Renewable biomass production by mixotrophic algae in the presence of various carbon sources and wastewaters

This study evaluated mixotrophic growth potential of native microalgae in media supplemented with different organic carbon substrates and wastewaters. Three robust mixotrophic microalgae viz. Chlamydomonas globosa, Chlorella minutissima and Scenedesmus bijuga were isolated after long-term enrichments from industrial wastewater. The mixotrophic growth of these microalgae resulted in 3–10 times more biomass production relative to phototrophy. Glucose, sucrose and acetate supported significant mixotrophic growth. Poultry litter extract (PLE) as growth medium recorded up to 180% more biomass growth compared to standard growth medium BG11, while treated and untreated carpet industry wastewaters also supported higher biomass, compared to BG11 growth with no significant effect of additional nitrogen supplementation. Supplementing treated wastewater and PLE with glucose and nitrogen resulted in 2–7 times increase in biomass relative to the unamended wastewaters or PLE. The consortia of Chlamydomonas–Chlorella and Scenedesmus–Chlorella were the best for PLE and untreated wastewater respectively, while a combination all three strains was suitable for both PLE and wastewater. These algae can be good candidates for biofuel feedstock generation as they would not require freshwater or fertilizers. Such mixotrophic algal consortia offer great promise for production of renewable biomass for bioenergy applications using wastewaters.     

Biomass and lipid production of marine microalgae using municipal wastewater and high concentration of CO2

In order to reduce the cost of the production of microalgae for biodiesel, the feasibility of using the mixture of seawater and municipal wastewater as culture medium and CO₂ from flue gas for the cultivation of marine microalgae was investigated in this study. Effects of different ratios of municipal wastewater and 15% CO₂ aeration on the growth of Nannochloropsis sp. were examined, and lipid accumulation of microalgae was also studied under nitrogen starvation and high light. It was found that optimal growth of microalgae occurred in 50% municipal wastewater, and the growth was further significantly enhanced by aeration with 15% CO₂. When Nannochloropsis sp. cells were transferred from the first growth phase to the second lipid accumulation phase under the combination of nitrogen deprivation and high light, both biomass and lipid production of Nannochloropsis sp. were significantly increased. After 12 days of the second-phase cultivation, the biomass concentration and total lipid content increased from 0.71 to 2.23 g L⁻¹ and 33.8–59.9%, respectively. This study suggests that it is possible to utilize municipal wastewater to replace nutrients in seawater medium and use flue gas to provide CO₂ in the cultivation of oil-bearing marine microalgae for biodiesel.     

Culture of four microalgal strains for bioenergy production and nutrient removal in the meliorative municipal wastewater

Microalgae have been considered as the most promising sources of alternative bioenergy. For the purpose of saving costs, the present work focused on the potential use of microalgae in the meliorative municipal wastewater, which contains 90% municipal wastewater and 10% dairy wash wastewater. Four microalgal species, Palmellococcus miniatus, Neochloris oleoabundans, Scenedesmus quadricanda #507, and Chlorella zofingiensis, were cultured in pure municipal wastewater and meliorative municipal wastewater, respectively, for 5 days. Their biomass accumulation and removal rates of nitrogen and phosphate were measured. Results showed that the growth rates of Neochloris oleoabundans, Palmellococcus miniatus, and Chlorella zofingiensis in meliorative municipal wastewater (>0.8 g·L^–1·d^–1) were significantly higher than that in municipal wastewater (2.6 g·L^–1·d^–1), while there was no significant difference between the growth rates of Scenedesmus quadricanda #507 in meliorative municipal wastewater and in municipal wastewater. Neochloris oleoabundans exhibited the highest growth rate (0.86 g·L^–1·d^–1) and relatively high nutrient removal capacity. Scenedesmus quadricanda #507 had the highest P removal rate of over 94%. The four species have a similar N removal rate at about 90%. The results showed that the highest average removal rate of N and P were about 23.1 mg·L^–1·d^–1 and 7.1 mg·L^–1·d^–1. Furthermore, the content of lipid or carbohydrates increased and a different profile of fatty acids were found compared to those in municipal wastewater. Cellular components changes of microalgae in meliorative municipal wastewater were favorable as raw materials for bioethanol and biodiesel production. Cultivation with meliorative municipal wastewater is a win-win culture mode that facilitates the biomass production, lipid and carbohydrate accumulation, and wastewater purification.     

Stress enhances poly-unsaturation rich lipid accumulation in Chlorella sp. and Chlamydomonas sp.

In the present study, effects of various cultivation conditions on biomass and lipid productivity, detail FAME signature, CO₂ biofixation and elemental composition of Chlorella sp. and Chlamydomonas sp. have been investigated. In Chlorella sp., N-depletion has enhanced the lipid productivity by 12.70–14.61%. In Chlamydomonas sp., though the lipid content has increased by 22–26% upon N-depletion, lipid productivity has not changed significantly due to concomitant decrease in biomass productivity by 18–25%. The presence of lipid was also confirmed by FTIR spectroscopic analysis of biomass. The characteristic bands observed at 1744.30 cm⁻¹ (Chlamydomonas sp.) and (Chlorella sp.) were credited to lipids due to C–O mode of the side chain from ester carbonyl group. Extracted lipid was transesterified to FAMEs. Chlorella sp. at N-replete indoor condition has produced the highest weight percentage of C16:0 (26.64%), C18:0 (3.77%) and C18:1 (27.94%). In Chlamydomonas sp., N-replete indoor condition has resulted in maximum content of MUFA (30.12%), LUFA (53.35%), second highest SFA (29.22%) and DUFA (23.23%). Inverse relationship was observed between MUFA and PUFA at all cultivation conditions. In both the microalgae, C16:0, C18:1 (9), C18:2 (9, 12), C18:3 (9, 12, 15) were identified as the major FAMEs which are suitable to be used as biodiesel components.     

Biomass and lipid productivities of marine microalgae isolated from the Persian Gulf and the Qeshm Island

In this work, the screening of 147 microalgal strains from the Persian Gulf and the Qeshm Island (Iran) were done in order to choose the best ones, in terms of growth (biomass) rate and lipid content for biodiesel production. A methodology, combining experiments in lab-scale and pilot plant (open pond) used to produce and evaluate biomass and lipid productivity is presented for the systematic investigation of the potential of different microalgae species. The culture conditions, including photo flux (180 ??E m⁻² s⁻¹), photoperiod (12 h light/dark), temperature (25 °C), pH (≈8), air (carbon dioxide) and growth medium, were kept constant for all experiments. Microalgae were screened in two stages using optical density (for evaluation of biomass concentration) and Nile red and gas chromatography (for determination of lipid content and fatty acid fractions). In general, maximum specific growth rate and the maximum biomass productivity were obtained after 8-12-day culture. Nannochloropsis sp. and Neochloris sp. were selected from the marine microalgal culture collection, due to their high biomass (50 and 21.7 g L⁻¹, respectively) and oil content (52% and 46%, respectively). If the purpose is to produce biodiesel only from one species, Nannochloropsis sp. presented the most adequate fatty acid profile, namely linolenic and other polyunsaturated fatty acids. However, the microalgae Chlorella sp. can also be used if associated with other microalgal oils. In addition, selected strains could be potent candidates for commercial production in the open pond culture.     

以市政污水为底物的微藻油脂积累和碳流分析

微藻培养的高成本阻碍了微藻生物柴油的工业化推广。文章以市政污水作为Chlorella vulgaris的培养基,分别在水力停留时间(HRT)24,48,96 h和间歇运行时对其进行培养。随着HRT的增加,C.vulgaris在系统生物群落中所占的比例增大,而且细胞密度和脂含量也分别由HRT 24 h时的0.088 g/L和8.4%提高到间歇时的0.164 g/L和14.3%。HRT对废水处理效果的影响较小,不同HRT时,COD和TP的去除率分别在66.1%~71.1%和37.6%~51.1%,TN的去除率则仅为5.9%~11.2%。对间歇污水处理过程碳元素流动分析表明,污水中的71.1%有机碳被微藻利用,其中94.6%被转化为微藻生物质,其余被代谢为CO2释放,废水中有机碳(COD)转化为微藻油脂的转化率为11.6 mg/(mg.L)。     

Coupling algal biomass production and anaerobic digestion: Production assessment of some native temperate and tropical microalgae

Coupling algal biomass production and anaerobic digestion is one of the most promising bioprocesses for economically viable algal production. This study assesses the production rates of some native microalgae growing in media supplemented with algal digestate, urban wastewater or digested sludge. Native microalgal populations isolated from temperate freshwaters (Scenedesmus spp.) and marine ecosystems (Nannochloris spp.) had the highest potential production rates (about 100 mg DW L⁻¹ d⁻¹) with algal digestate at about 20% loading ratio. However, no growth was measured for Nannochloris spp., when the ammonium concentration exceeded 100 mg L⁻¹ although Scenedesmus spp. appeared to be tolerant to higher NH₄⁺ concentrations. Very low production rates, or no growth, were measured when microalgae isolated from high salinity waters (Dunaliella salina, Lyngbya aestuarii) were used, suggesting that populations well adapted to extreme environmental conditions are not suitable candidates for growing on wastewater or anaerobic digestate.     

Culture of microalgal strains isolated from natural habitats in Thailand in various enriched media

Six freshwater microalgal strains in the class of Chlorophyceae, including Chlorococcum humicola, Didymocystis bicellularis, Monoraphidium contortum, Oocystis parva, Sphaerocystis sp., and Scenedesmus acutus were isolated from natural habitats in Thailand. The six strains were compared for their biomass yield, lipid content, and lipid productivity in four enriched culture media in batch mode. Significant differences were found across algal strains and culture media. The best strain was found to be C. humicola, which had the highest biomass yield of 0.113 g/l/d (in Kuhl medium), the highest lipid content of 45.94% (in BG-11 medium), and the highest lipid yield of 0.033 g/l/d (in 3NBBM medium). The 3NBBM medium, which has the lowest nitrogen concentration among the four culture media, was considered the optimal culture medium for C. humicola for lipid production. The fatty acid profile of C. humicola was also found to be affected by the culture medium. More oleic acid (C18:1) but less linolenic acid (C18:3) was accumulated in BG-11 and 3NBBM than in Kuhl and N-8 media. Lipid profiles of C. humicola were comparable to palm oil in the percentage of palmitic acid and the total amount of saturated fatty acids; however, C. humicola made more poly-unsaturated fatty acids such as linoleic (C18:2) and linolenic (C18:3) acids than oil palms. Lipids from C. humicola were believed to be acceptable for biodiesel production.     

Screening of marine microalgae for biodiesel feedstock

Biodiesel production from microalgae lipids is increasingly regarded as a more sustainable and feasible alternative to conventional biodiesel feedstocks derived from terrestrial bioenergy crops. A total of ninety-six strains of marine microalgae, with an elevated biomass productivity and intracellular lipid content, were isolated from the coastal waters of Singapore using an automated flow cytometric cell-sorting technique. Cell sorting was based on the two-dimensional distribution of algal cells for red fluorescence (representing chlorophyll auto-fluorescence) against forward-light scatter (representing cell size) and red vs. green fluorescence. Twenty-one of the strains were further characterized with respect to cell growth rate, biomass concentration, lipid content (total and neutral lipid) and fatty acid profile. The growth rates of Skeletonema costatum, Chaetoceros and Thalassiosira species were greatest among the entire strains, but in terms of absolute lipid yield Nannochloropsis strains predominated. Nannochloropsis strains had a lipid content ranging from 39.4% to 44.9% of dry weight biomass. Transesterification of the lipids yielded 25-51% of fatty acid methyl ester (FAME) i.e. biodiesel, where total FAME content ranged between 11 and 21% of dry weight biomass. This study describes the microalgae screening process and demonstrates that Nannochloropsis is a promising species for biodiesel feedstock.     

Lipid production in mixotrophic cultivation of Chlorella vulgaris in a mixture of primary and secondary municipal wastewater

Microalgae's biomass productivity and oil content depend heavily on the method of its cultivation. In this study, nutrient removal from municipal wastewater by Chlorella vulgaris in batch culture was investigated. Carbon dioxide was supplied from sodium hydrogen carbonate. Effect of parameters including light intensity, sodium hydrogen carbonate concentration, and daily illumination time on the productivity of biomass and lipid was investigated. Lipid and biomass production of C. vulgaris increased at higher concentration of sodium hydrogen carbonate concentration and higher light intensity until a certain value and then decreased, but longer daily illumination time, increased both biomass and lipid productivity. Cultivation of C. vulgairs in mixotrophic mode was also studied in a mixture of primary and secondary wastewater with different ratios (25, 50 and 75 volume percent of the primary wastewater). It was observed that using 25% of the primary wastewater results in 100% COD removal, 100% ammonium removal and 82% nitrate elimination. Biomass productivity and lipid productivity of C. vulgaris in a mixture of primary (25%) and secondary wastewater were 138.76 mg/L/d and 45.49 mg/L/d, respectively.     

Repeated batch fermentation for photo-hydrogen and lipid production from wastewater of a sugar manufacturing plant

Hydrogen and lipid production from sugar manufacturing plant wastewater (SMW) by Rhodobacter sp. KKU-PS1 were investigated. Aji-L (i.e., a waste from the process of crystallizing monosodium glutamate) was used as nitrogen source. Batch fermentation was conducted in 300 mL serum bottles with the working volume of 180 mL to investigate the optimal inoculum size by varying the initial inoculum concentration from 0.23 to 0.92 g_CDW/L. The photo-fermentation was conducted at an initial pH 7.0 and 25.6 °C with continuously light illumination at 7500 lux. The optimal inoculum size of 0.77 g_CDW/L gave the hydrogen production rate (R_m) and lipid production of 5.24 mL H₂/L.h and 407 mg lipid/L, respectively. The hydrogen production from SMW was further examined in 1.7-L photo-bioreactor with the working volume of 1.2-L using the optimal condition from batch experiment. A photo-bioreactor yielded 1.73 times higher R_m than that obtained from the fermentation in serum bottles with a greater lipid production of 424 mg lipid/L. Hydrogen production from SMW in the repeated-batch fermentation was further studied by varying the medium replacement ratios of 25, 50–75%. A maximum biomass and lipid concentration of 2.83 g_CDW/L and 685 mg lipid/L, respectively were achieved at a medium replacement ratio of 75%. C18:1 (51.2%), C18:0 (24.9%) and C16:0 (9.1%) were found as the major free fatty acid. Lactic acid followed by propionic, acetic and butyric acids containing in SMW were the suitable carbon source for biomass production of KKU-PS1.     

Evaluation of ultrasonication as a treatment strategy for enhancement of biohydrogen production from complex distillery wastewater and process optimization

Dark fermentation of distillery wastewater (DWW) gives a lower hydrogen yield (HY) and hydrogen production rate (R), owing to the complexity and a recalcitrant nature of effluent. Therefore, an effective pretreatment of DWW becomes imperative for the improvement of biohydrogen production. In the present study, the efficacy of ultrasonic pretreatment for enhancement of biohydrogen production from DWW was evaluated in batch test. Several variables, such as COD input, ultrasonic density (UD), and ultrasonication time (UT) were studied for optimization using response surface methodology integrated with desirability function. The highest HY, 10.95 mmol/g COD, and R, 6.67 mmol/L h, were obtained for batch test of ultrasonically pretreated DWW under optimal conditions for COD, UD and UT at 56 g/L, 0.12 W/mL, and 17 min, respectively. The significant relative enhancement of HY, 101%, and R, 103%, implies that ultrasonically treated DWW is about 1.2–1.4 times more effective for enhanced biohydrogen production from complex DWW compared to unsonicated DWW.     

Microbial oil production from thermophile cyanobacteria for biodiesel production

Compared to lipid extraction from algae, little work has been performed for cyanobacteria. In this article it is aimed to show high lipid accumulation potential of Synechococcus sp., Cyanobacterium aponinum and Phormidium sp. cells in BG-11 medium. Four different pH values (6–9) and NaNO₃ (0.25, 0.5, 1.0, 1.5 g/L) concentrations were examined at different incubation days to discover the highest lipid accumulation. The maximum lipid content could be achieved in the medium containing 0.25 g/L NaNO₃ at pH 7 for Synechococcus sp., pH 8 for C. aponinum and pH 9 for Phormidium sp. after 15 days. The maximum lipid contents and C16 and C18 methyl ester yields were measured as 42.8% and 46.9% for Synechococcus sp., 45.0% and 67.7% for C. aponinum, 38.2% and 90.6% for Phormidium sp. The saturated compounds were 74.5%, 77.9%, 84.7% for Synechococcus sp., C. aponinum and Phormidium sp., respectively. These crude lipids could be promising feedstock for biodiesel production.     

Enhancement of lipid productivity in green microalgae Chlorella sp. via fast neutron irradiation

Microalgae is becoming a promising candidate for biofuel production and energetic lipid production, which urges the need to develop efficient methods for improvement of the lipid production. In this study, the neutron irradiation was employed to improve lipid production under different dosage irradiation. Effects of different dosages (No.1:8.14 × 10⁹, No.2:5.64 × 10⁸, No.4:9.42 × 10⁷ n/cm²) were compared by investigating associated algae cell growth, total lipid production, metabolization and enzyme activity. Dosages No.2 and No.4 improved the lipid content without negative influence on the growing, whereas irradiations shortened the period of biomass accumulation. The neutron irradiation in No.2 accelerated the synthesis of lipid from the carbohydrate transition process. In addition, the neutron irradiation not only increased the total lipid production by 20% but also speeded up the reaction rate to reach the maximal total lipid production in 28 days. Our results can provide a better understanding of the lipid production in microalgae Chlorella sp. by neutron irradiation, which are critical for evaluation of neutron irradiation based technologies for the sustainable and renewable biofuel production.     

From piggery wastewater nutrients to biogas: Microalgae biomass revalorization through anaerobic digestion

Microalgae grown in swine wastewater were used as a promising strategy to produce renewable energy by coupling wastewater bioremediation and biomass revalorization. The efficiency of a microalgae consortium treating swine slurry at different temperature (15 and 23 °C) and illumination periods (11 and 14 h) was assessed for biomass growth and nutrient removal at two NH₄⁺ initial concentrations (80 and 250 mg L⁻¹ NH₄⁺). Favourable culture conditions (23 °C and 14 h of illumination) and high ammonium loads resulted in higher biomass production and greater nutrients removal rates. The initial N–NH₄⁺ load determined the removal mechanism, thus ammonia stripping and nitrogen uptake accounted similarly in the case of high NH₄⁺ load, while nitrogen uptake prevailed at low NH₄⁺ load. Under favourable conditions, nitrogen availability in the media determined the composition of the biomass. In this context, carbohydrate-rich biomass was obtained in batch mode while semi-continuous operation resulted in protein-rich biomass. The revalorization of the resultant biomass was evaluated for biogas production. Methane yields in the range of 106–146 and 171 ml CH₄ g COD⁻¹ were obtained for the biomasses grown in batch and semi-continuous mode, respectively. Biomass grown under favourable conditions resulted in higher methane yields and closer to the theoretically achievable.     

Effective extraction of microalgae lipids from wet biomass for biodiesel production

Producing biodiesel from lipid extracted from microalgae is a promising approach for sustainable fuel production. However, this approach is not yet commercialized due to the high costs of upstream processes that are associated with the time consuming and/or energy intensive drying, and lipid extraction processes. In this study, the possibility of avoiding the drying process, and extracting the lipid directly from the wet concentrated cells, using enzymatic disruption to enhance the extraction, has been tested. Results showed that lysozyme and cellulase were both efficient in disrupting cell walls and enhancing lipid extraction from wet samples, with highest lipid extraction yield of 16.6% achieved using lysozyme. The applicability of using supercritical CO₂ (SC-CO₂) in extracting lipid from wet biomass was also tested and the highest yield of 12.5% was achieved using lysozyme. In addition, a two-step culturing process was applied, using Scenedesmus sp., to combine both high biomass growth and lipid content. The strain was able to increase its biomass productivity in the first stage, reaching 174 mg l⁻¹ d⁻¹, with almost constant lipid content. In the second stage, the lipid content was enhanced by six-fold after three weeks of nitrogen starvation, but with lower biomass productivity.     

Assessment of triacylglycerol content in Chlorella vulgaris cultivated in a two-stage process

Chlorella vulgaris cultivation in two-stage process was applied to increase the lipid productivity without compromising the biomass productivity. At the first stage, microalgae was cultivated under nutrient sufficient conditions to obtain a maximized cell density; at the second stage, nitrate conditions are changed to trigger the accumulation of TAG. During first stage, the maximum biomass productivity (32 mg L⁻¹ d⁻¹) was observed after 13 days under nutrient sufficient conditions with 1.21 g L⁻¹ NaNO₃ and 0.00449 g L⁻¹ K₂HPO₄. Maximum lipid content (25.4% DW), lipid productivity (7.5 mg L⁻¹ d⁻¹) and TAG content (41.3% in total lipids) were favored by the nitrogen starvation conditions for more 4 days, at the second stage. Oil extracted at the second stage contained lower percentage of PUFAs being more suitable for the biodiesel production when compared with the oil extracted at the first stage. This two-stage phototrophic process is promising to provide a more efficient way for on a large-scale production of algal biomass and biodiesel production.     

Photosynthetic microbial desalination cell (PhMDC) using Chlamydomonas sp. (UKM6) and Scenedesmus sp. (UKM9) as biocatalysts for electricity production and water treatment

A current Microbial Desalination Cell (MDC) system often uses chemicals or air cathodes that are toxic and impractical for scalable applications. This work presents the MDC performance that utilized microalgae species Chlamydomonas sp. (UKM6) and Scenedesmus sp. (UKM9). These species supported the role of the terminal electron acceptor in the cathode chamber of photosynthetic MDCs (PhMDC). The results showed that PhMDC-UKM9 and UKM6 produced 1942 mW/m³ and 1714 mW/m³ power densities with 44% and 32% desalination rates, respectively. The desalination of salt concentration (35 g/L) was approaching the practical application of seawater. Both UKM6 and UKM9 achieved chemical oxygen demand (COD) removal in the anode chamber by 49% and 53%, respectively. 16S microbial community analysis in anolyte revealed phylum Firmicutes as the dominion community. This study demonstrated that the local microalgae species integrate power production, wastewater treatment, and water purification through PhMDC operation, comparable with other studies using commercial microalgae.     

Autotrophic cultivation of Botryococcus braunii for the production of hydrocarbons and exopolysaccharides in various media

Growth of Botryococcus braunii was studied using different autotrophic media such as bold basal medium (BBM), and bold basal with ammonium carbonate (BBMa), BG11, modified Chu 13 medium. Among the different autotrophic media used, BG11 was found to be the best medium for biomass and hydrocarbon production, although B. braunii showed appreciable level of growth and biomass production in all the tested media. The culture maintained at 16:8 h light and dark cycle with 1.2±0.2 klux light intensity at 25±1 °C temperature was found to be the best for growth (2.0 and 2.8 g L⁻¹ of biomass was produced by the B. braunii strains SAG 30.81 and LB-572, respectively) and hydrocarbon production (46% and 33%, respectively, by SAG 30.81 and LB 572 strains on dry weight basis) whereas continuous illumination with agitation at 90 rpm had maximum influence for the production of exopolysaccharides. The results of the present study indicate that the organism can acclimatize to different culture conditions and to a wide range of culture media with production of more than one metabolite.     

Carbon conversion and stabilisation of date palm and high rate algal pond (microalgae) biomass through slow pyrolysis

The processing of waste through pyrolysis technology is gaining momentum worldwide and is considered to be a green technology to reduce CO₂ emissions. This study is devoted to analysing the lignocellulosic biomass (date palm) and wastewater‐derived microalgae and the carbon‐rich char produced between temperature range (400°C‐600°C) from these biomass types. The properties of microalgae char showed that significant variation with date palm char exhibited high heating values (24‐28 MJ/kg), low ash content (11%‐16%), and high energy yield (48%‐42%). Algal biomass char showed considerably high nitrogen content (6%‐7%) as compared with date palm char (<1%), lower stability, and more significant influence on the price with respect to treatment temperature. Quaternary, pyrrolic, and pyridinic nitrogen species were found on the surface of the microalgae char, whereas no nitrogen species detected on date palm char due to low nitrogen content. The activation energy was also noted to be high for algal char during pyrolysis and combustion process. It can be concluded that date palm char is suitable for energy applications, whereas, algal char can be used for soil amendment, wastewater treatment, and applications requiring nitrogen‐doped char.