Photosynthetic microalgae are emerging as potential biomass feedstock for sustainable production of biofuels and value-added bioproducts. CO2 biomitigation through these organisms is considered as an eco-friendly and promising alternative to the existing carbon sequestration methods. Nonetheless, the inherent relatively low photosynthetic capacity of microalgae has hampered the practical use of this strategy for CO2 biomitigation applications. Here, we demonstrate the feasibility of improving photosynthetic capacity by the genetic manipulation of the Calvin cycle in the typical green microalga Chlorella vulgaris. Firstly, we fused a plastid transit peptide to upstream of the enhanced green fluorescent protein (EGFP) and confirmed its expression in the chloroplast of C. vulgaris. Then we introduced the cyanobacterial fructose 1,6-bisphosphate aldolase, guided by the plastid transit peptide, into C. vulgaris chloroplast, leading to enhanced photosynthetic capacity (~ 1.2-fold) and cell growth. Molecular and physiochemical analyses suggested a possible role for aldolase overexpression in promoting the regeneration of ribulose 1,5-bisphosphate in the Calvin cycle and energy transfer in photosystems. Our work represents a proof-of-concept effort to enhance photosynthetic capacity by the engineering of the Calvin cycle in green microalgae. Our work also provides insights into targeted genetic engineering toward algal trait improvement for CO2 biomitigation uses.

Oleaginous microalgae have been considered promising sources of biodiesel due to their high lipid content. Nitrogen limitation/starvation is one of the most prominent strategies to induce lipid accumulation in microalgae. Nonetheless, despite numerous studies, the mechanism underlying this approach is not well understood. The aim of this study was to investigate the effect of nitrogen limitation and starvation on biochemical and morphological changes in the microalga Chlorella vulgaris FACHB-1068, thereby obtaining the optimal nitrogen stress strategy for maximizing the lipid productivity of microalgal biomass. The results showed that nitrogen limitation (nitrate concentration < 21.66 mg/L) and starvation enhanced the lipid content but generally decreased the biomass productivity, pigment concentration, and protein content in algal cells. Comparatively, 3-day nitrogen starvation was found to be a more suitable strategy to produce lipid-rich biomass. It resulted in an increased biomass production and satisfactory lipid content of 266 mg/L and 31.33%, respectively. Besides, nitrogen starvation caused significant changes in cell morphology, with an increase in numbers and total size of lipid droplets and starch granules. Under nitrogen starvation, saturated fatty acids (C-16:0, C-20:0, and C-18:0) accounted for the majority of the total fatty acids (~80%), making C. vulgaris FACHB-1068 a potential feedstock for biodiesel production. Our work may contribute to a better understanding of the biochemical and morphological changes in microalgae under nitrogen stress. Besides, our work may provide valuable information on increasing the lipid productivity of oleaginous microalgae by regulating nitrogen supply.

Miscanthus spp. are energy plants and excellent candidates for phytoremediation approaches of metal(loid)s-contaminated soils, especially when combined with plant growth-promoting bacteria. Forty-one bacterial strains were isolated from the rhizosphere soils and roots tissue of five dominant plants (Artemisia argyi Levl., Gladiolus gandavensis Vaniot Houtt, Boehmeria nivea L., Veronica didyma Tenore, and Miscanthus floridulus Lab.) colonizing a cadmium (Cd)-contaminated mining area (Huayuan, Hunan, China). We subsequently tested their plant growth-promoting (PGP) traits (e.g., production of indole-3-acetic acid, siderophore, and 1-aminocyclopropane-1-carboxylate deaminase) and Cd tolerance. Among bacteria, two strains, Klebsiella michiganensis TS8 and Lelliottia jeotgali MR2, presented higher Cd tolerance and showed the best results regarding in vitro growth-promoting traits. In the subsequent pot experiments using soil spiked with 10 mg Cd·kg⁻¹, we investigated the effects of TS8 and MR2 strains on soil Cd phytoremediation when combined with M. floridulus (Lab.). After sixty days of planting M. floridulus (Lab.), we found that TS8 increased plant height by 39.9%, dry weight of leaves by 99.1%, and the total Cd in the rhizosphere soil was reduced by 49.2%. Although MR2 had no significant effects on the efficiency of phytoremediation, it significantly enhanced the Cd translocation from the root to the aboveground tissues (translocation factor > 1). The combination of K. michiganensis TS8 and M. floridulus (Lab.) may be an effective method to remediate Cd-contaminated soils, while the inoculation of L. jeotgali MR2 may be used to enhance the phytoextraction potential of M. floridulus.

Bio-treatment of anaerobically treated swine wastewater (ADSW) mediated by microalgae has been deemed as a promising strategy. In the present study, six microalgal strains were used to conduct batch experiments in 0~100% ADSW in order to evaluate their potentials for nutrient removal and biodiesel production. Two strains, Chlorella vulgaris FACHB-8 and Chlorella sp. FACHB-31, were selected based on their better growth performances, higher tolerance to wastewater (up to 100%), and better nutrient removal abilities. The capacity of each strain to remove TN, TP, NH₄⁺-N, as well as lipid production and biomass composition in 100% ADSW were further examined. After 15 days of culture, 87.68~89.85%, 92.61~93.68%, and 97.02~97.86% of the nitrogen, phosphorus, and ammonia nitrogen were removed by Chlorella sp. FACHB-31 and C. vulgaris FACHB-8. Their lipid content and lipid productivities were 29.63~33.33% and 18.91~23.10 mg L⁻¹ d⁻¹, respectively. Proteins were both the major biomass fraction followed by lipids and then carbohydrates. Their fatty acid profiles both mainly consisted of C-16:0, C-18:1, C-18:0, and C-18:2. Taken together, our results suggest that C. vulgaris FACHB-8 and Chlorella sp. FACHB-31 are potential candidates for biodiesel production by using ADSW as a good feedstock.

This study investigated anaerobically digested swine wastewater (ADSW) as a nutrient source for Chlorella vulgaris FACHB-8 cultivation under mixotrophic conditions with carbon supplementation. The microalgal strain was grown in ADSW supplemented with six carbon sources, followed by concentration optimization. Under optimized conditions (20 g/L glucose), FACHB-8 demonstrated a high biomass productivity (271.31 mg/L/day) and a specific growth rate of 0.42 per day. The system achieved an 88.70% total nitrogen removal and an 82.93% total phosphorus removal. The biomass contained 45.59% lipids, 29.72% proteins, and 13.05% carbohydrates, with fatty acid methyl esters showing balanced proportions of saturated (50.77%) and unsaturated fatty acids (49.23%). These findings highlight the potential of glucose-based mixotrophic cultivation for simultaneous wastewater treatment, renewable biomass production, and value-added lipid production. This work proposes a scalable swine wastewater treatment system that synergizes bioremediation and renewable energy production via carbon-enhanced microalgae cultivation, offering a dual-functional strategy for sustainable livestock wastewater reuse.