1.Synthesis of cello-oligosaccharides which promotes the growth of intestinal probiotics by multi-enzyme cascade reaction.
Peng ZHENG ; Lei WANG ; Meirong HU ; Hua WEI ; Yong TAO
Chinese Journal of Biotechnology 2023;39(8):3406-3420
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
2.Breeding of Chlorella mutants deficient in chlorophyll synthesis and evaluation of its protein yield and quality.
Xiao CHEN ; Bailong WANG ; Dong WEI
Chinese Journal of Biotechnology 2023;39(3):1247-1259
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
3.Advances in the co-culture of microalgae with other microorganisms and applications.
Chang LI ; Wenxiang PING ; Jingping GE ; Yimeng LIN
Chinese Journal of Biotechnology 2022;38(2):518-530
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
4.Biomanufacturing driven by engineered microbes.
Chinese Journal of Biotechnology 2022;38(4):1267-1294
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*
5.Advances in the biosynthesis of tetrapyrrole compounds.
Fei PAN ; Yifan YAN ; Yifan ZHU ; Yi HU ; Xiaoqi XU ; Zheng XU ; Rui WANG ; Sha LI ; Hong XU ; Zhengshan LUO
Chinese Journal of Biotechnology 2022;38(4):1307-1321
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
6.Bio-valorization of palm oil mill effluent waste for the potential production of renewable biomass fuel pellets
Nurul Alia Syufina Abu Bakar ; Siti Baidurah
Malaysian Journal of Microbiology 2022;18(4):408-423
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
7.Biomass-fuelled improved cookstove intervention to prevent household air pollution in Northwest Ethiopia: a cluster randomized controlled trial.
Mesafint Molla ADANE ; Getu Degu ALENE ; Seid Tiku MERETA
Environmental Health and Preventive Medicine 2021;26(1):1-1
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
8.Progress in detoxification of inhibitors generated during lignocellulose pretreatment.
Li YANG ; Liping TAN ; Tongjun LIU
Chinese Journal of Biotechnology 2021;37(1):15-29
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*
9.Progress in the production of lignocellulolytic enzyme systems using Penicillium species.
Guodong LIU ; Liwei GAO ; Yinbo QU
Chinese Journal of Biotechnology 2021;37(3):1058-1069
The efficient production of lignocellulolytic enzyme systems is an important support for large-scale biorefinery of plant biomass. On-site production of lignocellulolytic enzymes could increase the economic benefits of the process by lowering the cost of enzyme usage. Penicillium species are commonly found lignocellulose-degrading fungi in nature, and have been used for industrial production of cellulase preparations due to their abilities to secrete complete and well-balanced lignocellulolytic enzyme systems. Here, we introduce the reported Penicillium species for cellulase production, summarize the characteristics of their enzymes, and describe the strategies of strain engineering for improving the production and performance of lignocellulolytic enzymes. We also review the progress in fermentation process optimization regarding the on-site production of lignocellulolytic enzymes using Penicillium species, and suggest prospect of future work from the perspective of building a "sugar platform" for the biorefinery of lignocellulosic biomass.
Biomass
;
Cellulase/metabolism*
;
Fermentation
;
Fungi/metabolism*
;
Lignin/metabolism*
;
Penicillium
10.Overexpression of NtAGPase small subunit gene increases leaf starch content and tobacco biomass.
Ying CHEN ; Yu GAO ; Teng LI ; Zhi XING ; Guiping CAI ; Jin'ai XUE ; Runzhi LI
Chinese Journal of Biotechnology 2021;37(8):2845-2855
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*


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