Advances in microalgal research for valorization of industrial wastewater.

This review article focuses on recent updates on remediation of industrial wastewater (IWW) through microalgae cultivation. These include how adding additional supplements of nutrient to some specific IWWs lacking adequate nutrients improving the microalgae growth and remediation simultaneously. Various pretreatments strategy recently employed for IWWs treatment other than dealing with microalgae was discussed. Various nutrient-rich IWW could be utilized directly with additional dilution, supplement of nutrients and without any pretreatment. Recent advances in various approaches and new tools used for cultivation of microalgae on IWW such as two-step cultivation, pre-acclimatization, novel microalgal-bioelectrical systems, integrated catalytic intense pulse-light process, sequencing batch reactor, use of old stabilized algal-bacterial consortium, immobilized microalgae cells, microalgal bacterial membrane photobioreactor, low-intensity magnetic field, BIO_ALGAE simulation tool, etc. are discussed. In addition, biorefinery of microalgal biomass grown on IWW and its end-use applications are reviewed.

Physiological responses of the green microalga Acutodesmus dimorphus to temperature induced oxidative stress conditions.

Temperature is the most critical factor that directly affects the physiological functioning and metabolic activities of any organism. With rising global temperature, understanding the heat stress response of an organism is critically important. In the present study, we investigated differences in the early changes occurring upon heat stress in the green microalga Acutodesmus dimorphus, a potential strain for biofuel production. The cells were heat-stressed at 45 and 50°C for 24 h and the temporal response of cells in terms of growth, pigments content, levels of oxidative stress biomarkers i.e., reactive oxygen species (ROS) and the response of enzymatic and non-enzymatic antioxidant scavengers were evaluated. The results revealed that after 24 h of heat stress at 45°C, the accumulations of chlorophyll a and carotenoids remained stable; all three ROS increased with the higher activities of various enzymatic and non-enzymatic antioxidants. On the contrary, at a higher temperature of 50°C, the accumulations of chlorophyll a, carotenoids and non-enzymatic antioxidants reduced drastically while the accumulations of all three ROS and the response of enzymatic antioxidants were significantly higher than those at 45°C. These results suggest that the cells utilize several stress acclimatization mechanisms to cope up the heat stress. There was a dramatic difference in the physiological changes and cellular antioxidant mechanism upon heat stress at 45 and 50°C. The cellular defense response of A. dimorphus gets impaired after heat stress at 50°C but remains active at 45°C, exhibiting the heat resistance and, thus, the thermotolerance.

Non-isothermal pyrolysis of de-oiled microalgal biomass: Kinetics and evolved gas analysis.

Non-isothermal (ß=5, 10, 20, 35°C/min) pyrolysis of de-oiled microalgal biomass (DMB) of Chlorella variabilis was investigated by TGA-MS (30-900°C, Argon atmosphere) to understand thermal decomposition and evolved gas analysis (EGA). The results showed that three-stage thermal decomposition and three volatilization zone (100-400°C, 400-550°C and 600-750°C) of organic matters during pyrolysis. The highest rate of weight-loss is 8.91%/min at 302°C for 35°C/min heating-rate. Kinetics of pyrolysis were investigated by iso-conversional (KAS, FWO) and model-fitting (Coats-Redfern) method. For Zone-1and3, similar activation energy (Ea) is found in between KAS (α=0.4), FWO (α=0.4) and Avrami-Erofe'ev (n=4) model. Using the best-fitted kinetic model Avrami-Erofe'ev (n=4), Ea values (R2=>0.96) are 171.12 (Zone-1), 404.65 (Zone-2) and 691.42kJ/mol (Zone-3). EGA indicate the abundance of most gases observed consequently between 200-300°C and 400-500°C. The pyrolysis of DMB involved multi-step reaction mechanisms for solid-state reactions having different Ea values.

Applications of de-oiled microalgal biomass towards development of sustainable biorefinery.

In view of commercialization of microalgal biofuel, the de-oiled microalgal biomass (DMB) is a surplus by-product in the biorefinery process that needs to be exploited to make the process economically attractive and feasible. This DMB, rich in carbohydrates, proteins, and minerals, can be used as feed, fertilizer, and substrate for the production of bioethanol/bio-methane. Further, thermo-chemical conversion of DMB results into fuels and industrially important chemicals. Future prospects of DMB also lie with its conversion into novel biomaterials like nanoparticles and carbon-dot which have biomedical importance. The lowest valued application of DMB is to use it for adsorption of dyes and heavy metals from industrial effluents. This study reviews how DMB can be utilized for different applications and in the generation of valuable co-products. The value addition of DMB would thereby improve the overall cost economics of the microalgal bio-refinery.

Hydrolysate of lipid extracted microalgal biomass residue: An algal growth promoter and enhancer.

The present study demonstrates the utilization of the algal hydrolysate (AH) prepared from lipid extracted residual harmful bloom-forming cyanobacteria Lyngbya majuscula biomass, as a growth supplement for the cultivation of green microalgae Chlorella vulgaris. BG-11 replacements with AH in different proportions significantly affects the cell count, dry cell weight (DCW), biomass productivity (BP) and pigments concentration. Among all, 25% AH substitution in BG11 media was found to be optimum which enhanced DCW, BP and pigments content by 39.13%, 40.81% and 129.47%, respectively, compared to control. The lipid content (31.95%) was also significantly higher in the 25% AH replacement. The volumetric productivity of neutral lipids (ideal for biodiesel) and total protein content of the cells significantly increased in all AH substitutions. Thus, lipid extracted microalgal biomass residue (LMBR) hydrolysate can be a potential growth stimulating supplement for oleaginous microalgae C. vulgaris.

Comparative evaluation of chemical and enzymatic saccharification of mixotrophically grown de-oiled microalgal biomass for reducing sugar production.

For the commercialization of microalgal based biofuels, utilization of de-oiled carbohydrate rich biomass is important. In the present study, chemo-enzymatic hydrolysis of mixotrophically grown Scenedesmus sp. CCNM 1077 de-oiled biomass is evaluated. Among the chemical hydrolysis, use of 0.5M HCl for 45 min at 121°C resulted in highest saccharification yield of 37.87% w/w of de-oiled biomass. However, enzymatic hydrolysis using Viscozyme L at loading rate of 20 FBGU/g of de-oiled biomass, pH 5.5 and temperature 45°C for 72 h resulted in saccharification yield of 43.44% w/w of de-oiled biomass. Further, 78% ethanol production efficiency was achieved with enzymatically hydrolyzed de-oiled biomass using yeast Saccharomyces cerevisiae ATCC 6793. These findings of the present study show application of mixotrophically grown de-oiled biomass of Scenedesmus sp. CCNM 1077 as promising feedstock for bioethanol production.

Selective carotenoid accumulation by varying nutrient media and salinity in Synechocystis sp. CCNM 2501.

Nutrients are the deciding factors in the biological production of bioactive compounds. Various growth media like BG11, Zarrouk's and Chu's 10 were studied for carotenoid production in Synechocystis sp. CCNM 2501. Maximum carotenoid content (dry weight basis) was found in Zarrouk's medium (ZM, 7.99mgg(-1)) followed by BG11 (5.13mgg(-1)). Echinenone content was 4 times higher in ZM (3.81mgg(-1)) as compared to BG11 (0.95mgg(-1)) and Chu's 10 (0.77mgg(-1)). Being an economical medium, BG11 was selected for carotenoid production. Further, increase in salinity from 0 to 0.2M in BG11 medium increases total carotenoid content from 5.82 to 7.05mgg(-1) and later it declines to 6.23mgg(-1) (1M). 3 times more ß-carotene is produced at 1M salinity as compared to control BG11. The variation in carotenoid composition with change in nutrients/salinity can be a good strategy to enhance certain targeted carotenoids.

Antioxidant, Anti-Nephrolithe Activities and in Vitro Digestibility Studies of Three Different Cyanobacterial Pigment Extracts.

Phycobiliprotein-containing water and carotenoid-containing methanolic extracts of three different cyanobacteria, Pseudanabaena sp., Spirulina sp. and Lyngbya sp., were studied for their DPPH scavenging, iso-bolographic studies, and anti-nephrolithe activities. The best EC50 values for DPPH scavenging were in Lyngbya water (LW, 18.78 ± 1.57 mg·mg(-1) DPPH) and Lyngbya methanol (LM, 59.56 ± 37.38 mg·mg(-1) DPPH) extracts. Iso-bolographic analysis revealed most of the combinations of extracts were antagonistic to each other, although LM-Spirulina methanol (SM) 1:1 had the highest synergistic rate of 86.65%. In vitro digestion studies showed that DPPH scavenging activity was considerably decreased in all extracts except for Pseudanabaena methanol (PM) and LM after the simulated digestion. All of the extracts were effective in reducing the calcium oxalate crystal size by nearly 60%-65% compared to negative control, while PM and Spirulina water (SW) extracts could inhibit both nucleation and aggregation of calcium oxalate by nearly 60%-80%.

Bicarbonate supplementation enhanced biofuel production potential as well as nutritional stress mitigation in the microalgae Scenedesmus sp. CCNM 1077.

The aim of the present study was to find out the optimum sodium bicarbonate concentration to produce higher biomass with higher lipid and carbohydrate contents in microalgae Scenedesmus sp. CCNM 1077. The role of bicarbonate supplementation under different nutritional starvation conditions was also evaluated. The results clearly indicate that 0.6 g/L sodium bicarbonate was optimum concentration resulting in 20.91% total lipid and 25.56% carbohydrate along with 23% increase in biomass production compared to normal growth condition. Addition of sodium bicarbonate increased the activity of nutrient assimilatory enzymes, biomass, lipid and carbohydrate contents under different nutritional starvation conditions. Nitrogen starvation with bicarbonate supplementation resulted in 54.03% carbohydrate and 34.44% total lipid content in microalgae Scenedesmus sp. CCNM 1077. These findings show application of bicarbonate grown microalgae Scenedesmus sp. CCNM 1077 as a promising feedstock for biodiesel and bioethanol production.

Salinity induced oxidative stress enhanced biofuel production potential of microalgae Scenedesmus sp. CCNM 1077.

Microalgal biomass is considered as potential feedstock for biofuel production. Enhancement of biomass, lipid and carbohydrate contents in microalgae is important for the commercialization of microalgal biofuels. In the present study, salinity stress induced physiological and biochemical changes in microalgae Scenedesmus sp. CCNM 1077 were studied. During single stage cultivation, 33.13% lipid and 35.91% carbohydrate content was found in 400 mM NaCl grown culture. During two stage cultivation, salinity stress of 400 mM for 3 days resulted in 24.77% lipid (containing 74.87% neutral lipid) along with higher biomass compared to single stage, making it an efficient strategy to enhance biofuel production potential of Scenedesmus sp. CCNM 1077. Apart from biochemical content, stress biomarkers like hydrogen peroxide, lipid peroxidation, ascorbate peroxidase, proline and mineral contents were also studied to understand the role of reactive oxygen species (ROS) mediated lipid accumulation in microalgae Scenedesmus sp. CCNM 1077.

Draft Genome Sequence of Halomonas hydrothermalis MTCC 5445, Isolated from the West Coast of India.

We announce here the draft genome sequence of Halomonas hydrothermalis MTCC 5445, a halophilic bacterium of the class Gammaproteobacteria. It was isolated from the sea coast of Aadri, Veraval, Gujarat, India. Its genome contains genes for polyhydroxybutyrate (PHB), a biodegradable polymer that can be used as a substitute for petroleum plastics.

Biofuel potential of the newly isolated microalgae Acutodesmus dimorphus under temperature induced oxidative stress conditions.

Lack of control over temperature is one of the major issues in large scale cultivation of microalgae. Therefore, it is important to evaluate the effects of cultivation temperature on the growth and physiology of microalgae. In the present study, freshwater microalgae Acutodesmus dimorphus was grown at different temperature in continuous and two stage cultivation. Results revealed that during continuous cultivation A. dimorphus grows better at 35°C than at 25°C and 38°C. At 35°C, A. dimorphus produced 22.7% lipid (containing 59% neutral lipid) and 33.7% carbohydrate along with 68% increase in biomass productivity (23.53mg/L/day) compared to 25°C grown culture. Stress biomarkers like reactive oxygen species, antioxidant enzymes like catalase and ascorbate peroxidase and lipid peroxidation were also lowest in 35°C grown culture which reveals that A. dimorphus is well acclimatized at 35°C.

Lipid Extracted Microalgal Biomass Residue as a Fertilizer Substitute for Zea mays L.

High volumes of lipid extracted microalgal biomass residues (LMBRs) are expected to be produced upon commencement of biodiesel production on a large scale, thus necessitating its value addition for sustainable development. LMBRs of Chlorella variabilis and Lyngbya majuscula were employed to substitute the nitrogen content of recommended rate of fertilizer (RRF) for Zea mays L. The pot experiment comprised of 10 treatments, i.e., T1 (No fertilizer); T2 (RRF-120 N: 60 P2O5: 40 K2O kg ha(-1)); T3 to T6-100, 75, 50, and 25% N through LMBR of the Chlorella sp., respectively; T7 to T10-100, 75, 50, and 25% N through LMBR of Lyngbya sp., respectively. It was found that all LMBR substitution treatments were at par to RRF with respect to grain yield production. T10 gave the highest grain yield (65.16 g plant(-1)), which was closely followed by that (63.48 g plant(-1)) under T5. T10 also recorded the highest phosphorus and potassium contents in grains. T4 was markedly superior over control in terms of dry matter accumulation (DMA) as well as carbohydrate content, which was ascribed to higher pigment content and photosynthetic activity in leaves. Even though considerably lower DMA was obtained in Lyngbya treatments, which might have been due to the presence of some toxic factors, no reduction in grain yield was apparent. The length of the tassel was significantly higher in either of the LMBRs at any substitution rates over RRF, except T6 and T7. The ascorbate peroxidase activity decreased with decreasing dose of Chlorella LMBR, while all the Lyngbya LMBR treatments recorded lower activity, which were at par with each other. Among the Chlorella treatments, only T5 recorded significantly higher values of glutathione reductase activity over RRF, while the rest were at par. There were significant increases in carbohydrate and crude fat, respectively, only in T4 and T3 over RRF, while no change was observed in crude protein due to LMBR treatments. Apparently, there was no detrimental effect on soil properties, suggesting that both the LMBRs can be employed to reduce the usage of chemical fertilizers, thus promoting maize crop production in a sustainable manner.

Biosorption of methylene blue by de-oiled algal biomass: equilibrium, kinetics and artificial neural network modelling.

The main objective of the present study is to effectively utilize the de-oiled algal biomass (DAB) to minimize the waste streams from algal biofuel by using it as an adsorbent. Methylene blue (MB) was used as a sorbate for evaluating the potential of DAB as a biosorbent. The DAB was characterized by SEM, FTIR, pHPZC, particle size, pore volume and pore diameter to understand the biosorption mechanism. The equilibrium studies were carried out by variation in different parameters, i.e., pH (2-9), temperature (293.16-323.16 K), biosorbent dosage (1-10 g L(-1)), contact time (0-1,440 min), agitation speed (0-150 rpm) and dye concentration (25-2,500 mg L(-1)). MB removal was greater than 90% in both acidic and basic pH. The optimum result of MB removal was found at 5-7 g L(-1) DAB concentration. DAB removes 86% dye in 5 minutes under static conditions and nearly 100% in 24 hours when agitated at 150 rpm. The highest adsorption capacity was found 139.11 mg g(-1) at 2,000 mg L(-1) initial MB concentration. The process attained equilibrium in 24 hours. It is an endothermic process whose spontaneity increases with temperature. MB biosorption by DAB follows pseudo-second order kinetics. Artificial neural network (ANN) model also validates the experimental dye removal efficiency (R2 = 0.97) corresponding with theoretically predicted values. Sensitivity analysis suggests that temperature and agitation speed affect the process most with 23.62% and 21.08% influence on MB biosorption, respectively. Dye adsorption capacity of DAB in fixed bed column was 107.57 mg g(-1) in preliminary study while it went up to 139.11 mg g(-1) in batch studies. The probable mechanism for biosorption in this study is chemisorptions via surface active charges in the initial phase followed by physical sorption by occupying pores of DAB.

Nitrogen stress triggered biochemical and morphological changes in the microalgae Scenedesmus sp. CCNM 1077.

The aim of present study was to investigate the effects of nitrogen limitation as well as sequential nitrogen starvation on morphological and biochemical changes in Scenedesmus sp. CCNM 1077. The results revealed that the nitrogen limitation and sequential nitrogen starvation conditions significantly decreases the photosynthetic activity as well as crude protein content in the organism, while dry cell weight and biomass productivity are largely unaffected up to nitrate concentration of about 30.87mg/L and 3 days nitrate limitation condition. Nitrate stress was found to have a significant effect on cell morphology of Scenedesmus sp. CCNM 1077. Total removal of nitrate from the growth medium resulted in highest lipid (27.93%) and carbohydrate content (45.74%), making it a potential feed stock for biodiesel and bio-ethanol production. This is a unique approach to understand morphological and biochemical changes in freshwater microalgae under nitrate limitation as well as sequential nitrate removal conditions.

Effect of light quality on the C-phycoerythrin production in marine cyanobacteria Pseudanabaena sp. isolated from Gujarat coast, India.

The isolated cyanobacterium containing biopigments like chlorophyll-a, phycoerythrin, phycocyanin, and carotenoid was cultured under different quality of light modes to ascertain biomass and pigment productivity. On the basis of 16S rRNA gene sequence, the isolate was identified as Pseudanabaena sp. Maximum biomass concentration obtained in white-, blue-, and green-light was 0.82, 0.94, and 0.89 g/L, respectively. It was observed that maximum phycoerythrin production was in green light (39.2 mg/L), ensued by blue light (32.2 mg/L), while phycocyanin production was maximum in red light (10.9 mg/L). In yellow light, pigment production as well as the growth rate gradually declined after 12 days. Carotenoid production decreased in blue-, white-, and red-light after 15 days, while in green light it had increased gradually. The present communication suggests that Pseudanabaena sp. can be used for commercial production of phycoerythrin when grown under green light.