupting the cell wall is a key point in enhancing lipid yield from microalgae biomass. A new type of ionic liquid (IL) has been suggested in this work as a potentially viable solvent to permeabilize the strong microalgae cell structure for the efficient extraction of lipids. Morphological changes in microalgae cells were studied before and after ionic liquid permeabilization to understand the mechanism of ionic liquid treatment. Among the three selected CO2-based alkyl carbamate ionic liquids, DIMCARP performed with the best extraction efficiency. The effects of extraction variables (temperature, time, ratio ionic liquid/Methanol, and solvent to biomass) on lipid extraction were examined via single-factor experiments coupled with response surface methodology (RSM) using a Box-Behnken design (BBD). The highest lipid yield (16.40%) was obtained after 45 min of extraction at 45 °C using a 9:1 ionic liquid to methanol and 7 mL of solvent to biomass ratio. Transesterification of lipids to make fatty acid methyl esters found that the most common fatty acids were C16:0, C18:2, and C18:3 (19.50%). The quality of the biodiesel made meets European and US standards.

Bacterial Carotenoids: Extraction, Characterization, and Applications.

Natural carotenoids are secondary metabolites that exhibit antioxidant, anti-inflammatory, and anti-cancer properties. These types of compounds are highly demanded by pharmaceutical, cosmetic, nutraceutical, and food industries, leading to the search for new natural sources of carotenoids. In recent years, the production of carotenoids from bacteria has become of great interest for industrial applications. In addition to carotenoids with C40-skeletons, some bacteria have the ability to synthesize characteristic carotenoids with C30-skeletons. In this regard, a great variety of methodologies for the extraction and identification of bacterial carotenoids has been reported and this is the first review that condenses most of this information. To understand the diversity of carotenoids from bacteria, we present their biosynthetic origin in order to focus on the methodologies employed in their extraction and characterization. Special emphasis has been made on high-performance liquid chromatography-mass spectrometry (HPLC-MS) for the analysis and identification of bacterial carotenoids. We end up this review showing their potential commercial use. This review is proposed as a guide for the identification of these metabolites, which are frequently reported in new bacteria strains.