A comprehensive review on the heterotrophic production of bioactive compounds by microalgae.

Bioactive compounds derived from microalgae have garnered considerable attention as valuable resources for drugs, functional foods, and cosmetics. Among these compounds, photosynthetic pigments and polyunsaturated fatty acids (PUFAs) have gained increasing interest due to their numerous beneficial properties, including anti-oxidant, anti-viral, anti-bacterial, anti-fungal, anti-inflammatory, and anti-tumor effects. Several microalgae species have been identified as rich sources of bioactive compounds, including the Chlorophyceae Dunaliella and Haematococcus, the Bacillariophyta Phaeodactylum and Nitzschia, and the dinoflagellate Crypthecodinium cohnii. However, most of the reported microalgae species primarily grow through autotrophic mechanisms, resulting in low yields and high production costs of bioactive compounds. Consequently, the utilization of heterotrophic microalgae, such as Chromochloris zofingiensis and Nitzschia laevis, has shown significant advantages in the production of astaxanthin and eicosapentaenoic acid (EPA), respectively. These heterotrophic microalgae exhibit superior capabilities in synthesizing target compounds. This comprehensive review provides a thorough examination of the heterotrophic production of bioactive compounds by microalgae. It covers key aspects, including the metabolic pathways involved, the impact of cultivation conditions, and the practical applications of these compounds. The review discusses how heterotrophic cultivation strategies can be optimized to enhance bioactive compound yields, shedding light on the potential of microalgae as a valuable resource for high-value product development.

Ethanol extracts of Isochrysis zhanjiangensis alleviate acute alcoholic liver injury and modulate intestinal bacteria dysbiosis in mice.

BACKGROUND: Alcoholic liver disease (ALD) is responsible for 3.3 million deaths per annum. Efficacious therapeutic modalities or drug treatments for ALD have not yet been found, so it is urgent to seek new agents for preventing ALD and its related disease. Many experiments have indicated that modulating the gut microbiota and regulating the toll-like receptor 4 (TLR4)/nuclear transcription factor-κB (NF-κB) inflammatory pathway can provide a new target for prevention and treatment of ALD. Marine microalgae have their natural metabolic pathways to synthesize various of bioactive compounds as promising candidates for hepatoprotection. In this study, we investigated ethanol extracts from Isochrysis zhanjiangensis (EEIZ) to evaluate their ability to alleviate acute alcoholic liver injury, regulate TLR4/NF-κB inflammatory pathway and modulate intestinal bacteria dysbiosis in mice for ALD treatment. RESULTS: In the acute ALD mouse model, EEIZ reduced levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, triacylglyceride, total cholesterol and low-density lipoprotein, while increasing the level of high-density lipoprotein. Besides, TLR4, myeloid differentiation factor 88, NF-κB and tumor necrosis factor-α expression levels in liver tissue were effectively downregulated by EEIZ. Furthermore, treatment with EEIZ enhanced intestinal homeostasis and significantly alleviated the damage caused by alcohol. CONCLUSION: EEIZ showed effective hepatoprotective activity against alcohol-induced acute liver injury in mice as it could alleviate hepatocyte damage, suppress the TLR4/NF-κB inflammatory pathway and regulate the intestinal flora structure. EEIZ could be a good candidate for preventing acute alcoholic liver injury. © 2024 Society of Chemical Industry.

Preparation, multi-scale structures, and functionalities of acetylated starch: An updated review.

Acetylated starch has been widely used as food additives. However, there was limited information available regarding the impact of acetylation on starch structure and functionalities, as well as the advanced acetylation technologies. This review aimed to summarize current methods for starch acetylation and discuss the structure and functionalities of acetylated starch. Innovative techniques, such as milling, microwave, pulsed electric fields, ultrasonic, and extrusion, could be employed for environmental-friendly synthesis of acetylated starch. Acetylation led to the degradation of starch structures and weakening of the interactions between starch molecules, resulting in the disorganization of starch multi-scale ordered structure. The introduction of acetyl groups retarded the self-reassembly behavior of starch, leading to increased solubility, clarity, and softness of starch-based hydrogels. Moreover, the acetyl groups improved water/oil absorption capacity, emulsifiability, film-forming properties, and colonic fermentability of starch, while reduced the susceptibility of starch molecules to enzymes. Importantly, starch functionalities were largely influenced by the decoration of acetyl groups on starch molecules, while the impact of multi-scale ordered structures on starch physicochemical properties was relatively minor. These findings will aid in the design of structured acetylated starch with desirable functionalities.

Real-time response counterattack strategy of tolerant microalgae Chlorella vulgaris MBFJNU-1 in original swine wastewater and free ammonia.

This work was the first time to systematically clarify the potential tolerance mechanism of an indigenous Chlorella vulgaris MBFJNU-1 towards the free ammonia (FA) during the original swine wastewater (OSW) treatment by transcriptome analysis using C. vulgaris UETX395 as the control group. The obtained results showed that C. vulgaris MBFJNU-1 was found to be more resistant to the high levels of FA (115 mg/L) and OSW in comparison to C. vulgaris UETX395 (38 mg/L). Moreover, the transcriptomic results stated that some key pathways from arginine biosynthesis, electron generation and transmission, ATP synthesis in chloroplasts, and glutathione synthesis of C. vulgaris MBFJNU-1 were greatly related with the OSW and FA. Additionally, C. vulgaris MBFJNU-1 in OSW and FA performed similar results in the common differentially expressed genes from these mentioned pathways. Overall, these obtained results deliver essential details in microalgal biotechnology to treat swine wastewater and high free ammonia wastewater.

Microalgae-Derived Pigments for the Food Industry.

In the food industry, manufacturers and customers have paid more attention to natural pigments instead of the synthetic counterparts for their excellent coloring ability and healthy properties. Microalgae are proven as one of the major photosynthesizers of naturally derived commercial pigments, gaining higher value in the global food pigment market. Microalgae-derived pigments, especially chlorophylls, carotenoids and phycobiliproteins, have unique colors and molecular structures, respectively, and show different physiological activities and health effects in the human body. This review provides recent updates on characteristics, application fields, stability in production and extraction processes of chlorophylls, carotenoids and phycobiliproteins to standardize and analyze their commercial production from microalgae. Potential food commodities for the pigment as eco-friendly colorants, nutraceuticals, and antioxidants are summarized for the target products. Then, recent cultivation strategies, metabolic and genomic designs are presented for high pigment productivity. Technical bottlenecks of downstream processing are discussed for improved stability and bioaccessibility during production. The production strategies of microalgal pigments have been exploited to varying degrees, with some already being applied at scale while others remain at the laboratory level. Finally, some factors affecting their global market value and future prospects are proposed. The microalgae-derived pigments have great potential in the food industry due to their high nutritional value and competitive production cost.

Static and Dynamic Plantar Pressure Distribution in 94 Patients with Different Stages of Unilateral Knee Osteoarthritis Using the Footscan® Platform System: An Observational Study.

BACKGROUND Plantar pressure analysis is widely used in the study of knee osteoarthritis (KOA). The present study aimed to investigate the static and dynamic plantar pressure distribution in patients with different stages of unilateral KOA using the Footscan® platform system. MATERIAL AND METHODS We recruited 94 patients aged 61.75±7.23 years old with different stages of unilateral KOA for static and dynamic analysis using the Footscan® platform system. The static pressure (%) of the left, right, anterior, posterior, and the pelvic rotation (°) was assessed. The peak pressure (PP, kPa) was investigated in 10 areas of the foot: medial heel (MH), lateral heel (LH), midfoot (MF), first to fifth metatarsals (M1-M5), hallux (T1), and toes 2-5 (T2-5). The correlation between KOA stages and plantar pressure distributions was investigated. RESULTS The results revealed that static pressure on the unaffected side and pelvic rotation were positively correlated with KOA stages. In addition, there was a positive correlation between KOA stages and PP of M5, MF, and LH zones on the affected side and PP of M2, M3, and M4 zones on the unaffected side, and a negative correlation between KOA stages and PP of T1 and T2-5 zones on the affected side. CONCLUSIONS With the progression of KOA, static plantar pressure tends to distributed on the unaffected side, and the dynamic plantar pressure tends to be distributed laterally on both feet. The plantar pressure distributions in unilateral KOA patients are abnormal and are closely related to the severity of KOA.

Altering autotrophic carbon metabolism of Nitzschia closterium to mixotrophic mode for high-value product improvement.

An adaptive laboratory evolution (ALE) strategy was designed to evolve autotrophic Nitzschia closterium to mixotrophic growth for high productivity of essential amino acid (EAA), eicosapentaenoic acid (EPA) and fucoxanthin. The N. closterium growth was limited under glucose initially, but a red light emitting diode was innovatively applied to modify carbon metabolism and obtain mixotrophic strain of N. closterium GM. The N. closterium GM biomass concentration was improved by 65.07% comparing with wild type, but exhibited weak photosynthesis and strong glucose metabolism. At carbon metabolism levels, ALE promoted NADPH oxidase activity and induced protein degradation to lipid biosynthesis by elevating acetyl-CoA and pyruvate contents. It also improved carbon flux to TCA cycle, and elevated contents of glucose-6-phosphate, fructose-6-phosphate, glyceraldehyde-3-phosphate for providing sufficient ATP and NADPH. Productivities of EPA, EAA and fucoxanthin were increased by 41.0%, 18.8% and 20.4%, respectively. This ALE strategy was promising in microalgal production of high-value products.

Pilot-scale remediation of rare earth elements ammonium wastewater by Chlamydomonas sp. YC in summer under outdoor conditions.

This work evaluated the performance of real rare earth elements (REEs) wastewater purification and carbon dioxide (CO2) fixation by Chlamydomonas sp. YC with pilot-scale airlift-photobioreactors (AL-PBRs), tubular photobioreactors (TB-PBRs) and raceway ponds (ORWPs) under high-temperature outdoor conditions in summer. The obtained results showed that Chlamydomonas sp. YC at 1 g/L oyster shell piece (OSP) and 3 % CO2 had the highest biomass (1.9 g/L) and NH4+-N removal efficiency (34.0 %) during the REEs wastewater treatment. Among the selected photobioreactors, Chlamydomonas sp. YC to treat real REEs wastewater at 3 % CO2 under high-temperature outdoor conditions attained the highest biomass (2.3 g/L) in the TB-PBRs with the best NH4+-N removal efficiency (43.0 %). Furthermore, the input cost and CO2 net sequestration evaluation revealed that TB-PBRs was more economical photobioreactors to treat REEs wastewater and fix CO2 by Chlamydomonas sp. YC, providing some vital scientific details for REEs wastewater and CO2 fixation by microalgal biotechnology.

A novel very small granular starch from Chlorella sp. MBFJNU-17.

Novel resources of very small granular starch are of great interests to food scientists. We previously found Chlorella sp. MBFJNU-17 contained small granular starch but whether the MBFJNU-17 was a novel resource of very small granular starch remained unresolved. This study isolated and characterized the starch from MBFJNU-17 in comparison with quinoa starch (a typical very small granular starch), and discussed whether the MBFJNU-17 could be a resource of very small granular starch. Results showed that chlorella starch displayed a smaller size (1024 nm) than quinoa starch did (1107 nm), suggesting MBFJNU-17 was a good resource of very small granular starch. Additionally, chlorella starch had less amylose, higher proportion of long amylopectin branches, more ordered structures, thinner amorphous lamellae, better paste thermostability, and slower enzymatic digestion than quinoa starch did. These findings indicated that Chlorella sp. MBFJNU-17 was a novel resource of very small granular starch with desirable thermostability and nutritional attributes.

Fucoxanthin from marine microalgae: A promising bioactive compound for industrial production and food application.

Fucoxanthin attracts increasing attentions due to its potential health benefits, which has been exploited in several food commodities. However, fucoxanthin available for industrial application is mainly derived from macroalgae, and is not yet sufficiently cost-effective compared with microalgae. This review focuses on the strategies to improve fucoxanthin productivity and approaches to reduce downstream costs in microalgal production. Here we comprehensively and critically discuss ways and methods to increase the cell growth rate and fucoxanthin content of marine microalgae, including strain screening, condition optimization, design of culture mode, metabolic and genetic engineering, and scale-up production of fucoxanthin. The approaches in downstream processes provide promising alternatives for fucoxanthin production from marine microalgae. Besides, this review summarizes fucoxanthin improvements in solubility and bioavailability by delivery system of emulsion, nanoparticle, and hydrogel, and discusses fucoxanthin metabolism with gut microbes. Fucoxanthin production from marine microalgae possesses numerous advantages in environmental sustainability and final profits to meet incremental global market demands of fucoxanthin. Strategies of adaptive evolution, multi-stage cultivation, and bioreactor improvements have tremendous potentials to improve economic viability of the production. Moreover, fucoxanthin is promising as the microbiota-targeted ingredient, and nanoparticles can protect fucoxanthin from external environmental factors for improving the solubility and bioavailability.

Comparable effectiveness of transforaminal endoscopic spine system technique combined with selective nerve root block between far lateral lumbar disc herniation and central or paracentral herniation.

OBJECTIVES: This study aims to compare the clinical effectiveness of transforaminal endoscopic spine system (TESSYS) technique combined with selective nerve root block (SNRB) in treating patients with far lateral lumbar disc herniation (FLDH) and patients with central or paracentral herniation (C/PDH). PATIENTS AND METHODS: Between June 2015 and June 2019, a total of 204 patients (80 males, 124 females; mean age: 62.3±5.4 years; range, 51 to 66 years) with a herniated disc were included. Of these, 22 consecutive adult patients with FLDH formed the FLDH group, while 182 patients with C/PDH formed the C/PDH group. Considering that FLDH was a rare type of LDH and occurred outside the spinal canal, the patients with LDH in the spinal canal (C/PDH) were selected as the controls in our study. All cases received ultrasound-guided SNRB to identify the diseased disc and treated by the TESSYS technique. Data including demographics, duration of operation, duration of hospital stay, surgical cost, complications, Visual Analog Scale (VAS) scores for the back and leg, and Oswestry Disability Index (ODI) scores and the modified MacNab criteria were analyzed. RESULTS: The FLDH group presented the similar clinical outcomes and costs with the C/PDH group. No significant differences in the VAS score, ODI score, and Macnab score were observed between the groups (p>0.05 for all). Both groups showed the significantly improved postoperative VAS scores on Day 3, at 1, 3, 6, and 12 months compared to baseline. The postoperative ODI scores at 6 and 12 months were also significantly improved (p<0.05). At the final follow-up at 12 months, the FLDH group showed the MacNab criteria rating excellent and good of 81.8% and C/PDH group showed 84.62%. CONCLUSION: The FLDH patients presented the comparable clinical effectiveness with C/PDH patients. Based on these findings, the TESSYS technique combined with ultrasound-guided SNRB for FLDH is safe and feasible with caution, although the risk of nerve root injury may be worried.

The complete mitochondrial genome of Isochrysis galbana harbors a unique repeat structure and a specific trans-spliced cox1 gene.

The haptophyte Isochrysis galbana is considered as a promising source for food supplements due to its rich fucoxanthin and polyunsaturated fatty acids content. Here, the I. galbana mitochondrial genome (mitogenome) was sequenced using a combination of Illumina and PacBio sequencing platforms. This 39,258 bp circular mitogenome has a total of 46 genes, including 20 protein-coding genes, 24 tRNA genes and two rRNA genes. A large block of repeats (~12.7 kb) was segregated in one region of the mitogenome, accounting for almost one third of the total size. A trans-spliced gene cox1 was first identified in I. galbana mitogenome and was verified by RNA-seq and DNA-seq data. The massive expansion of tandem repeat size and cis- to trans-splicing shift could be explained by the high mitogenome rearrangement rates in haptophytes. Strict SNP calling based on deep transcriptome sequencing data suggested the lack of RNA editing in both organelles in this species, consistent with previous studies in other algal lineages. To gain insight into haptophyte mitogenome evolution, a comparative analysis of mitogenomes within haptophytes and among eight main algal lineages was performed. A core gene set of 15 energy and metabolism genes is present in haptophyte mitogenomes, consisting of 1 cob, 3 cox, 7 nad, 2 atp and 2 ribosomal genes. Gene content and order was poorly conserved in this lineage. Haptophyte mitogenomes have lost many functional genes found in many other eukaryotes including rps/rpl, sdh, tat, secY genes, which make it contain the smallest gene set among all algal taxa. All these implied the rapid-evolving and more recently evolved mitogenomes of haptophytes compared to other algal lineages. The phylogenetic tree constructed by cox1 genes of 204 algal mitogenomes yielded well-resolved internal relationships, providing new evidence for red-lineages that contained plastids of red algal secondary endosymbiotic origin. This newly assembled mitogenome will add to our knowledge of general trends in algal mitogenome evolution within haptophytes and among different algal taxa.

Transcriptome analysis of malate-induced Schizochytrium sp. FJU-512 reveals a novel pathway for biosynthesis of docosahexaenoic acid with enhanced expression of genes responsible for acetyl-CoA and NADPH accumulation.

Schizochytrium is one of the few oleaginous microalgae that produce docosahexaenoic acid (DHA)-rich lipids. In this study, global changes in gene expression levels of Schizochytrium sp. FJU-512 cultured with malate in a 15 l-bioreactor was analyzed using comparative transcriptomics. The changes were found mainly in the genes involved in oxidative phosphorylation, ß-oxidation, and pentose phosphate pathways. Consequently, the global changes in genes associated with the pathways could lead to an increase in the influx throughputs of pyruvate, branched-chain amino acids, fatty acids, and vitamin B6. Our transcriptome analysis indicated pyruvate dehydrogenase E2 component and acetolactate synthase I/II/III large subunit as major contributors to acetyl-CoA biosynthesis, whereas glucose-6-phosphate dehydrogenase was indicated as the major contributor to the biosynthesis of NADPH. An increase in DHA titer of up to 22% was achieved with the addition of malate to the fed-batch culture of Schizochytrium sp. FJU-512. This study provides an alternate method to enhance DHA production in Schizochytrium sp. FJU-512 through malate induced upregulation of genes responsible for acetyl-CoA and NADPH biosynthesis.

Dietary compounds slow starch enzymatic digestion: A review.

Dietary compounds significantly affected starch enzymatic digestion. However, effects of dietary compounds on starch digestion and their underlying mechanisms have been not systematically discussed yet. This review summarized the effects of dietary compounds including cell walls, proteins, lipids, non-starchy polysaccharides, and polyphenols on starch enzymatic digestion. Cell walls, proteins, and non-starchy polysaccharides restricted starch disruption during hydrothermal treatment and the retained ordered structures limited enzymatic binding. Moreover, they encapsulated starch granules and formed physical barriers for enzyme accessibility. Proteins, non-starchy polysaccharides along with lipids and polyphenols interacted with starch and formed ordered assemblies. Furthermore, non-starchy polysaccharides and polyphenols showed robust abilities to reduce activities of α-amylase and α-glucosidase. Accordingly, it can be concluded that dietary compounds lowered starch digestion mainly by three modes: (i) prevented ordered structures from disruption and formed ordered assemblies chaperoned with these dietary compounds; (ii) formed physical barriers and prevented enzymes from accessing/binding to starch; (iii) reduced enzymes activities. Dietary compounds showed great potentials in lowering starch enzymatic digestion, thereby modulating postprandial glucose response to food and preventing or treating type II diabetes disease.

Integrated green one-step strategy for concurrent recovery of phycobiliproteins and polyunsaturated fatty acids from wet Porphyridium biomass.

A novel and green one-step simultaneous extraction process of phycobiliproteins and polyunsaturated fatty acids (PUFAs) from wet Porphyridium biomass has been done and optimized by using three phase partitioning (TPP) process. Results showed that the coupling of ammonium sulfate and protein buoyancy-promoting t-butanol afforded the best TPP to extract phycobiliproteins and PUFAs in term of the extraction performance and cost-effectiveness. TPP process gave the best capability to simultaneously extract Porphyridium-derived phycobiliproteins and PUFAs in 20% ammonium sulfate, 0.5% biomass, and 1:0.5 slurry to t-butanol ratio at 100 rpm and 20 °C for 10 min of extraction time. Moreover, the established TPP system achieved excellent reproducibility in the extraction of Porphyridium biomass from different sources (Porphyridium cruentum and P. purpureum); and was successfully implemented in pilot-scale (20-L), indicating its industrial potential as a promising integrated approach to comprehensively exploit Porphyridium as a renewable bioresource for high-value bioproducts.

A newly isolated microalga Chlamydomonas sp. YC to efficiently remove ammonium nitrogen of rare earth elements wastewater.

The aim of this study was to establish a practical approach to remove ammonium nitrogen of rare earth elements (REEs) wastewater by an indigenous photoautotrophic microalga. Firstly, a new microalgal strain was successfully isolated from REEs wastewater and identified as Chlamydomonas sp. (named Chlamydomonas sp. YC). The obtained results showed that microalga could completely remove the NH4+-N of 10% REEs wastewater after 10 days of cultivation; however, the highest NH4+-N removal rate was attained by microalga to treat undiluted REEs wastewater. Then, three cultivation modes including batch, semi-continuous and continuous cultivation methods were developed to evaluate the ability of NH4+-N removal rate by this microalga to treat diluted (10%) and undiluted REEs wastewater. It was found that, Chlamydomonas sp. YC exhibited superior performance towards NH4+-N removal rates (32.75-61.05 mg/(L·d)) by semi-continuous and continuous processes for the treatments of 10% and undiluted REEs wastewater in comparison to the results (19.50-30.38 mg/(L·d) by batch process. Interestingly, under the same treatment conditions, among the three cultivation modes, microalga exhibited the highest removal rates of NH4+-N in undiluted REEs wastewater by semi-continuous (61.05 mg/(L·d)) and continuous (57.10 mg/(L·d) processes. In term of the biochemical analysis, microalgal biomass obtained from the wastewater treatment had 35.40-44.40% carbohydrate and 4.97-6.03% lipid, which could be potential ingredients for sustainable biofuels production. And the highest carbohydrate and lipid productivities attained by Chlamydomonas sp. YC in the continuous mode were 226.36 mg/(L·d) and 32.98 mg/(L·d), respectively. Taken together, the established processes mediated with Chlamydomonas sp. YC via continuous cultivation was the great promising approaches to efficiently remove NH4+-N of REEs wastewater and produce valuable biomass for sustainable and renewable biofuels in a simultaneous manner.

Long Non-Coding RNAs H19 and HOTAIR Implicated in Intervertebral Disc Degeneration.

Objective: Intervertebral disc degeneration (IDD) is the major cause of low back pain. We aimed to identify the key genes for IDD pathogenesis. Methods: An integrated analysis of microarray datasets of IDD archived in public Gene Expression Omnibus was performed. Bioinformatics analyses including identification of differentially expressed mRNAs/microRNAs/long non-coding RNAs (DEMs/DEMis/DELs), pathway enrichment, and competitive endogenous RNA (ceRNA) network construction were performed to give insights into the potential functions of differentially expressed genes (DEGs, including DEMs, DEMis, and DELs). The diagnostic value of DEMis in distinguishing IDD from normal controls was evaluated through receiver operating characteristic (ROC) analysis. Results: DEGs were identified in IDD, including H19 and HOTAIR. In the DEMis-DEMs network of IDD, miR-1291, miR-4270, and miR-320b had high connectivity with targeted DEMs. Cell death biological processes and the JAK-STAT pathway were significantly enriched from targeted DEMs. The area under the curve (AUC) of 10 DEMs including miR-1273e, miR-623, miR-518b, and miR-1291 in ROC analysis was more than 0.8, which indicated that those 10 DEMs had diagnostic value in distinguishing IDD from normal individuals. Conclusions: DELs H19 and HOTAIR were related to IDD pathogenesis. Cell death biological processes and the JAK-STAT pathway might play key roles in IDD development.

Chlorella vulgaris cultivation in pilot-scale to treat real swine wastewater and mitigate carbon dioxide for sustainable biodiesel production by direct enzymatic transesterification.

This study firstly addressed real swine wastewater (RSW) treatment by an indigenous Chlorella vulgaris MBFJNU-1 in 5-m3 outdoor open raceway ponds and then direct enzymatic transesterification of the resulting lipids from the wet biomass for sustainable biodiesel production. Compared to the control group, C. vulgaris MBFJNU-1 at 3% CO2 achieved higher microalgal biomass (478.5 mg/L) and total fatty acids content (21.3%), higher CO2 bio-fixation (63.2 mg/L/d) and lipid (9.1 mg/L/d) productivities, and greater nutrients removals (total nitrogen, 82.1%; total phosphorus, 28.4%; chemical oxygen demand, 37.1%). The highest biodiesel conversion (93.3%) was attained by enzymatic transesterification of wet disrupted Chlorella biomass with 5% lipase TL and 5% phospholipase PLA. Moreover, the enzymatic transesterification gave around 83% biodiesel conversion in a 15-L stirred tank bioreactor. Furthermore, the integrated process was a cost-effective approach to treat RSW and mitigate CO2 for microalgal biodiesel production, based on the mass and energy balances analysis.

A novel two-stage heterotrophic cultivation for starch-to-protein switch to efficiently enhance protein content of Chlorella sp. MBFJNU-17.

This work aimed to firstly establish an efficient and novel two-stage cultivation process to produce microalgal biomass rich in protein using a heterotrophic Chlorella sp. MBFJNU-17 strain. In the first-stage cultivation, to reduce the glucose and urea utilization, microalga achieved a high biomass at 40 g/L glucose and 1 g/L urea; meantime, the expression from starch biosynthesis genes of microalga was up-regulated under nitrogen-starvation conditions for starch accumulation (55.01%). In the second-stage cultivation, based on the over-compensation effect, Chlorella cells after the first-stage cultivation were further treated at 5 g/L glucose and 3 g/L urea to up-regulate starch degradation, central carbon metabolism and urea absorption genes expression to drive intracellular starch-to-protein switch for biosynthetic protein (59.75%). Moreover, microalga performed similar characteristics in a 10-L fermenter by the established process. Taken together, Chlorella sp. MBFJNU-17 was the promising candidate to produce high-value biomass enriched in protein by the established two-stage cultivation.

A novel and efficient strategy mediated with calcium carbonate-rich sources to remove ammonium sulfate from rare earth wastewater by heterotrophic Chlorella species.

This work was the first time to establish the desired approach with two heterotrophic Chlorella species for ammonium sulfate (AS)-rich rare earth elements (REEs) wastewater treatment by heterotrophic cultivation. The results showed that these two Chlorella species treated by 6 g/L CaCO3 performed the best ability to remove NH4+-N and SO42- of REEs wastewater. Moreover, the established process performed similar features in REEs wastewater treatment by replacing CaCO3 with eggshell powder (ESP) and oyster shell powder (OSP) enriched in CaCO3. Furthermore, microalgae treated by ESP/OSP in a 10-L fermenter showed 837.39 mg/(L·d) NH4+-N and 1,820 mg/(L·d) SO42- removal rates. The developed kinetic models could be well fitted to the experimental data obtained by the 10-L fermenter. Taken together, the established process mediated with two Chlorella species and ESP/OSP by heterotrophic cultivation was the great potential for AS-rich REEs wastewater treatment in a cost-effective manner.