Marine Chlorella Vulgaris Research Articles

Biodegradation of benzo[a]pyrene by a marine Chlorella vulgaris LH-1 with heterotrophic ability

In this study, a microalga, Chlorella vulgaris LH-1, with heterotrophic ability to degrade BaP was explored. The effect of BaP concentration on microalga growth was investigated, and the possible biodegradation mechanism of BaP was proposed. Results showed that low BaP concentration (<5 mg/L) had less negative influence on the growth of this microalga under mixotrophic condition, but high BaP concentration (>5 mg/L) had a significant inhibitory effect on its growth. During heterotrophic cultivation, low BaP concentration (<20 mg/L) promoted the growth of C. vulgaris LH-1, whereas high BaP concentration (>20 mg/L) inhibited its growth significantly. The degradation rates of mixotrophic and heterotrophic C. vulgaris LH-1 were 62.56 %–74.13 % and 52.07 %–71.67 %, respectively, when the BaP concentration ranged from 0.5 mg/L to 2 mg/L. The expression of functional enzyme genes of C. vulgaris LH-1 such as phenol 2-monooxygenase activity, protocatechuate 3,4-dioxygenase activity, catechol 1,2-dioxygenase activity, styrene degradation, and benzoate degradation were upregulated in the process of BaP degradation. C. vulgaris LH-1 may degrade BaP by monooxygenase and dioxygenase simultaneously. The degradation of BaP by this microalga under mixotrophic condition goes through the degradation pathway of phthalic acid, whereas it goes through the degradation pathway of benzoic acid under heterotrophic condition.

RNA-sequencing analysis reveals the co-biodegradation performance of crude oil by marine Chlorella vulgaris under norfloxacin stress

In this study, the microalgal growth and crude oil (CRO) biodegradation by marine Chlorella vulgaris (C. vulgaris) were assessed under norfloxacin (NFX) stress. The presence of NFX negatively affected the bio-removal of CRO within 5 days, as the NFX concentration increased from 100 to 1600 μg/L, due to its toxicity as an antibiotic. However, its negative impact on the final degradation capabilities of C. vulgaris was less significant (P-value <0.05). After 9 days of cultivation, CRO bio-removal efficiencies still exceeded 90 %, while NFX bio-removal efficiencies maintained over 47 %. RNA-seq analysis revealed that the degradation of CRO and NFX was attributed to the combined action of functional genes involved in scavenging reactive oxygen species. The production of pigments and the bio-removal performance of C. vulgaris in CRO, NFX, and CRO & NFX coexistence media were consistent with the changes in the number of differentially expressed genes in these samples.

Design and fabrication of box-type passive solar dryer (BTPSD) with thermal insulation material for valorizing biomass and neutral lipids of marine Chlorella vulgaris for biodiesel application

The staggering rate of population growth has augmented the reliance on fossil fuel utilization, and it kindled the society to explore alternative and sustainable sources of energy. In this regard, biodiesel from microalgae came to the limelight; but crucial energy-consuming and expensive processes like cultivation, harvesting, and drying make the microalgal biodiesel unsustainable and economically unfeasible. To surpass these impediments, in this research work, a low-cost box-type passive solar dryer (BTPSD) is designed and fabricated with zero energy consumption mode and compared with conventional hot air oven for drying the biomass, neutral lipids of the marine microalga Chlorella vulgaris for biodiesel application. The onset of the work, BTPSD with 2 cm thickness of glass wool as TIM (thermal insulation material), 4 cm TIM thickness and no TIM was simulated for thermal storage behaviour using ANSYS FLUENT 19.2 Computational Fluid Dynamics tool and based on the results, 4 cm TIM thickness was chosen for experimentation. The time taken by BTPSD and hot air oven to remove the moisture from algal biomass is 3 and 2 h, respectively, whereas for neutral lipids drying, it was 4 and 3.5 h, respectively. Though there is a little difference in drying time, neutral lipid and FAME content from both drying systems are tantamount, i.e., ~ 12% neutral lipid and 95% FAME. Further, the percentage of vital fatty acids identified from BTPSD and hot air oven methods are almost similar, i.e., C16:0 (23.4%), C18:1 (14.3%), C18:3 (11.42%), C18:1 (9.22%). Though the time taken for valorizing biomass and neutral lipids of C. vulgaris by BTPSD is slightly longer than hot air oven, low energy consumption and cost-effectiveness make the BTPSD a promising system to scale down the microalgal biodiesel production cost significantly.

A Single Dose of Marine Chlorella vulgaris Increases Plasma Concentrations of Lutein, β-Carotene and Zeaxanthin in Healthy Male Volunteers.

The beneficial health effects of Chlorella vulgaris have been associated with the presence of several nutrients and antioxidants, including carotenoids. However, the in vivo bioavailability of Chlorella is still poorly evaluated. In this work, a human intervention study was conducted in 11 healthy men to evaluate the bioavailability of carotenoids within 3 days after the intake of a single dose (6 g) of dried marine Chlorella vulgaris containing lutein (7.08 mg), β-carotene (1.88 mg) and zeaxanthin (1.47 mg). Subjects were instructed to follow a low carotenoid diet during the experimental phase, starting 1 week earlier. On the day of the experiment, dried microalgae formulated in vegetarian hard capsules were ingested, and blood samples were collected up to 72 h for the analysis of plasma carotenoids concentration by high-performance liquid chromatography with diode-array detection. For all carotenoids, the estimated AUC and Cmax values were significantly different from zero (p < 0.05), indicating that a single dose of marine Chlorella vulgaris increased plasma concentrations of lutein (Cmin-corrected AUC = 1002 µg·h/L, Cmax = 20.4 µg/L), β-carotene (AUC = 1302 µg·h/L, Cmax = 34.9 µg/L) and zeaxanthin (AUC = 122.2 µg·h/L, Cmax = 3.4 µg/L). The bioavailability of other compounds, namely, polyunsaturated fatty acids and trace elements, was also assessed post-prandial for the first time, showing that linoleic acid, docosahexaenoic acid and iodine were absorbed after microalgae intake. These findings support the use of Chlorella vulgaris as a source of carotenoids, PUFA and essential trace elements with associated health benefits.

Novel High-Selectivity Fluorescent Probe for Detecting Alkaline Phosphatase Activity in Marine Environment.

In marine environment, alkaline phosphatase (ALP) is a nonspecific phosphatase with metal ions as its active site. Metal ions have different effects on ALP activity. Therefore, a probe that specifically detects ALP needs to be developed. In this paper, to eliminate the interference of acid phosphatase, we designed and synthesized a highly selective fluorescent probe CyP based on pH to detect ALP activity. The response mechanism of detecting ALP was explained. The photophysical properties, enzyme kinetics, stability, selectivity, and potential quantitative ability of the probe under different pH values were investigated. The effects of metal ions on the ALP activity of marine Chlorella vulgaris and Escherichia coli were also analyzed. Excessive metal ions such as Zn2+, Cu2+, Hg2+, Pb2+, and Cd2+ inhibit while Mn2+, Co2+, and alkaline-earth metal ions promote the ALP activity of Chlorella and E. coli.

Simultaneous removal of nutrient and sulfonamides from marine aquaculture wastewater by concentrated and attached cultivation of Chlorella vulgaris in an algal biofilm membrane photobioreactor (BF-MPBR)

Microalgae based wastewater treatment has attracted increasing attention for its many advantages in recent years. In this study, a novel microalgae biofilm membrane photobioreactor (BF-MPBR) was developed for the efficient microalgae cultivation and the removal of nutrient and sulfonamides (SAs) from marine aquaculture wastewater. Two BF-MPBRs with hydraulic retention time (HRT) of 1 day and 2 days respectively were continuously operated for 70 days without harvesting microalgae. Concentrated and attached culture of marine Chlorella vulgaris was achieved in these continuous flow BF-MPBRs due to the suspended solid carriers and microfiltration membrane module in the reactors. The algal biomass productivity achieved in BF-MPBRs with HRT of 1 day and 2 days were 14.02 and 22.03 mg L−1 day−1, respectively. In addition, at the end of the cultivation, 60.4% and 45.0% of microalgae were fixed into algal biofilm in BF-MPBRs with 1 day and 2 day HRT, respectively. Compared with batch cultivation, more efficient nutrient and SAs removal performance was achieved in BF-MPBRs, although the HRT of the BF-MPBRs used in this study was only 1 or 2 days. During the stable operation stage of the BF-MPBRs, the reduction in dissolved inorganic nitrogen (DIN), dissolved inorganic phosphorus (DIP), sulfadiazine (SDZ), sulfamethazine (SMZ) and sulfamethoxazole (SMX) were found in the range of 91.0–99.6%, 92.1–98.4%, 61.0–79.2%, 50.0–76.7% and 60.8–82.1%, respectively. Therefore, nutrient and SAs were simultaneously and efficiently removed from marine aquaculture wastewater by microalgae cultivation in BF-MPBR.

RNA-seq analysis reveals the significant effects of different light conditions on oil degradation by marine Chlorella vulgaris

Marine Chlorella vulgaris, an efficient hydrocarbon-degrading organism, is easily affected by light. In this study, we investigated the direct effects of different light conditions on crude oil degradation by C. vulgaris and its crude enzyme. Under 12 h illumination, the crude enzyme improved hydrocarbon removal by 39.36%, whereas the addition of the enzyme and C. vulgaris increased the degradation rate by 121.73%. Conversely, the addition of enzyme under heterotrophic condition was negatively related to oil degradation by C. vulgaris, and the degradation rate decreased from 74.32% to 48.65% and further reduced by 34.54%. The results of RNA sequencing analysis suggested that hydrocarbons removal was attributed to C. vulgaris metabolism in heterotrophic physiological state. While enhanced removal efficiency of hydrocarbons was achieved in mixotrophic physiological state due to the coupling of C. vulgaris metabolism with photocatalytic oxidation. Functional enzymes played key roles in photocatalysis and biodegradation processes.

Growth characteristics of shade-acclimated marine Chlorella vulgaris under high-cell-density conditions

Growth characteristics of shade-acclimated marine Chlorella vulgaris under high-cell-density conditions

Semicontinuous outdoor cultivation and efficient harvesting of marine Chlorella vulgaris BDUG 91771 with minimum solid co-precipitation and high floc recovery for biodiesel

The present study demonstrated contamination free, semicontinuous mode cultivation of marine Chlorella vulgaris BDUG 91771 at 5kL open pond. Culture cultivated in seawater supplemented with inexpensive urea and superphosphate exhibited specific growth rate, biomass productivity, lipid content, lipid productivity, and doubling time of 0.703d−1, 0.056gL−1d−1, 20%, 0.012gL−1d−1 and 37–40h respectively at 30°C average system temperature and 30o/00 salinity. With an aim to develop an appropriate harvesting technique based on maximal floc recovery and less solid co-precipitation, various chemical, natural, electroflocculation methods were investigated. Of the flocculants, 70mgL−1 aluminium sulfate, 20mgL−1 polymer flocculant, Cu electrode mediated electroflocculation showed ∼98% floc recovery at 120min flocculation time whereas natural flocculant Moringa seed powder even at a high dose of 700mgL−1 divulged low floc recovery (63.06%). Notably, polymer flocculant exhibited the minimum co-precipitation of solid and high biomass content of 13mgL−1 and 147mgL−1 respectively compared to other flocculants, and it also had stable, intact viable cells in flocs reflected through dye exclusion staining. This study further stated that the critical elements namely Ca2+ (1.4mM), and Mg2+ (0.27mM) in the marine medium are not required for polymer based flocculation and were essential for chemical based flocculation. Eventually, the fatty acid pool of laboratory and outdoor grown biomass reflected the same pattern and possess C16:0 C16:1, C18:1 for high-quality biodiesel.

Efficient harvesting of marine Chlorella vulgaris microalgae utilizing cationic starch nanoparticles by response surface methodology

Harvesting involves nearly thirty percent of total production cost of microalgae that needs to be done efficiently. Utilizing inexpensive and highly available biopolymer-based flocculants can be a solution for reducing the harvest costs. Herein, flocculation process of Chlorella vulgaris microalgae using cationic starch nanoparticles (CSNPs) was evaluated and optimized through the response surface methodology (RSM). pH, microalgae and CSNPs concentrations were considered as the main independent variables. Under the optimum conditions of microalgae concentration 0.75gdry weight/L, CSNPs concentration 7.1mgdry weight/L and pH 11.8, the maximum flocculation efficiency (90%) achieved. Twenty percent increase in flocculation efficiency observed with the use of CSNPs instead of the non-particulate starch which can be due to the more electrostatic interactions between the cationic nanoparticles and the microalgae. Therefore, the synthesized CSNPs can be employed as a convenient and economical flocculants for efficient harvest of Chlorella vulgaris microalgae at large scale.

Assessment of fuel properties, engine performance and emission characteristics of outdoor grown marine Chlorella vulgaris BDUG 91771 biodiesel

The present study investigates the suitability of marine microalga Chlorella vulgaris BDUG 91771 biodiesel to curb the fuel crisis, based on engine performance and emission profile. Initially, biodiesel was produced from outdoor grown Chlorella vulgaris BDUG 91771 and its fuel properties accorded with ASTM D6571 and EN 14214 standards. To evaluate performance and emission characteristics, various Chlorella vulgaris BDUG 91771 biodiesel-diesel blends were prepared and tested in single-cylinder Kirloskar engine under four different load operations (0%, 50%, 75%, 100%). Compression ignition engine fuelled with B40 or B50 biodiesel-diesel blend showed specific fuel consumption (SFC), and brake thermal efficiency (BTE) of 0.306 kg/kWh and 26% respectively at 100% load, which is on par with diesel fuel. Further, engine operated with B50 biodiesel blend evidently reduced the carbon monoxide (CO) and hydrocarbon (HC) emissions by 0.1% and 102 ppm respectively compared to baseline diesel. Notably, diesel emitted 7.5% carbon dioxide (CO2) and 380 ppm nitrogen oxides (NOx) at 100% load, while the optimal blends B40 or B50 reduced CO2 and NOx emissions by 6.1% and 376 ppm respectively. Based on results, present work suggests either B40 or B50 Chlorella vulgaris BDUG 91771 biodiesel – diesel blend could serve as eco-friendly, alternative fuel.

Harvesting of microalgae by flocculation with poly (γ-glutamic acid)

In an effort to search for an efficient and environmentally friendly harvesting method, a commercially available microbial flocculant poly (γ-glutamic acid) (γ-PGA) was used to harvest oleaginous microalgae. Conditions for flocculation of marine Chlorella vulgaris and freshwater Chlorella protothecoides were optimized by response surface methodology (RSM) and determined to be 22.03mgL−1 γ-PGA, 0.57gL−1 biomass, and 11.56gL−1 salinity, and 19.82mgL−1 γ-PGA and 0.60gL−1 biomass, respectively. Application of the two optimized flocculation methods to Nannochloropsis oculata LICME 002, Phaeodactylum tricornutum, C. vulgaris LICME 001, and Botryococcus braunii LICME 003 gave no less than 90% flocculation efficiency and a concentration factor greater than 20. Micrographs of the harvested microalgal cells showed no damage to cell integrity, and hence no lipid loss during the process. The results show that flocculation with γ-PGA is feasible for harvesting microalgae for biodiesel production.

Disruption of Chlorella vulgaris Cells for the Release of Biodiesel-Producing Lipids: A Comparison of Grinding, Ultrasonication, Bead Milling, Enzymatic Lysis, and Microwaves

A comparative evaluation of different cell disruption methods for the release of lipids from marine Chlorella vulgaris cells was investigated. The cell growth of C. vulgaris was observed. Lipid concentrations from different disruption methods were determined, and the fatty acid composition of the extracted lipids was analyzed. The results showed that average productivity of C. vulgaris biomass was 208 mg L⁻¹ day⁻¹. The lipid concentrations of C. vulgaris were 5%, 6%, 29%, 15%, 10%, 7%, 22%, 24%, and 18% when using grinding with quartz sand under wet condition, grinding with quartz sand under dehydrated condition, grinding in liquid nitrogen, ultrasonication, bead milling, enzymatic lysis by snailase, enzymatic lysis by lysozyme, enzymatic lysis by cellulose, and microwaves, respectively. The shortest disruption time was 2 min by grinding in liquid nitrogen. The unsaturated and saturated fatty acid contents of C. vulgaris were 71.76% and 28.24%, respectively. The extracted lipids displayed a suitable fatty acid profile for biodiesel [C16:0 (~23%), C16:1 (~23%), and C18:1 (~45%)]. Overall, grinding in liquid nitrogen was identified as the most effective method in terms of disruption efficiency and time.

The toxicity of naphthalene to marine Chlorella vulgaris under different nutrient conditions

The toxicity of naphthalene to Chlorella vulgaris was studied under nitrogen (N)-, phosphorus (P)-enriched and N,P-starved condition. Results showed that naphthalene was less toxic under N,P-starved condition. The inhibitory rates were less than 15.3% to C. vulgaris during 7 days exposure with the initial concentrations of naphthalene at 5, 10, 50, 100mg/L, respectively under N,P-starved condition, while they were 7.5-72.3% under N,P-enriched condition. The malondialdehyde (MDA) content indicated that there was no oxidative damage to algae when the initial concentration of naphthalene was less than 10mg/L, and oxidative damage exhibited to algae at 50-100mg/L of naphthalene under N,P-starved condition. Naphthalene induced oxidative damage to the algae at all tested concentrations (5-100mg/L) under N,P-enriched condition. The results indicated that there was a negative relationship between the special growth rate (SGR) and naphthalene concentration in the medium. Under N,P-enriched condition SGR of the control decreased slowly from 0.669 to 0.186. However, SGR of the naphthalene treated group decreased sharply during the first 2-3 days when the dissolved concentration of naphthalene was above 0.1mg/L, and then increased gradually with the evaporation of naphthalene.

Enhancement of adhesion of the marine Chlorella vulgaris to glass.

The adhesion of washed cells of a marine Chlorella vulgaris to solid surfaces was enhanced by non-diffusible material recovered from Chlorella exudate, marine bacterial cultures, natural seawater, and fouled marine surfaces. Materials isolated from certain bacterial cultures and from particulate materials filtered from seawater were three orders of magnitude more active than Chlorella exudate per unit weight. Active polymer materials from several sources were chromatographed on DEAE cellulose. The major fraction eluted with dilute base contained both protein and carbohydrate and enhanced adhesion more than the unchromatographed material.