With the rapid development of synthetic biology, genetic engineering, and molecular manipulation methods in recent years, microalgae, as representatives of microbial cell factories, have been widely used as hosts in the production of high-value bioproducts, such as oils, pigments, proteins, and biofuels, demonstrating promising prospects of application in biochemical energy, food and drugs, and environmental protection. Despite these advancements, the low production efficiency of microalgae limits their industrial application. In addition to strain improvement and culture condition optimization, the regulation by exogenous chemical additives serves as a promising optimization strategy. This method relies on straightforward phenotypic screening and circumvents the necessity for intricate understanding of molecular targets in the metabolic and catabolic pathways involved in the synthesis of bioproducts. It enables rapid yield increasing of high-value bioproducts from microalgae and obtaining the required phenotypes. Although studies have reported the use of alternatives means such as exogenous additives to improve the growth of microalgae and the yield of high-value bioproducts, the classification and summarization of the types, applications, targeted strains, and molecular mechanisms of these additives are not comprehensive. Here, we review the studies using chemical inducers or enhancers to improve cell growth and high-value bioproduct accumulation in microalgae in recent years. This paper focuses on the types of exogenous additives, the effects of exogenous additives and their combinations on microalgae growth and high-value bioproduct accumulation, and the molecular mechanisms of related effects. We aim to provide information for researchers to use methods of synthetic biology to develop suitable cell chassis and harness microalgae for industrial production.
Microalgae/drug effects* ; Biofuels

To explore the differences in biological gas production in the waterlogged zone of a coal seam fire-affected area, in this study the in-situ gas production experiment was conducted with the mine water from aquitard layers in coal seam fire zones in Xinjiang. The results showed that the biogas production first increased and then decreased with the increase in distance, and the highest gas production reached 216.55 mL. The changes in key metabolic pathways during the anaerobic fermentation of coal were analyzed, which showed that as the distance from the aquitard layer in the coal seam fire zone increased, the methanogenesis pathways gradually shifted from acetic acid decarboxylation and carbon dioxide reduction to acetic acid decarboxylation and methylamine methanogenesis. The significant variability in the in-situ mine water reservoir conditions contributed to the differences. In addition, the reservoir pressure and temperature increased as the distance from the fire zone became longer, and the salinity of the farthest mine water in the reverse fault was the highest due to the lack of groundwater supply. Pearson correlation analysis revealed significant correlations of microbial communities with key functional genes and the types and concentrations of ions. The ions significantly influencing microbial enzymatic metabolic activities included Al³⁺, Fe²⁺, Co²⁺, Ni²⁺, Cu²⁺, Zn²⁺, Mg²⁺, PO₄^3-, and Mo⁶⁺. The differences in metabolic pathways were attributed to the integrated effects of a co-occurring environment with multiple ions. The gas production simulation experiments and metagenomic analyses provide data support for the practical application of in-situ biogas experiments, laying a foundation for engineering applications.
Biofuels ; Coal ; Methane/biosynthesis* ; Fires ; Groundwater ; Coal Mining ; Fermentation ; China ; Anaerobiosis

Cyanobacteria are the only prokaryotes capable of oxygenic photosynthesis, which have potential to serve as "autotrophic cell factories". However, the synthesis of biofuels and chemicals using cyanobacteria as chassis are suffered from poor stress tolerance and low yield, resulting in low economic feasibility for industrial production. Thus, it's urgent to construct new cyanobacterial chassis by means of synthetic biology. In recent years, adaptive laboratory evolution (ALE) has made great achievements in chassis engineering, including optimizing growth rate, increasing tolerance, enhancing substrate utilization and increasing product yield. ALE has also made some progress in improving the tolerance of cyanobacteria to high light intensity, heavy metal ions, high concentrations of salt and organic solvents. However, the engineering efficiency of ALE strategy in cyanobacteria is generally low, and the molecular mechanisms underpinning the tolerance to various stresses have not been fully elucidated. To this end, this review summarizes the ALE-associated technical strategies and their applications in cyanobacteria chassis engineering, following by discussing how to construct larger ALE mutation library, increase mutation frequency of strains and shorten evolution time. Moreover, exploration of the construction principles and strategies for constructing multi-stress tolerant cyanobacteria, and efficient analysis the mutant libraries of evolved strains as well as construction of strains with high yield and strong robustness are discussed, with the aim to facilitate the engineering of cyanobacteria chassis and the application of engineered cyanobacteria in the future.
Technology ; Photosynthesis/genetics* ; Cyanobacteria/genetics* ; Light ; Biofuels

Aims: This paper presents the report on biodiesel and biogas production at a laboratory scale from Scenedesmus strain. Methodology and results: Previously isolated and identified Scenedesmus were grown in 10 Liter flask using BG-11 media at 16 h light and 8 h dark cycle. Oven-dried biomass (20 g) from 16-day-old culture of Scenedesmus was finely grounded and subjected to lipids extraction by chloroform-methanol-NaCl mixture. Microalgal lipids (6 mL) were subjected to transesterification by using NaOH leading to the production of 5 mL biodiesel and 4 mL of glycerin. Biodiesel was rich in methyl esters of linoleic acid, phosphorothioc acid and dodecanoic acid, as shown by gas chromatography-mass spectrometry (GC-MS) analysis. Oven-dried microalgae (2 g) without lipid extraction and leftover biomass (2 g) after lipid extraction were subject to biogas production through anaerobic digestion. Biogas (34, 27 and 19 mL) were recorded respectively in oven-dried whole biomass; lipid extracted biomass and control over a period of 15 days of anaerobic digestion. Conclusion, significance and impact of study It was concluded that water bodies are rich in diverse algae, especially Scenedesmus sp., and this algae can be cultured to produce biodiesel and biogas. But the lipid accumulation potential of microalgae requires special treatment and lipid extraction methods are not up to the mark, which is a major bottleneck in biofuel production from microalgae.
Biofuels ; Scenedesmus--isolation & ; purification

Aims: The primary aim of this study was to utilize abundant palm oil mill effluent (POME) waste and turn it into a value-added product of biomass fuel with high calorific energy value (CEV) via fermentation and drying process, then simultaneously reduce abundant liquid waste. Methodology and results: POME is available abundantly in Malaysia and only a small portion of it is utilized to produce other value-added products. In this study, fermentation of POME in the presence of bacteria (Lysinibacillus sp.) and fungus (Aspergillus flavus) separately at 37 °C, 180 rpm for 5 days, followed by overnight oven-drying at 85 °C was conducted. Four fermentation medium conditions were performed, viz.: (1) autoclaved POME, (2) autoclaved POME with the addition of Lysinibacillus sp., (3) autoclaved POME with the addition of A. flavus and (4) POME as it is (non-sterile). Conclusion, significance and impact of study Among all conditions, fermentation utilizing autoclaved POME in the presence of A. flavus evinced the highest CEV of 25.18 MJ/kg. The fermentation in the presence of Lysinibacillus sp. strain revealed high COD and BOD removal efficiency of 59.20% and 320.44 mg/L as well as the highest reduction of oils and grease among other groups with the value of 15.84%. Future research directions are proposed for the elucidation of co-fermentation in the presence of both Lysinibacillus sp. and A. flavus.
Palm Oil ; Biomass ; Biofuels ; Waste Disposal, Fluid

Intense utilization and mining of fossil fuels for energy production have resulted in environmental pollution and climate change. Compared to fossil fuels, microalgae is considered as a promising candidate for biodiesel production due to its fast growth rate, high lipid content and no occupying arable land. However, monocultural microalgae bear high cost of harvesting, and are prone to contamination, making them incompetent compared with traditional renewable energy sources. Co-culture system induces self-flocculation, which may reduce the cost of microalgae harvesting and the possibility of contamination. In addition, the productivity of lipid and high-value by-products are higher in co-culture system. Therefore, co-culture system represents an economic, energy saving, and efficient technology. This review aims to highlight the advances in the co-culture system, including the mechanisms of interactions between microalgae and other microorganisms, the factors affecting the lipid production of co-culture, and the potential applications of co-culture system. Finally, the prospects and challenges to algal co-culture systems were also discussed.
Biofuels ; Biomass ; Coculture Techniques ; Flocculation ; Microalgae

Food wastes are rich in nutrients and can be used for producing useful chemicals through biotransformation. Some oleaginous microorganisms can use food wastes to produce lipids and high value-added metabolites such as polyunsaturated fatty acids, squalene, and carotenoids. This not only reduces the production cost, but also improves the economic value of the products, thus has large potential for commercial production. This review summarized the advances in food waste treatment, with a focus on the lipid production by oleaginous microorganisms using food wastes. Moreover, challenges and future directions were prospected with the aim to provide a useful reference for related researchers.
Biofuels ; Biotransformation ; Food ; Lipids ; Refuse Disposal

Biodiesel is an alternative fuel to addressing the energy shortage problem. Microbial lipids have attracted widespread attention as one of the potential feed-stocks for cost-effective and efficient biodiesel production. However, the large-scale production of microbial lipids is hampered by the complexity and the high cost of aseptic culturing approach. Metschnikowia pulcherrima is an oleaginous yeast with strong environmental adaptability. It is capable of utilizing a wide spectrum of substrates, and can be cultured under non-sterile conditions. Therefore, this yeast has great potential to replace the traditional oleaginous microorganisms, particularly in the area of recycling wastewater and solid waste for the production of biodiesel. Based on the analysis of lipid production and application conditions of M. pulcherrima, this review summarized the unique advantages of M. pulcherrima and the key factors affecting lipids production. We further discussed the feasibility of cultivating M. pulcherrima on various organic wastes under non-sterile conditions for lipids production. Moreover, we analyzed the challenges associated with M. pulcherrima's in the yield and mechanism for lipids production, and proposed perspectives for how to achieve efficient biodiesel production using this yeast.
Biofuels ; Candida ; Lipids ; Metschnikowia ; Yeasts

Lignocellulose can be hydrolyzed by cellulase into fermentable sugars to produce hydrogen, ethanol, butanol and other biofuels with added value. Pretreatment is a critical step in biomass conversion, but also generates inhibitors with negative impacts on subsequent enzymatic hydrolysis and fermentation. Hence, pretreatment and detoxification methods are the basis of efficient biomass conversion. Commonly used pretreatment methods of lignocellulose are chemical and physic-chemical processes. Here, we introduce different inhibitors and their inhibitory mechanisms, and summarize various detoxification methods. Moreover, we propose research directions for detoxification of inhibitors generated during lignocellulose pretreatment.
Biofuels ; Biomass ; Fermentation ; Hydrolysis ; Lignin/metabolism*

Yeast are comprised of diverse single-cell fungal species including budding yeast Saccharomyces cerevisiae and various nonconventional yeasts. Budding yeast is well known as an important industrial microorganism, which has been widely applied in various fields, such as biopharmaceutical and health industry, food, light industry and biofuels production. In the recent years, various yeast strains from different ecological environments have been isolated and characterized. Novel species have been continuously identified, and strains with diverse physiological characteristics such as stress resistance and production of bioactive compounds were selected, which proved abundant biodiversity of natural yeast resources. Genome mining of yeast strains, as well as multi-omics analyses (transcriptome, proteome and metabolome, etc.) can reveal diverse genetic diversity for strain engineering. The genetic resources including genes encoding various enzymes and regulatory proteins, promoters, and other elements, can be employed for development of robust strains. In addition to exploration of yeast natural diversity, phenotypes that are more suitable for industrial applications can be obtained by generation of a variety of genetic diversity through mutagenesis, laboratory adaptation, metabolic engineering, and synthetic biology design. The optimized genetic elements can be used to efficiently improve strain performance. Exploration of yeast biodiversity and genetic diversity can be employed to build efficient cell factories and produce biological enzymes, vaccines, various natural products as well as other valuable products. In this review, progress on yeast diversity is summarized, and the future prospects on efficient development and utilization of yeast biodiversity are proposed. The methods and schemes described in this review also provide a reference for exploration of diversity of other industrial microorganisms and development of efficient strains.
Biodiversity ; Biofuels ; Industrial Microbiology ; Metabolic Engineering ; Saccharomyces cerevisiae/genetics* ; Synthetic Biology