Green Downstream Processing Method for Xylooligosaccharide Purification and Assessment of Its Prebiotic Properties.

Xylooligosaccharides (XOS) obtained from lignocellulosic biomass after autohydrolysis primarily consist of lignin-derived impurities and autogenerated inhibitors like furfural, hydroxymethylfurfural, and acetic acid. In this study, graphene oxide-mediated purification (GOMP), a novel and environmentally friendly downstream processing method, was developed for the purification of XOS from hydrolysate obtained after ozone-assisted autohydrolysis of wheat bran. GOMP resulted in appreciable recovery of total XOS from the hydrolysate (73.87 ± 4.25%, DP2-6) with near complete removal of autogenerated inhibitors (furfural 85.42%, HMF 87.38%, and acetic acid 84.0%). Recovery of XOS by GOMP was higher than the conventional membrane purification technique (44.07 ± 0.92%) and activated charcoal treatment (72.76 ± 0.84%) along with comparatively higher removal of inhibitor compounds. GOMP results in the selective adsorption of inhibitors on the graphene oxide matrix from the XOS-rich hydrolysate, resulting in its purification and concentration. The prebiotic function of the obtained XOS fractions (DP2-4.48%, DP3-39.69%, DP4-36.13%, DP5-8.38%, and DP6-13.10%) was evaluated, indicating the growth stimulation of tested probiotic cultures and differential utilization of XOS oligomers DP3 and DP4 and complete consumption of DP2, DP5, and DP6 along with short-chain fatty acids as a major fermentation product. These findings suggest that GOMP, which employs a common substance (i.e., graphene oxide) used in water treatment, exhibits potential as an efficient and economically viable single-step methodology for XOS purification.

Food-grade xylitol production from corncob biomass with acute oral toxicity studies.

Xylitol, a sugar substitute, is widely used in various food formulations and finds a steady global market. In this study, xylitol crystals were produced from corncob by fermentation (as an alternative to the chemical catalytic process) by a GRAS yeast Pichia caribbica MTCC 5703 and characterized in detail for their purity and presence of any possible contaminant that may adversely affect mammalian cell growth and proliferation. The acute and chronic oral toxicity trials demonstrated no gross pathological changes with average weekly weight gain in female Wistar rats at high xylitol loading (LD50 > 10,000 mg/kg body weight). The clinical chemistry analysis supported the evidence of no dose-dependent effect by analyzing blood biochemical parameters. The finding suggests the possible application of the crystals (> 98% purity) as a food-grade ingredient for commercial manufacture pending human trials.

Water footprint and wastewater quality assessment of yeast single cell oil production: Gate to gate approach for industrial water sustainability.

Effective water resource utilization and sustainability for industrial operations is a growing concern. With increased industrial water demand, abstraction and water quality changes are rising. In India, distilleries generate more than 40.4 billion litres of effluent daily within the fermentation industry. Water, a public good with market and opportunity costs, needs effective mapping and management. Emerging distillery processes such as yeast lipid fermentation, if developed along with water sustainability, could aid in advancing water resource management. In the scope of this idea, the present study focuses on assessing the water footprint and water quality mapping for Rhodotorula mucilaginosa IIPL32 lipid production using crude glycerol, a by-product of the biodiesel industry. The assessment was based on primary data generated during the 500 L plant scale operation. The process's blue water footprint was assessed by applying a chain-summation approach, and the grey water requirement was determined by measuring water quality parameters for the effluent streams. The process's net blue and grey water footprint were estimated to be 3.87 and 23.66 m3 water/kg of lipid, respectively. Water quality index ratings were identified for all the respective water streams within the processing system, and human risk factors were estimated. The results suggested proper treatment of the spent broth, whereas the secondary effluent stream from cleaning operations could be reutilized within the system. Quality mapping also suggested that the effluent's high organic and mineral load can be processed for water and material recovery, which may significantly reduce the process's grey water and pollution load.

Volatile organic compounds involved in the communication of microalgae-bacterial association extracted through Headspace-Solid phase microextraction and confirmed using gas chromatography-mass spectrophotometry.

In the present study, bacterial mixture (Rhizobium and Agrobacterium) and axenic Chlorella were cultivated individually, in a mixed (co-cultured) form, and through headspace connections to study volatile organic compounds (VOCs) profile and their effect on growth. Results indicated that VOCs produced by the axenic microalgae and microalgae co-cultured with bacteria were significantly different. Axenic Chlorella predominantly produced a flavouring organic compound 2-pentadecanone (69.54%), bacterial mixed culture produced 1-decanone, 1,2,3-butanetriol, and quinoline (15-20%), and direct co-culturing of Chlorella with bacteria predominantly produced 2-pentadecanone (32.4 %). When they were allowed to communicate distantly through headspace connection, highly diversified VOCs in large numbers but low quantities were noted, predominantly 1,2-propanediol (28.82 %). In addition, growth of the co-cultured Chlorella was 1.5 times higher, while Chlorella in headspace connection with bacterial mixture exhibited âˆ¼ 3.2 times increase in growth compared to the axenic Chlorella, indicating the essential role of VOCs in growth and communication.

Unraveling of Chlorella-associated bacterial load, diversity, and their imputed functions at high- and low-yield conditions through metagenome sequencing.

Chlorella-associated bacteria can have a significant influence on facilitating higher Chlorella biomass yield due to their symbiotic relationship. In this study, non-axenic Chlorella was cultivated in an airlift photobioreactor at high and low-yield conditions. The associated bacterial diversity was analyzed using 16S rRNA metagenome sequencing. At high-yield conditions, the bacterial load was observed in the range of 108 -1010 CFU · mL-1 , whereas at low-yield conditions, bacteria were more dominant and observed in the range of 1014 -1015 CFU · mL-1 . The majority of the bacterial species associated with Chlorella at high-yield conditions belongs to Proteobacteria and Bacteroidetes. Further, Bacteroidetes levels were decreased at low-yield conditions and were highly diversified with Planctomycetes, Firmicutes, and 18 others. Predicted functional genes indicated that Chlorella-associated bacteria have the enzymes involved in the metabolism and biosynthesis of B-complex vitamins (i.e., vitamin B12 , thiamin, biotin, pyridoxine, and riboflavin). A critical evaluation revealed that vitamin biosynthesis genes were more abundant at low-yield conditions; however, vitamin B12 transport genes (B12 transport ATP-binding protein, B12 substrate-binding transportation, and B12 permease protein) were less abundant, indicating even though vitamins production occurs, but their availability to Chlorella was limited due to the lack of vitamin transport genes. Further, at high yield, Chlorella-associated bacteria enabled higher growth by supplementing the vitamins. In contrast, at low-yield condition-an increased bacterial load, diversity, and limited vitamin transport functional genes affected the Chlorella yield. It can be inferred that Chlorella yield was significantly affected by three factors: associated bacterial load, diversity, and transport functional genes of vitamins.

Salt Stress-Induced Anthocyanin Biosynthesis Genes and MATE Transporter Involved in Anthocyanin Accumulation in Daucus carota Cell Culture.

Anthocyanins biosynthesis is a well-studied biosynthesis pathway in Daucus carota. However, the scale-up production at the bioreactor level and transporter involved in accumulation is poorly understood. To increase anthocyanin content and elucidate the molecular mechanism involved in accumulation, we examined D. carota cell culture in flask and bioreactor for 18 days under salt stress (20.0 mM NH4NO3/37.6 mM KNO3) at 3 day intervals. The expression of anthocyanin biosynthesis and putative MATE (multidrug and toxic compound extrusion) transporter expression was analyzed by qRT-PCR. It was observed that there was a significant enhancement of anthocyanin in the bioreactor compared to the control culture. A correlation was observed between the expression of MATE and the anthocyanin biosynthesis genes (CHS, C4H, LDOX, and UFGT) on the 9th day in a bioreactor, where maximum anthocyanin accumulation and expression was detected. We hypothesize the involvement of MATE in transporting anthocyanin to tonoplast in D. carota culture under salt stress.

Ozone assisted autohydrolysis of wheat bran enhances xylooligosaccharide production with low generation of inhibitor compounds: A comparative study.

In the present study, ozone assisted autohydrolysis (OAAH) was evaluated for enhanced generation of xylooligosaccharide (XOS) from wheat bran. The total XOS yield with optimum ozone dose of 3% (OAAH-3) was found to be 8.9% (w/w biomass) at 110 °C in comparison to 7.96% at 170 °C by autohydrolysis (AH) alone. Although, there was no significant difference in oligomeric composition (DP 2-6), significant decrease in degradation products namely furfural (2.78-fold), HMF (3.15-fold), acrylamide (nil) and acetic acid (1.06-fold), was observed with OAAH-3 as a pretreatment option. There was 1-fold higher xylan to XOS conversion and OAAH-hydrolysate had higher DPPH radical scavenging activity than AH. PCA plots indicated clear enhancement in XOS production and lower generation of inhibitors with decrease in treatment temperature. Results of the study therefore suggest OAAH can be an effective pretreatment option that can further be integrated with downstream processing for concentration and purification of XOS.

Targeting COVID-19 (SARS-CoV-2) main protease through active phytocompounds of ayurvedic medicinal plants - Emblica officinalis (Amla), Phyllanthus niruri Linn. (Bhumi Amla) and Tinospora cordifolia (Giloy) - A molecular docking and simulation study.

Coronavirus Disease-2019 (COVID-19), a viral disease caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) was declared a global pandemic by WHO in 2020. In this scenario, SARS-CoV-2 main protease (COVID-19 Mpro), an enzyme mainly involved in viral replication and transcription is identified as a crucial target for drug discovery. Traditionally used medicinal plants contain a large amount of bioactives and pave a new path to develop drugs and medications for COVID-19. The present study was aimed to examine the potential of Emblica officinalis (amla), Phyllanthus niruri Linn. (bhumi amla) and Tinospora cordifolia (giloy) bioactive compounds to inhibit the enzymatic activity of COVID-19 Mpro. In total, 96 bioactive compounds were selected and docked with COVID-19 Mpro and further validated by molecular dynamics study. From the docking and molecular dynamics study, it was revealed that the bioactives namely amritoside, apigenin-6-C-glucosyl7-O-glucoside, pectolinarin and astragalin showed better binding affinities with COVID-19 Mpro. Drug-likeness, ADEMT and bioactivity score prediction of best drug candidates were evaluated by DruLiTo, pkCSM and Molinspiration servers, respectively. Overall, the in silico results confirmed that the validated bioactives could be exploited as promising COVID-19 Mpro inhibitors.

Chlorella vulgaris cultivation in airlift photobioreactor with transparent draft tube: effect of hydrodynamics, light and carbon dioxide on biochemical profile particularly ω-6/ω-3 fatty acid ratio.

Chlorella vulgaris is used for food and feed applications due to its nutraceutical, antioxidant and anticancer properties. An airlift photobioreactor comprising transparent draft tube was used for C. vulgaris cultivation. The effect of reactor parameters like hydrodynamics (0.3-1.5 vvm), light intensity (85-400 µmol m-2 s-1), photoperiod (12-24 h) and gas-phase carbon dioxide (CO2) concentration (5-15% v/v) were evaluated on microalgae and associated bacterial growth, biochemical profile; with special emphasis on ω-3, ω-6 fatty acids, and vitamin B12. The optimal growth of C. vulgaris without CO2 supplementation was observed at 1.2 vvm, which was associated with higher algal productivity, chlorophyll, vitamin B12 content, and bacterial load along with 72% of nitrate removal. The higher light intensity (400 µmol m-2 s-1) and photoperiod (24:0) increased biomass productivity and ω-3 fatty acid content (in lipid) up to 2-3 fold. The elevated levels of gas-phase CO2 concentration (15% v/v) enhanced EPA content up to 7% and biomass productivity up to 171 mg L-1 day-1. However, the increase in CO2 concentration lowered vitamin B12 content (up to 30%) and bacterial load (2-3 log). Also, all the cultivation conditions favoured desirable ω-6/ω-3 ratio(in the range of 1-2).

Comparative life cycle assessment of autotrophic cultivation of Scenedesmus dimorphus in raceway pond coupled to biodiesel and biogas production.

Life cycle assessment (LCA) of indigenous freshwater microalgae, Scenedesmus dimorphus, cultivation in open raceway pond and its conversion to biodiesel and biogas were carried out. The LCA inventory inputs for the biogas scenario was entirely based on primary data obtained from algal cultivation (in pilot scale raceway pond), harvesting, and biogas production; while only the downstream processing involved in biodiesel production namely drying, reaction and purification were based on secondary data. Overall, eight scenarios were modeled for the integrated process involving: algae-based CO2 capture and downstream processing scenarios for biodiesel and biogas along with impact assessment of nutrient addition and extent of recycling in a life cycle perspective. The LCA results indicated a huge energy deficit and net CO2 negative in terms of CO2 capture for both the biodiesel and biogas scenarios, majorly due to lower algal biomass productivity and higher energy requirements for culture mixing. The sensitivity analysis indicated that variability in the biomass productivity has predominant effect on the primary energy demand and global warming potential (GWP, kg CO2 eq.) followed by specific energy consumption for mixing algal culture. Furthermore, the LCA results indicated that biogas conversion route from microalgae was more energy efficient and sustainable than the biodiesel route. The overall findings of the study suggested that microalgae-mediated CO2 capture and conversion to biodiesel and biogas production can be energy efficient at higher biomass productivity (> 10 g m-2 day-1) and via employing energy-efficient systems for culture mixing (< 2 W m-3).

Development of a carotenoid enriched probiotic yogurt from fresh biomass of Spirulina and its characterization.

Incorporation of Spirulina in milk as thermally dried powder has the disadvantages of non-uniform distribution with undesirable odor and flavor. Through homogenization (200 ± 10 bar), complete dispersion of fresh Spirulina biomass (7% w/w) in milk was achieved and thereafter a carotenoid enriched probiotic yogurt was developed. Confocal microscopy revealed porous Spirulina-milk protein matrix integrated with smaller fat globules in the yogurt. Spirulina led to a 29.56% increase in Lactobacillus acidophilus count, a 20% reduction in fermentation time and a total probiotic count of 1.2 × 107 CFU mL-1. The protein, total chlorophyll, total carotenoid and ß-carotene content (on dry w/w basis) were 3.58 ± 0.08 g 100 g-1, 0.407 ± 0.018 mg g-1, 0.235 ± 0.016 mg g-1 and 13.28 ± 0.08 µg g-1, respectively. During storage (18 days at 6-8 °C), the L. acidophilus count reached 8.83 ± 0.11 log CFU mL-1 with 103.03% increase in the viability by day three and the yogurt retained 71.5% carotenoids. The probiotc Spirulina yogurt was found to be acceptable to consumers as evaluated by affective consumer test.

Ozone pre-treatment of molasses-based biomethanated distillery wastewater for enhanced bio-composting.

Composting is a biological process in which the organic matter is degraded by the mixed population of microorganisms in a moist aerobic environment to more stable and humidified end products. The composting process involves an interaction between the organic waste, microorganisms, moisture and oxygen. The molasses-based biomethanated distillery wastewater is presently effectively utilized with sugarcane pressmud through the composting process. The aim of present work was to evaluate the effect of ozone pretreatment on the rate of composting process and the quality of compost obtained. The GC-MS & FTIR analysis of ozone pretreated wastewater indicated the degradation and/or transformation of the organic compounds to simpler compounds present in the wastewater. Composting was performed by mixing fixed weight ratios of pressmud and different ratios of ozone pretreated wastewater (1:3, 1:4 and 1:5). The composting process was found to occur faster in the ozone pretreated wastewater for all the ratios as compared to the untreated wastewater. The final compost characteristics were found to be optimum for the 1:3 and 1:4 ratios of pressmud and wastewater. The bio-oxidative phase duration of composting has been reduced for the ozone pretreated wastewater.

Treatment of pharmaceutical industrial wastewater by nano-catalyzed ozonation in a semi-batch reactor for improved biodegradability.

The study reports the biodegradability enhancement of pharmaceutical wastewater along with COD (Chemical Oxygen Demand) color and toxicity removal via O3, O3/Fe2+, O3/nZVI (nano zero valent iron) processes. Nano catalytic ozonation process (O3/nZVI) showed the highest biodegradability (BI = BOD5/COD) enhancement of pharmaceutical wastewater up to 0.63 from 0.18 of control with a COD, color and toxicity removal of 62.3%, 93% and 82% respectively. The disappearance of the corresponding Fourier transform infrared (FTIR) and gas chromatography-mass spectrometry (GC-MS) peaks after pretreatment indicated the degradation or transformation of the refractory organic compounds to more biodegradable organic compounds. The subsequent aerobic degradation study of pretreated pharmaceutical wastewater resulted in biodegradation rate enhancement of 5.31, 2.97, and 1.22 times for O3/nZVI O3/Fe2+ and O3 processes respectively. Seed germination test using spinach (Spinacia oleracea) seeds established the toxicity removal of pretreated pharmaceutical wastewater.

Nano catalytic ozonation of biomethanated distillery wastewater for biodegradability enhancement, color and toxicity reduction with biofuel production.

The effectiveness of O3, O3/Fe2+, and O3/nZVI processes on biomethanated distillery wastewater (BMDWW) was evaluated in terms of biodegradability index (BI) enhancement, biofuel production, COD, color & toxicity reduction. A significant increase in biodegradability, COD, color and toxicity reduction was observed in O3/nZVI compared with O3, O3/Fe2+ due to more hydroxyl radical production. The O3/nZVI pretreated wastewater with enhanced BI (up to 0.71) showed 60% COD removal with additional biogas generation (64% methane content). From the Gas Chromatography Mass Spectrometry (GC-MS) analysis, 18 foremost organic compounds were predominantly detected in the raw distillery wastewater. The disappearance of the corresponding FTIR (Fourier Transform Infrared Spectroscopy) & GC-MS spectra during pretreatment processes signified the degradation or transformation of the recalcitrant present in the distillery wastewater. Subsequent (AnO + AO, AO) of pretreated BMDWW resulted in biodegradation rate enhancement by (1.83, 1.67), (3.5, 2.4) and (4.7, 2.9) times for O3, O3/Fe2+ and O3/nZVI processes respectively.

Recent Advances in Microalgal Bioactives for Food, Feed, and Healthcare Products: Commercial Potential, Market Space, and Sustainability.

To combat food scarcity as well as to ensure nutritional food supply for sustainable living of increasing population, microalgae are considered as innovative sources for adequate nutrition. Currently, the dried biomass, various carotenoids, phycocyanin, phycoerythrin, omega fatty acids, and enzymes are being used as food additives, food coloring agents, and food supplements. Apart from nutritional importance, microalgae are finding the place in the market as "functional foods." When compared to the total market size of food and feed products derived from all the possible sources, the market portfolio of microalgae-based products is still smaller, but increasing steadily. On the other hand, the genetic modification of microalgae for enhanced production of commercially important metabolites holds a great potential. However, the success of commercial application of genetically modified (GM) algae will be defined by their safety to human health and environment. In view of this, the present study attempts to highlight the industrially important microalgal metabolites, their production, and application in food, feed, nutraceuticals, pharmaceuticals, and cosmeceuticals. The current and future market trends for microalgal products have been thoroughly discussed. Importantly, the safety pertaining to microalgae cultivation and consumption, and regulatory issues for GM microalgae have also been covered.

Catalytic ozone pretreatment of complex textile effluent using Fe2+ and zero valent iron nanoparticles.

The study investigates the effect of catalytic ozone pretreatment via Fe2+ and nZVI on biodegradability enhancement of complex textile effluent. The nZVI particles were synthesized and characterized by XRD, TEM and SEM analyses. Results showed that nano catalytic ozone pretreatment led to higher biodegradability index (BOD5/COD = BI) enhancement up to 0.61 (134.6%) along with COD, color and toxicity removal up to 73.5%, 87%, and 92% respectively. The disappearance of the corresponding GCMS & FTIR spectral peaks during catalyzed ozonation process indicated the cleavage of chromophore group and degradation of organic compounds present in the textile effluent. Subsequent aerobic biodegradation of nZVI pretreated textile effluent resulted in maximum COD and color reduction of 78% and 98.5% respectively, whereas the untreated effluent (BI = 0.26) indicated poor COD and color reduction of only 31% and 33% respectively. Bio-kinetic parameters also confirmed the increased rate of bio-oxidation at enhanced BIs. Seed germination test using seeds of Spinach (Spinacia oleracea), indicated the effectiveness of nano catalyzed ozone pretreatment in removing toxicity from contaminated textile effluent.

Comparison of coagulation, ozone and ferrate treatment processes for color, COD and toxicity removal from complex textile wastewater.

In this study, the comparative performance of coagulation, ozone, coagulation + ozone + coagulation and potassium ferrate processes to remove chemical oxygen demand (COD), color, and toxicity from a highly polluted textile wastewater were evaluated. Experimental results showed that ferrate alone had no effect on COD, color and toxicity removal. Whereas, in combination with FeSO4, it has shown the highest removal efficiency of 96.5%, 83% and 75% for respective parameters at the optimal dose of 40 mgL-1 + 3 ml FeSO4 (1 M) in comparison with other processes. A seed germination test using seeds of Spinach (Spinacia oleracea) also indicated that ferrate was more effective in removing toxicity from contaminated textile wastewater. Potassium ferrate also produces less sludge with maximum contaminant removal, thereby making the process more economically feasible. Fourier transform infrared spectroscopy (FTIR) analysis also shows the cleavage of the chromophore group and degradation of textile wastewater during chemical and oxidation treatment processes.

Biomass and lipid enhancement in Chlorella sp. with emphasis on biodiesel quality assessment through detailed FAME signature.

In this study, the concentrations of MgSO4, salinity and light intensity were optimised for maximum biomass productivity and lipid content in Chlorella sp. Lipid synthesized at varied experimental conditions was also assessed in detail for biodiesel properties through FAME analysis. FAMEs mainly composed of C16:0, C16:1(9), C16:3(7, 10, 13), C18:0, C18:1(11), C18:2(9, 12), C18:3(9, 12, 15). The optimum biomass productivity (372.50mgL(-1)d(-1)) and lipid content (32.57%) was obtained at MgSO4-150ppm; salinity-12.5ppm, and light intensity-25µmolm(-2)s(-1). However, at this condition the cetane number, a major biodiesel property was not complying with worldwide biodiesel standard. Therefore, further optimisations were done to check the suitability of biodiesel fuel. The optimum biomass productivity (348.47mgL(-1)d(-1)) and lipid content (12.43%) with suitable biodiesel fuel properties was obtained at MgSO4-50ppm, salinity-25ppm and light intensity-100µmolm(-2)s(-1). The validation experiments confirmed the closeness of predicted and measured response values.

Examination of Lead and Cadmium in Water-based Paints Marketed in Nigeria.

BACKGROUND: In spite of the availability of substitutes for lead and cadmium compounds in paints, manufacturers continue to produce paints with high levels of these metals. As the population continues to grow and there is a continued shift from oil-based to water-based paints, the sales and use of these paints will increase the exposure of humans and the environment to these metals. OBJECTIVES: We measured the levels of lead (Pb) and cadmium (Cd) in 174 paint samples marketed in Lagos and Ibadan, Nigeria. Paint samples from different manufacturers registered with and without Standards Organization of Nigeria (SON) were considered. METHODS: Samples were acid digested using a microwave digester and the levels of the elements were determined using inductively coupled plasma optical emission spectroscopy (ICP-OES). DISCUSSION: The levels of Cd and Pb (dry weight) in all samples ranged from 98-1999 µg/g and 170-3231 µg/g, respectively. All the samples were above the permissible limits of 90 ppm of the US Consumer Product Safety Commission and 100 ppm limit of the European Union (EU) for Pb and Cd in paint. CONCLUSIONS: We concluded that water-based paints marketed in Nigeria still contain substantial amounts of lead and cadmium which are detrimental to human health and the entire ecosystem. These metals are among the EU priority metals due to the increased risk of occupational exposure to humans and vulnerable groups such as children.

Carbonic anhydrase mediated carbon dioxide sequestration: promises, challenges and future prospects.

Anthropogenic activities have substantially increased the level of greenhouse gases (GHGs) in the atmosphere and are contributing significantly to the global warming. Carbon dioxide (CO2 ) is one of the major GHGs which plays a key role in the climate change. Various approaches and methodologies are under investigation to address CO2 capture and sequestration worldwide. Carbonic anhydrase (CA) mediated CO2 sequestration is one of the promising options. Therefore, the present review elaborates recent developments in CA, its immobilization and bioreactor methodologies towards CO2 sequestration using the CA enzyme. The promises and challenges associated with the efficient utilization of CA for CO2 sequestration and scale up from flask to lab-scale bioreactor are critically discussed. Finally, the current review also recommends the possible future needs and directions to utilize CA for CO2 sequestration.