The aim of this study was to construct Chlorella mutants deficient in chlorophyll synthesis by atmospheric pressure room temperature plasma (ARTP) mutagenesis, and screen novel algal species with very low chlorophyll content which is suitable for protein production by fermentation. Firstly, the lethal rate curve of mixotrophic wild type cells was established by optimizing the mutagenesis treatment time. The mixotrophic cells in early exponential phase were treated by the condition of over 95% lethal rate, and 4 mutants with the visual change of colony color were isolated. Subsequently, the mutants were cultured in shaking flasks heterotrophically for evaluation of their protein production performance. P. ks 4 mutant showed the best performance in Basal medium containing 30 g/L glucose and 5 g/L NaNO₃. The protein content and productivity reached 39.25% dry weight and 1.15 g/(L·d), with an amino acid score of 101.34. The chlorophyll a content decreased 98.78%, whereas chlorophyll b was not detected, and 0.62 mg/g of lutein content made the algal biomass appear golden yellow. This work provides a novel germplasm, the mutant P. ks 4 with high yield and high quality, for alternative protein production by microalgal fermentation.
Chlorella/metabolism* ; Chlorophyll A/metabolism* ; Plant Breeding ; Mutagenesis ; Chlorophyll/metabolism* ; Biomass ; Microalgae

Soluble cello-oligosaccharide with 2-6 oligosaccharide units is a kind of oligosaccharide with various biological functions, which can promote the proliferation of intestinal probiotics such as Bifidobacteria and Lactobacillus paracei. Therefore, it has a regulatory effect on human intestinal microbiota. In this study, a Cc 01 strain was constructed by expressing cellodextrin phosphorylase (CDP) in Escherichia coli. By combining with a previously constructed COS 01 strain, a three-enzyme cascade reaction system based on strains COS 01 and Cc 01 was developed, which can convert glucose and sucrose into cello-oligosaccharide. After optimization, the final titer of soluble cello-oligosaccharides with 2-6 oligosaccharide units reached 97 g/L, with a purity of about 97%. It contained cellobiose (16.8 wt%), cellotriose (49.8 wt%), cellotetrose (16.4 wt%), cellopentaose (11.5 wt%) and cellohexose (5.5 wt%). When using inulin, xylo-oligosaccharide and fructooligosaccharide as the control substrate, the biomass (OD₆₀₀) of Lactobacillus casei (WSH 004), Lactobacillus paracei (WSH 005) and Lactobacillus acidophilus (WSH 006) on cello-oligosaccharides was about 2 folds higher than that of the control. This study demonstrated the efficient synthesis of cello-oligosaccharides by a three-enzyme cascade reaction and demonstrated that the synthesized cello-oligosaccharides was capable of promoting intestinal microbial proliferation.
Humans ; Oligosaccharides ; Biomass ; Escherichia coli/genetics* ; Gastrointestinal Microbiome ; Glucose

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

This article summarized the reviews and research articles published in Chinese Journal of Biotechnology in the field of biomanufacturing in 2021. The article covered major chassis cells such as Escherichia coli, Bacillus subtilis, Corynebacterium glutamicum, Saccharomyces cerevisiae, filamentous fungi, non-model bacteria and non-conventional yeasts. Moreover, this article summarized the advances in the production of amino acids, organic acids, vitamins, higher alcohols, natural compounds (terpenoids, flavonoids, alkaloids), antibiotics, enzymes and enzyme-catalyzed products, biopolymers, as well as the utilization of biomass and one-carbon materials. The key technologies used in the construction of cell factories, such as regulation, evolution, and high-throughput screening, were also included. This article may help the readers better understand the R & D trend in biomanufacturing driven by engineered microbes.
Biomass ; Biotechnology ; Corynebacterium glutamicum/metabolism* ; Escherichia coli/metabolism* ; Metabolic Engineering ; Saccharomyces cerevisiae/genetics*

Tetrapyrrole compounds are a class of compounds with important functions. They exist in living organisms and have been widely used in agriculture, food, medicine, and other fields. The cumbersome process and high cost of chemical synthesis, as well as the shortcomings of unstable quality of animal and plant extraction methods, greatly hampered the industrial production and applications of tetrapyrrole compounds. In recent years, the rapid development of synthetic biology has provided new tools for microorganisms to efficiently synthesize tetrapyrrole compounds from renewable biomass resources. This article summarizes various strategies for the biosynthesis of tetrapyrrole compounds, discusses methods to improve its biosynthesis efficiency and future prospects, with the aim to facilitate the research on biosynthesis of tetrapyrrole compounds.
Biomass ; Plants/genetics* ; Synthetic Biology ; Tetrapyrroles

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

Lignin valorization for fuels and value-added products is essential to enhance the profitability and sustainability of biorefineries. Due to the complex and heterogeneous structure of lignin, technical barriers hinder the implementation of economic lignin utilization. Here, we summarize the major challenges facing lignin valorization processes. Different pretreatment methods, especially emerging combinatorial pretreatment approaches for isolating and tailoring lignin are introduced. To overcome the heterogeneity of lignin structure and improve lignin processability, advances in fractionation approaches including organosolv extraction, membrane technology, and gradient precipitation are analyzed and presented. Furthermore, progress in lignin valorization by thermochemical and biological conversion coupling with pretreatment and fractionation are systematically reviewed. Finally, we discuss advanced strategies and perspectives for future research involving biomass pretreatment, lignin fractionation and conversion processes.
Biomass ; Lignin

With the advantages of extensive sources, easy collection, renewability, high yield, carbon circulation, low pollution, and so on, Chinese medicinal solid waste can be converted into clean gas by pyrolysis and gasification, which is then able to serve for industrial production. This is of great practical significance in the context of energy shortage and for solid waste recycling in China. This paper reviews the research progress on biomass gasification principle, gasification medium, and reactor in gasification technology of Chinese medicinal solid waste in recent years. Meanwhile, based on the summary of related research, the defects and improvement measures regarding raw materials, gasification agents, by-products, and reactors were discussed, which provides direction for further development in the gasification technology of Chinese medicinal solid waste in the future.
Biomass ; China ; Pyrolysis ; Solid Waste ; Technology

BACKGROUND: Household air pollution from biomass fuels burning in traditional cookstoves currently appeared as one of the most serious threats to public health with a recent burden estimate of 2.6 million premature deaths every year worldwide, ranking highest among environmental risk factors and one of the major risk factors of any type globally. Improved cookstove interventions have been widely practiced as potential solutions. However, studies on the effect of improved cookstove interventions are limited and heterogeneous which suggested the need for further research. METHODS: A cluster randomized controlled trial study was conducted to assess the effect of biomass-fuelled improved cookstove intervention on the concentration of household air pollution compared with the continuation of an open burning traditional cookstove. A total of 36 clusters were randomly allocated to both arms at a 1:1 ratio, and improved cookstove intervention was delivered to all households allocated into the treatment arm. All households in the included clusters were biomass fuel users and relatively homogenous in terms of basic socio-demographic and cooking-related characteristics. Household air pollution was determined by measuring the concentration of indoor fine particulate, and the effect of the intervention was estimated using the Generalized Estimating Equation. RESULTS: A total of 2031 household was enrolled in the study across 36 randomly selected clusters in both arms, among which data were obtained from a total of 1977 households for at least one follow-up visit which establishes the intention-to-treat population dataset for analysis. The improved cookstove intervention significantly reduces the concentration of household air pollution by about 343 μg/m CONCLUSIONS: The biomass-fuelled improved cookstove intervention significantly reduces the concentration of household air pollution compared to the traditional method. This suggests that the implementation of these cookstove technologies may be necessary to achieve household air pollution exposure reductions. TRIAL REGISTRATION The trial project was retrospectively registered on August 2, 2018, at the clinical trials.gov registry database ( https://clinicaltrials.gov/ ) with the NCT03612362 registration identifier number.
Air Pollutants/analysis* ; Air Pollution, Indoor/prevention & control* ; Biomass ; Cluster Analysis ; Cooking/methods* ; Ethiopia

Production of biofuels such as ethanol from non-grain crops may contribute to alleviating the global energy crisis and reducing the potential threat to food security. Tobacco (Nicotiana tabacum) is a commercial crop with high biomass yield. Breeding of starch-rich tobacco plants may provide alternative raw materials for the production of fuel ethanol. We cloned the small subunit gene NtSSU of ADP-glucose pyrophosphorylase (NtAGPase), which controls starch biosynthesis in tobacco, and constructed a plant expression vector pCAMBIA1303-NtSSU. The NtSSU gene was overexpressed in tobacco upon Agrobacterium-mediated leaf disc transformation. Phenotypic analysis showed that overexpression of NtSSU gene promoted the accumulation of starch in tobacco leaves, and the content of starch in tobacco leaves increased from 17.5% to 41.7%. The growth rate and biomass yield of the transgenic tobacco with NtSSU gene were also significantly increased. The results revealed that overexpression of NtSSU gene could effectively redirect more photosynthesis carbon flux into starch biosynthesis pathway, which led to an increased biomass yield but did not generate negative effects on other agronomic traits. Therefore, NtSSU gene can be used as an excellent target gene in plant breeding to enrich starch accumulation in vegetative organs to develop new germplasm dedicated to fuel ethanol production.
Biomass ; Gene Expression Regulation, Plant ; Plant Breeding ; Plant Leaves/genetics* ; Plants, Genetically Modified/metabolism* ; Starch ; Tobacco/metabolism*