Cultivation of Chlorella sp. HS2 using wastewater from soy sauce factory.

Incorporation of wastewater from industrial sectors into the design of microalgal biorefineries has significant potential for advancing the practical application of this emerging industry. This study tested various food industrial wastewaters to assess their suitability for microalgal cultivation. Among these wastewaters, defective soy sauce (DSS) and soy sauce wastewater (SWW) were chosen but DSS exhibited the highest nutrient content with 13,500 ppm total nitrogen and 3051 ppm total phosphorus. After diluting DSS by a factor of 50, small-scale cultivation of microalgae was conducted to optimize culture conditions. SWW exhibited optimal growth at 25-30 °C and 300-500 µE m-2 s-1, while DSS showed optimal growth at 30-35 °C. Based on a 100-mL lab-scale and 3-L outdoor cultivation with an extended cultivation period, DSS outperformed SWW, exhibiting higher final biomass productivity. Additionally, nutrient-concentrated nature of DSS is advantageous for transportation at an industrial scale, leading us to select it as the most promising feedstock for microalgal cultivation. With further optimization, DSS has the potential to serve as an effective microalgal cultivation feedstock for large-scale biomass production.

Identification of Loliolide with Anti-Aging Properties from Scenedesmus deserticola JD052.

Herein, different extracts of Scenedesmus deserticola JD052, a green microalga, were evaluated in vitro as a potential anti-aging bioagent. Although post-treatment of microalgal culture with either UV irradiation or high light illumination did not lead to a substantial difference in the effectiveness of microalgal extracts as a potential anti-UV agent, the results indicated the presence of a highly potent compound in ethyl acetate extract with more than 20% increase in the cellular viability of normal human dermal fibroblasts (nHDFs) compared with the negative control amended with DMSO. The subsequent fractionation of the ethyl acetate extract led to two bioactive fractions with high anti-UV property; one of the fractions was further separated down to a single compound. While electrospray ionization mass spectrometry (ESI-MS) and nuclear magnetic resonance (NMR) spectroscopy analysis identified this single compound as loliolide, its identification has been rarely reported in microalgae previously, prompting thorough systematic investigations into this novel compound for the nascent microalgal industry.

Electric field and viscous fluid polarity effects on capillary-driven flow dynamics between parallel plates.

-Micropumps have attracted considerable interest in micro-electro-mechanical systems (MEMS), microfluidic devices, and biomedical engineering to transfer fluids through capillaries. However, improving the sluggish capillary-driven flow of highly viscous fluids is critical for commercializing MEMS devices, particularly in underfill applications. This study investigated the behavior of different viscous fluid flows under the influence of capillary and electric potential effects. We observed that upon increasing the electric potential to 500 V, the underfill flow length of viscous fluids increased by 45% compared to their capillary flow length. To explore the dynamics of underfill flow under the influence of an electric potential, the polarity of highly viscous fluids was altered by adding NaCl. The results indicated an increase of 20-41% in the underfill flow length of highly viscous conductive fluids (0.5-4% NaCl additives in glycerol) at 500 V compared to that at 0 V. The underfill viscous fluid flow length improved under the electric potential effect owing to the polarity across the substance and increased permittivity of the fluid. A time-dependent simulation, which included a quasi-electrostatic module, level set module, and laminar two-phase flow, was executed using the COMSOL Multiphysics software to analyze the effect of the external electric field on the capillary-driven flow. The numerical simulation results agreed well with the experimental data, with an average deviation of 4-7% at various time steps for different viscous fluids. Our findings demonstrate the potential of utilizing electric fields to control the capillary-driven flow of highly viscous fluids in underfill applications.

High-Resolution, Transparent, and Flexible Printing of Polydimethylsiloxane via Electrohydrodynamic Jet Printing for Conductive Electronic Device Applications.

In the field of soft electronics, high-resolution and transparent structures based on various flexible materials constructed via various printing techniques are gaining attention. With the support of electrical stress-induced conductive inks, the electrohydrodynamic (EHD) jet printing technique enables us to build high-resolution structures compared with conventional inkjet printing techniques. Here, EHD jet printing was used to fabricate a high-resolution, transparent, and flexible strain sensor using a polydimethylsiloxane (PDMS)/xylene elastomer, where repetitive and controllable high-resolution printed mesh structures were obtained. The parametric effects of voltage, flow rate, nozzle distance from the substrate, and speed were experimentally investigated to achieve a high-resolution (5 µm) printed mesh structure. Plasma treatment was performed to enhance the adhesion between the AgNWs and the elastomer structure. The plasma-treated functional structure exhibited stable and long strain-sensing cycles during stretching and bending. This simple printing technique resulted in high-resolution, transparent, flexible, and stable strain sensing. The gauge factor of the strain sensor was significantly increased, owing to the high resolution and sensitivity of the printed mesh structures, demonstrating that EHD technology can be applied to high-resolution microchannels, 3D printing, and electronic devices.

Flapping and powering characteristics of a flexible piezoelectric nanogenerator at Reynolds number range simulating ocean current.

For effective ocean energy harvesting, it is necessary to understand the coupled motion of the piezoelectric nanogenerator (PENG) and ocean currents. Herein, we experimentally investigate power performance of the PENG in the perspective of the fluid-structure interaction considering ocean conditions with the Reynolds number (Re) values ranging from 1 to 141,489. A piezoelectric polyvinylidene fluoride micromesh was constructed via electrohydrodynamic (EHD) jet printing technique to produce the ß-phase dominantly that is desirable for powering performance. Water channel was set to generate water flow to vibrate the flexible PENG. By plotting the Re values as a function of nondimensional bending rigidity (KB) and the structure-to-fluid mass ratio (M*), we could find neutral curves dividing the stable and flapping regimes. Analyzing the flow velocities between the vortex and surroundings via a particle image velocimetry, the larger displacement of the PENG in the chaotic flapping regime than that in the flapping regime was attributed to the sharp pressure gradient. By correlating M*, Re, KB, and the PENG performance, we conclude that there is critical KB that generate chaotic flapping motion for effective powering. We believe this study contributes to the establishment of a design methodology for the flexible PENG harvesting of ocean currents.

In vitro and in vivo assessment of squeeze-cast Mg-Zn-Ca-Mn alloys for biomedical applications.

Squeeze casting of biodegradable Mg-4Zn-0.5Ca-xMn (x = 0, 0.4 or 0.8 all in weight %) alloys is a process intended to improve mechanical properties (i.e., strength and ductility), corrosion performance (i.e., resistance), and biocompatibility (i.e., little to no cytotoxicity). In this study, we found that an increased Mn content causes the dendritic microstructure of squeeze-cast Mg-4Zn-0.5Ca-xMn to become more refined and uniform, while the volume fraction of the Ca2Mg6Zn3 phase simultaneously increases. Squeeze-cast Mg-4Zn-0.5Ca-0.8Mn presents the best yield strength, ultimate tensile strength, and elongation of the alloys tested. An electrochemical corrosion test in Hanks' solution at 36.5°C demonstrates that the corrosion resistance of squeeze-cast Mg-4Zn-0.5Ca-xMn alloys show improvement at higher Mn levels. Additionally, squeeze-cast Mg-4Zn-0.5Ca alloys containing Mn exhibit favorable biocompatibility, as evidenced by cell viability studies with MC3T3-E1 cells and a local lymph node assay test. Squeeze-cast alloy specimens implanted into the skull and spine of Sprague-Dawley rats for four weeks showed no serious cytotoxicity or foreign body response; however, swelling was observed in the implantation areas of Mn-free squeeze-cast Mg-4Zn-0.5Ca alloy, while no swelling was observed in rats implanted with Mn-containing Mg-4Zn-0.5Ca alloy. These findings indicate potential applications of biodegradable, Mn-containing, squeeze-cast Mg-4Zn-0.5Ca specimens in bone-reconstruction devices given their biocompatibility, mechanical properties, and degradation profiles. STATEMENT OF SIGNIFICANCE: Bioresorbable magnesium alloys have recently gained attention as viable biomaterials for skeletal reconstruction implants. Extensive research on biodegradable Mg alloy design, synthesis, and as-cast versus post-processed material properties useful for medical applications have been reported. The squeeze-casting technique used in this study can improve the mechanical properties (i.e., strengthening) and corrosive performance (reduced rate) of bioresorbable Mg-Zn-Ca-Mn alloys. Squeeze-casting of these alloys is also expected to improve specimen microstructure, near-net-shape manufacturing, and cost (i.e., reduced). This study provides an in vitro and in vivo assessment of squeeze-cast Mg-Zn-Ca-Mn alloys for biomedical applications.

Experimental, Theoretical, and Numerical Investigation of the Electric Field and Surface Wettability Effects on the Penetration Length in Capillary Flow.

This study addressed the dynamics of capillary-driven flow for different surface wettabilities by concentrating on the influence of electric potential. The capillary flow dynamics were investigated by varying the wettability (plasma-treated, hydrophobic, hydrophilic, and superhydrophilic) of a capillary surface, and the applied electric potential to the liquid ranged from 0 to 500 V. When an electric potential was applied to the liquid, the fluid flow penetration length increased by 30-50% due to the electrohydrodynamic (EHD)-driven flow by the Maxwell (electric) pressure gradient effect. The results showed that the EHD effect enhanced the fluid penetration through narrow gaps. The maximum fluid penetration was attained for every surface at 500 V, particularly for the superhydrophilic surface, which exhibited the highest value. The combined effect of the electric field and wettability resulted in an enhanced fluid penetration speed, reducing the underfill time. In addition, theoretical and numerical models were developed for comparison with the experimental results. The proposed models reinforce the observed fluid flow phenomenon on various surfaces under the influence of an electric field. These findings can provide alternative strategies for controlling the dynamic features of capillary imbibition by introducing an electric field and wettability effects, which has practical implications in flip-chip packaging, microfluidic devices, and the manipulation of biocells.

Application of Thromboelastographic Markers to Predict the Survival in Acute Trauma Patients.

BACKGROUND: The aim of the study is to investigate the coagulation status in trauma patients using thromboelastography and their association with survival and blood transfusion. METHODS: We included 452 trauma patients who visited the trauma center of Uijeongbu St. Mary's Hospital. The thromboelastography (TEG) clotting variables and routine coagulation tests were evaluated. Also, we investigated the transfusion requirement and mortality during hospitalization period. RESULTS: The mean age was 52.3 years and the mortality rate was 39/452 (8.6%). Lower GCS, longer TEG K-time, and lower TEG MA were independent factors associated with mortality. The lower MA group demonstrated the highest probability of survival (odds ratio 0.207), followed by prolonged R-time (odds ratio 0.220). The patient numbers in fibrinolysis shutdown (SD), physiologic fibrinolysis, and hyperfibrinolysis groups were 219 (52.3%), 131 (31.4%), and 68 (16.3%), respectively. The mortality rates of fibrinolysis SD group (11.9%) and hyperfibrinolysis (8.8%) were higher than the physiologic fibrinolysis groups (3.8%). The cutoff obtained from ROC analysis was found to be suitable for predicting survival. The transfusion requirements were significantly higher in the fibrinolysis SD group than in the other two groups. CONCLUSIONS: TEG based markers were shown to be more useful to make a diagnosis of coagulopathies including dysfibrinolysis and predict the survival than routine coagulation tests. Dysfunctional fibrinolysis showed higher mortality than physiologic group. If multiple integrations of each TEG markers are used, it would be helpful for prompt diagnosis and management of coagulopathies and to decrease preventable deaths in trauma.

Development of a novel nannochloropsis strain with enhanced violaxanthin yield for large-scale production.

BACKGROUND: Nannochloropsis is a marine microalga that has been extensively studied. The major carotenoid produced by this group of microalgae is violaxanthin, which exhibits anti-inflammatory, anti-photoaging, and antiproliferative activities. Therefore, it has a wide range of potential applications. However, large-scale production of this pigment has not been much studied, thereby limiting its industrial application. RESULTS: To develop a novel strain producing high amount of violaxanthin, various Nannochloropsis species were isolated from seawater samples and their violaxanthin production potential were compared. Of the strains tested, N. oceanica WS-1 exhibited the highest violaxanthin productivity; to further enhance the violaxanthin yield of WS-1, we performed gamma-ray-mediated random mutagenesis followed by colorimetric screening. As a result, Mutant M1 was selected because of its significant higher violaxanthin content and biomass productivity than WS-1 (5.21 ± 0.33 mg g- 1 and 0.2101 g L- 1 d- 1, respectively). Subsequently, we employed a 10 L-scale bioreactor to confirm the large-scale production potential of M1, and the results indicated a 43.54 % increase in violaxanthin production compared with WS-1. In addition, comparative transcriptomic analysis performed under normal light condition identified possible mechanisms associated with remediating photo-inhibitory damage and other key responses in M1, which seemed to at least partially explain enhanced violaxanthin content and delayed growth. CONCLUSIONS: Nannochloropsis oceanica mutant (M1) with enhanced violaxanthin content was developed and its physiological characteristics were investigated. In addition, enhanced production of violaxanthin was demonstrated in the large-scale cultivation. Key transcriptomic responses that are seemingly associated with different physiological responses of M1 were elucidated under normal light condition, the details of which would guide ongoing efforts to further maximize the industrial potential of violaxanthin producing strains.

High-Throughput In Vitro Screening of Changed Algal Community Structure Using the PhotoBiobox.

In a previous study, the sequential optimization and regulation of environmental parameters using the PhotoBiobox were demonstrated with high-throughput screening tests. In this study, we estimated changes in the biovolume-based composition of a polyculture built in vitro and composed of three algal strains: Chlorella sp., Scenedesmus sp., and Parachlorella sp. We performed this work using the PhotoBiobox under different temperatures (10-36°C) and light intensities (50-700 µmol/m-2/s-1) in air and in 5% CO2. In 5% CO2, Chlorella sp. exhibited better adaptation to high temperatures than in air conditions. Pearson's correlation analysis showed that the composition of Parachlorella sp. was highly related to temperature whereas Chlorella sp. and Scenedesmus sp. showed negative correlations in both air and 5% CO2. Furthermore, light intensity slightly affected the composition of Scenedesmus sp., whereas no significant effect was observed in other species. Based on these results, it is speculated that temperature is an important factor in influencing changes in algal polyculture community structure (PCS). These results further confirm that the PhotoBiobox is a convenient and available tool for performance of lab-scale experiments on PCS changes. The application of the PhotoBiobox in PCS studies will provide new insight into polyculture-based ecology.

Genome-wide high-throughput screening of interactive bacterial metabolite in the algal population using Escherichia coli K-12 Keio collection.

Algae-bacteria interaction is one of the main factors underlying the formation of harmful algal blooms (HABs). The aim of this study was to develop a genome-wide high-throughput screening method to identify HAB-influenced specific interactive bacterial metabolites using a comprehensive collection of gene-disrupted E. coli K-12 mutants (Keio collection). The screening revealed that a total of 80 gene knockout mutants in E. coli K-12 resulted in an approximately 1.5-fold increase in algal growth relative to that in wild-type E. coli. Five bacterial genes (lpxL, lpxM, kdsC, kdsD, gmhB) involved in the lipopolysaccharide (LPS) (or lipooligosaccharide, LOS) biosynthesis were identified from the screen. Relatively lower levels of LPS were detected in these bacteria compared to that in the wild-type. Moreover, the concentration-dependent decrease in microalgal growth after synthetic LPS supplementation indicated that LPS inhibits algal growth. LPS supplementation increased the 2,7-dichlorodihydrofluorescein diacetate fluorescence, as well as the levels of lipid peroxidation-mediated malondialdehyde formation, in a concentration-dependent manner, indicating that oxidative stress can result from LPS supplementation. Furthermore, supplementation with LPS also remarkably reduced the growth of diverse bloom-forming dinoflagellates and green algae. Our findings indicate that the Keio collection-based high-throughput in vitro screening is an effective approach for the identification of interactive bacterial metabolites and related genes.

Three new species of the genus Stephos Scott, 1892 (Crustacea, Copepoda, Calanoida, Stephidae) from Jeju Island, Korea.

During general field surveys carried out recently to collect benthopelagic copepods from near the substrate of the shallow waters off Jeju Island, Korea, a few specimens of three new species of Stephos Scott, 1892, were collected. The new species are placed in the genus Stephos because of the following combination of features: absence of seta on the basal exite of maxillule, and male right leg 5 ending in an unarmed claw-like and/or mitten-like segment. Stephos jejuensis sp. nov. can be distinguished from its congeners by body length 0.92 mm, left side of the female genital double-somite with protruding lobes, antennule that extends beyond the distal area of the genital double-somite, and the male leg 5 terminal complex. Stephos concavus sp. nov. can be distinguished from its congeners by the genital double-somite with protruding lobes on both sides, and the presence of larger spinules on the distomedial margin of leg 5. Stephos fortipes sp. nov. can be distinguished from its congeners by its longer body length, 1.12 mm long in the female, antennules that extend to the end of the genital double-somite, and the presence of a covered row of minute spinules on the ventral surface of the genital operculum in the female. Until now, 35 species of stephids were known worldwide.

Functional expression of polyethylene terephthalate-degrading enzyme (PETase) in green microalgae.

BACKGROUND: For decades, plastic has been a valuable global product due to its convenience and low price. For example, polyethylene terephthalate (PET) was one of the most popular materials for disposable bottles due to its beneficial properties, namely impact resistance, high clarity, and light weight. Increasing demand of plastic resulted in indiscriminate disposal by consumers, causing severe accumulation of plastic wastes. Because of this, scientists have made great efforts to find a way to biologically treat plastic wastes. As a result, a novel plastic degradation enzyme, PETase, which can hydrolyze PET, was discovered in Ideonella sakaiensis 201-F6 in 2016. RESULTS: A green algae, Chlamydomonas reinhardtii, which produces PETase, was developed for this study. Two representative strains (C. reinhardtii CC-124 and CC-503) were examined, and we found that CC-124 could express PETase well. To verify the catalytic activity of PETase produced by C. reinhardtii, cell lysate of the transformant and PET samples were co-incubated at 30 °C for up to 4 weeks. After incubation, terephthalic acid (TPA), i.e. the fully-degraded form of PET, was detected by high performance liquid chromatography analysis. Additionally, morphological changes, such as holes and dents on the surface of PET film, were observed using scanning electron microscopy. CONCLUSIONS: A PET hydrolyzing enzyme, PETase, was successfully expressed in C. reinhardtii, and its catalytic activity was demonstrated. To the best of our knowledge, this is the first case of PETase expression in green algae.

Biocompatible liquid-type carbon nanodots (C-paints) as light delivery materials for cell growth and astaxanthin induction of Haematococcus pluvialis.

In this study, we aimed to demonstrate the feasibility of the application of biocompatible liquid type fluorescent carbon nanodots (C-paints) to microalgae by improving microalgae productivity. C-paints were prepared by a simple process of ultrasound irradiation using polyethylene glycol (PEG) as a passivation agent. The resulting C-paints exhibited a carbonyl-rich surface with good uniformity of particle size, excellent water solubility, photo-stability, fluorescence efficiency, and good biocompatibility (<10.0 mg mL-1 of C-paints concentration). In the practical application of C-paints to microalgae culture, the most effective and optimized condition leading to growth promoting effect was observed at a C-paints concentration of 1.0 mg mL-1 (>20% higher than the control cell content). A C-paints concentration of 1-10.0 mg mL-1 induced an approximately >1.8 times higher astaxanthin content than the control cells. The high light delivery effect of non-cytotoxic C-paints was applied as a stress condition for H. pluvialis growth and was found to play a major role in enhancing productivity. Notably, the results from this study are an essential approach to improve astaxanthin production, which can be used in various applications because of its therapeutic effects such as cancer prevention, anti-inflammation, immune stimulation, and treatment of muscle-soreness.

Author Correction: Application of biosurfactant from Bacillus subtilis C9 for controlling cladoceran grazers in algal cultivation systems.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Two-stage cultivation strategy for the improvement of pigment productivity from high-density heterotrophic algal cultures.

Herein, a two-stage cultivation process was devised to overcome low pigment content of algal biomass grown in heterotrophy. Post-treatment conditions (i.e., light intensity, temperature, pH and salinity) were initially tested for dense heterotrophically-grown Chlorella sp. HS2 cultures in a multi-channel photobioreactor (mcPBR), and the results clearly indicated the influence of each abiotic factor on algal pigment production. Subsequently, the optimal post-treatment conditions (i.e., 455 µmol m-2 s-1, 34.8℃, pH 8.23 and 0.7% (w/v) salinity), in which highest accumulation of algal pigments is expected, were identified using Response Surface Methodology (RSM). Compared to the control cultures grown in mixotrophy for the same duration of entire two-stage process, the results indicated a significantly higher pigment productivity (i.e., 167.5 mg L-1 day-1) in a 5-L fermenter following the post-treatment at optimal conditions. Collectively, these results suggest that the post-treatment of heterotrophic cultures can be successfully deployed to harness the nascent algae-based bioeconomy.

Applicability of the masseter muscle as a nutritional biomarker.

Nutritional assessment is feasible with computed tomography anthropometry. The abdominal muscle at the L3 vertebra is a well-known nutritional biomarker for predicting the prognosis of various diseases, especially sarcopenia. However, studies on nutritional assessment of the brain using computed tomography are still scarce. This study aimed to investigate the applicability of the masseter muscle as a nutritional biomarker.Patients who underwent simultaneous brain and abdominopelvic computed tomography in the emergency department was retrospectively analyzed. We assessed their masseter muscle 2 cm below the zygomatic arch and abdominal muscle at L3 via computed tomography anthropometry. The skeletal muscle index, prognostic nutritional index, and other nutritional biomarkers were assessed for sarcopenia using the receiver operating characteristic curve analysis.A total of 314 patients (240 men and 72 women) were analyzed (mean age, 50.24 years; mean areas of the masseter and abdominal muscles, 1039.6 and 13478.3 mm, respectively). Masseter muscle areas significantly differed in sarcopenic, obese, and geriatric patients (P < .001). The areas under the curve of the masseter muscle in sarcopenic, geriatric, and obese patients were 0.663, 0.686, and 0.602, respectively. Multivariable linear regression analysis showed a correlation with the abdominal muscle area, weight, and age.The masseter muscle, analyzed via computed tomography anthropometry, showed a statistically significant association with systemic nutritional biomarkers, and its use as a nutritional biomarker would be feasible.

Redox-Active Tyrosine-Mediated Peptide Template for Large-Scale Single-Crystalline Two-Dimensional Silver Nanosheets.

Although self-assembly of various peptides has been widely applied, it is challenging to obtain single-crystalline and layer-by-layered nanostructures in a two-dimensional system. Here, we report a method for controlling the morphology and crystal growth at room temperature by a redox-active peptide template that can specifically co-assemble with metal ions. During the crystal growth, a silver ion-coordinated α-helical peptide (+3HN-YYACAYY-COO-) induces long-range atomic ordering at the air/water interface, which leads to multilayered single-crystalline silver nanosheets without an additional annealing process. Furthermore, this peptide template can facilitate efficient electron transfer between the independent metal nanosheets to improve electrochemical properties. We expect that this peptide template-based single-crystal growth method can be extended to synthesize other materials.

Monoterpenoid Loliolide Regulates Hair Follicle Inductivity of Human Dermal Papilla Cells by Activating the Akt/ß-Catenin Signaling Pathway.

Loliolide is one of the most ubiquitous monoterpenoid compounds found in algae, and its potential therapeutic effect on various dermatological conditions via agent-induced biological functions, including anti-oxidative and anti-apoptotic properties, was demonstrated. Here, we investigated the effects of loliolide on hair growth in dermal papilla (DP) cells, the main components regulating hair growth and loss conditions. For this purpose, we used a threedimensional (3D) DP spheroid model that mimics the in vivo hair follicle system. Biochemical assays showed that low doses of loliolide increased the viability and size of 3D DP spheroids in a dose-dependent manner. This result correlated with increases in expression levels of hair growth-related autocrine factors including VEGF, IGF-1, and KGF. Immunoblotting and luciferase-reporter assays further revealed that loliolide induced AKT phosphorylation, and this effect led to stabilization of ß-catenin, which plays a crucial role in the hair-inductive properties of DP cells. Further experiments showed that loliolide increased the expression levels of the DP signature genes, ALP, BMP2, VCAN, and HEY1. Furthermore, conditioned media from loliolide-treated DP spheroids significantly enhanced proliferation and the expression of hair growth regulatory genes in keratinocytes. These results suggested that loliolide could function in the hair growth inductivity of DP cells via the AKT/ ß-catenin signaling pathways.

Enhancement of ß-carotene production by regulating the autophagy-carotenoid biosynthesis seesaw in Chlamydomonas reinhardtii.

This work aimed to demonstrate a new strategy for enhancing the production of carotenoids through the regulation of seesaw cross-talk between autophagy and carotenoid biosynthesis pathways in Chlamydomonas reinhardtii. Autophagy-related ATG1 and ATG8 genes were first silenced using artificial microRNA, which in turn reduced the mRNA expression of ATG1 and ATG8 by 84.4% and 74.3%, respectively. While ATG1 kinase controls early step in autophagy induction and ATG8 is an essential factor for the downstream formation of autophagosome membranes, the decreased expression of these genes led to a 2.34-fold increase in the amount of ß-carotene content (i.e., 23.75 mg/g DCW). Furthermore, all mutants seemed to exhibit greater biodiesel properties than that of wild-type due to increased accumulation of saturated and monounsaturated fatty acids. These results support the role of autophagy in regulating the production of valuable metabolites, which could contribute to uplifting the economic outlook of nascent algal biorefinery.