Dry Cell Weight Research Articles

Expanding the CRISPR Toolbox for Engineering Lycopene Biosynthesis in Corynebacterium glutamicum

Lycopene represents one of the central compounds in the carotenoid pathway and it exhibits a potent antioxidant ability with wide potential applications in medicine, food, and cosmetics. The microbial production of lycopene has received increasing concern in recent years. Corynebacterium glutamicum (C. glutamicum) is considered to be a safe and beneficial industrial production platform, naturally endowed with the ability to produce lycopene. However, the scarcity of efficient genetic tools and the challenge of identifying crucial metabolic genes impede further research on C. glutamicum for achieving high-yield lycopene production. To address these challenges, a novel genetic editing toolkit, CRISPR/MAD7 system, was established and developed. By optimizing the promoter, ORI and PAM sequences, the CRISPR/MAD7 system facilitated highly efficient gene deletion and exhibited a broad spectrum of PAM sites. Notably, 25 kb of DNA from the genome was successfully deleted. In addition, the CRISPR/MAD7 system was effectively utilized in the metabolic engineering of C. glutamicum, allowing for the simultaneous knockout of crtEb and crtR genes in one step to enhance the accumulation of lycopene by blocking the branching pathway. Through screening crucial genes such as crtE, crtB, crtI, idsA, idi, and cg0722, an optimal carotenogenic gene combination was obtained. Particularly, cg0722, a membrane protein gene, was found to play a vital role in lycopene production. Therefore, the CBIEbR strain was obtained by overexpressing cg0722, crtB, and crtI while strategically blocking the by-products of the lycopene pathway. As a result, the final engineered strain produced lycopene at 405.02 mg/L (9.52 mg/g dry cell weight, DCW) in fed-batch fermentation, representing the highest reported lycopene yield in C. glutamicum to date. In this study, a powerful and precise genetic tool was used to engineer C. glutamicum for lycopene production. Through the modifications between the host cell and the carotenogenic pathway, the lycopene yield was stepwise improved by 102-fold as compared to the starting strain. This study highlights the usefulness of the CRISPR/MAD7 toolbox, demonstrating its practical applications in the metabolic engineering of industrially robust C. glutamicum.

Optimizing microbioreactor cultivation strategies for Trichoderma reesei: from batch to fed-batch operations.

Filamentous fungi have long been recognized for their exceptional enzyme production capabilities. Among these, Trichoderma reesei has emerged as a key producer of various industrially relevant enzymes and is particularly known for the production of cellulases. Despite the availability of advanced gene editing techniques for T.reesei, the cultivation and characterization of resulting strain libraries remain challenging, necessitating well-defined and controlled conditions with higher throughput. Small-scale cultivation devices are popular for screening bacterial strain libraries. However, their current use for filamentous fungi is limited due to their complex morphology. This study addresses this research gap through the development of a batch cultivation protocol using a microbioreactor for cellulase-producing T.reesei strains (wild type, RutC30 and RutC30 TR3158) with offline cellulase activity analysis. Additionally, the feasibility of a microscale fed-batch cultivation workflow is explored, crucial for mimicking industrial cellulase production conditions. A batch cultivation protocol was developed and validated using the BioLector microbioreactor, a Round Well Plate, adapted medium and a shaking frequency of 1000rpm. A strong correlation between scattered light intensity and cell dry weight underscores the reliability of this method in reflecting fungal biomass formation, even in the context of complex fungal morphology. Building on the batch results, a fed-batch strategy was established for T.reesei RutC30. Starting with a glucose concentration of 2.5gl in the batch phase, we introduced a dual-purpose lactose feed to induce cellulase production and prevent carbon catabolite repression. Investigating lactose feeding rates from 0.3 to 0.75g(lh) , the lowest rate of 0.3g(lh) revealed a threefold increase in cellobiohydrolase and a fivefold increase in -glucosidase activity compared to batch processes using the same type and amount of carbon sources. We successfully established a robust microbioreactor batch cultivation protocol for T.reesei wild type, RutC30 and RutC30 TR3158, overcoming challenges associated with complex fungal morphologies. The study highlights the effectiveness of microbioreactor workflows in optimizing cellulase production with T.reesei, providing a valuable tool for simultaneous assessment of critical bioprocess parameters and facilitating efficient strain screening. The findings underscore the potential of microscale fed-batch strategies for enhancing enzyme production capabilities, revealing insights for future industrial applications in biotechnology.

Establishment of low-cost production platforms of polyhydroxyalkanoate bioplastics from Halomonas cupida J9.

Microbial production of polyhydroxyalkanoate (PHA) is greatly restricted by high production cost arising from high-temperature sterilization and expensive carbon sources. In this study, a low-cost PHA production platform was established from Halomonas cupida J9. First, a marker-less genome-editing system was developed in H. cupida J9. Subsequently, H. cupida J9 was engineered to efficiently utilize xylose for PHA biosynthesis by introducing a new xylose metabolism module and blocking xylonate production. The engineered strain J9UΔxylD-P8xylA has the highest PHA yield (2.81 g/L) obtained by Halomonas with xylose as the sole carbon source so far. This is the first report on the production of short- and medium-chain-length (SCL-co-MCL) PHA from xylose by Halomonas. Interestingly, J9UΔxylD-P8xylA was capable of efficiently utilizing glucose and xylose as co-carbon sources for PHA production. Furthermore, fed-batch fermentation of J9UΔxylD-P8xylA coupled to a glucose/xylose co-feeding strategy reached up to 12.57 g/L PHA in a 5-L bioreactor under open and unsterile condition. Utilization of corn straw hydrolysate as the carbon source by J9UΔxylD-P8xylA reached 7.0 g/L cell dry weight (CDW) and 2.45 g/L PHA in an open fermentation. In summary, unsterile production in combination with inexpensive feedstock highlights the potential of the engineered strain for the low-cost production of PHA from lignocellulose-rich agriculture waste.

Engineering Pseudomonas chlororaphis HT66 for the Biosynthesis of Copolymers Containing 3-Hydroxybutyrate and Medium-Chain-Length 3-Hydroxyalkanoates.

Polyhydroxyalkanoates (PHAs) are promising alternatives to petroleum-based plastics, owing to their biodegradability and superior material properties. Here, the controllable biosynthesis of scl-co-mcl PHA containing 3-hydroxybutyrate (3HB) and mcl 3-hydroxyalkanoates was achieved in Pseudomonas chlororaphis HT66. First, key genes involved in fatty acid β-oxidation, the de novo fatty acid biosynthesis pathway, and the phaC1-phaZ-phaC2 operon were deleted to develop a chassis strain. Subsequently, an acetoacetyl-CoA reductase gene phaB and a PHA synthase gene phaC with broad substrate specificity were heterologously expressed for producing and polymerizing the 3HB monomer with mcl 3-hydroxyalkanoates under the assistance of native β-ketothiolase gene phaA. Furthermore, the monomer composition of scl-co-mcl PHA was regulated by adjusting the amount of glucose and dodecanoic acid supplemented. Notably, the cell dry weight and scl-co-mcl PHA content reached 14.2 g/L and 60.1 wt %, respectively, when the engineered strain HT11Δ::phaCB was cultured in King's B medium containing 5 g/L glucose and 5 g/L dodecanoic acid. These results demonstrated that P. chlororaphis can be a platform for producing scl-co-mcl PHA and has the potential for industrial application.

Screening of efficient microalgae strains isolated from Tunisian ecosystems: Assessment of algal growth rate and added-value bioproducts for biorefinery applications

Microalgae are highly diverse organisms with significant applications in the production of high-value molecules and biodiesel from microalgae rich in oils. In this study, our main objectives were to evaluate the potential applications of four isolated microalgae strains, namely Scenedesmussp., Nannochloris sp.sp., Oscillatoria sp.sp., and Amphora sp.sp., which were selected from the natural environment. These strains were batch cultured, and their growth was monitored. We determined the content of chlorophyll a, carotenoids, and lipids, and analyzed the fatty acid profile of each cultivated strain to assess their lipid potential for biodiesel production. Our findings revealed that these strains exhibited high biomass production, ranging from 3000 mg. L−1 for Amphora sp.sp. to 3800 mg. L−1 for Oscillatoria sp.sp. Notably, Oscillatoria sp.sp. and Scenedesmus sp.sp. were found to be the richest in saturated fatty acids, with 10.68% and 9.36% of dry cell weight, respectively, suggesting their potential use in the production of biodiesel from microalgal oils with possibility of blending with other oils feedstock to improve biodiesel final quality. Furthermore, our results showed that the Nannochloris sp.sp. strain could be cultivated to extract pigments for various applications, as it exhibited high levels of chlorophyll a (38.43 mg. L−1) and carotenoids (4.153 μg. mL−1).

Integration of electronic nose, electronic tongue, and colorimeter in combination with chemometrics for monitoring the fermentation process of Tremella fuciformis

This study proposes an efficient method for monitoring the submerged fermentation process of Tremella fuciformis (T. fuciformis) by integrating electronic nose (e-nose), electronic tongue (e-tongue), and colorimeter sensors using a data fusion strategy. Chemometrics is employed to establish qualitative identification and quantitative prediction models. The Pearson correlation analysis was applied to extract features from the e-nose and tongue sensor arrays. The optimal sensor arrays for monitoring the deep fermentation process of T. fuciformis were obtained, and four different data fusion methods were developed by incorporating the colorimeter data features. To achieve qualitative identification, the physicochemical data and principal component analysis (PCA) results are utilized to determine three stages of the fermentation process. The fusion signal based on full features proves to be the optimal data fusion method, exhibiting the highest accuracy across different models. Notably, random forest (RF) is shown to be the most accurate pattern recognition method in this paper. For quantitative prediction, partial least squares regression (PLSR) and support vector regression (SVR) are employed to predict the sugar content and dry cell weight during fermentation. The best respective predictive R2 values for reducing sugar, tremella polysaccharide and dry cell weight are found to be 0.965, 0.988, and 0.970. Furthermore, due to its ability to capture nonlinear data relationships, SVR had superior performance in prediction modeling than PLSR. The results demonstrated that the combination of electronic sensor fusion signals and chemometrics provides a promising method for effectively monitoring T. fuciformis fermentation.

Molecular phylogenetics of the Umbelopsis genus-identification of new species and evaluation of their oil application value.

The aim of this study was to reconstruct the evolutionary framework of the genus Umbelopsis by using modern taxonomic strategies and evaluating the quality of oil and prospective uses of three distinct species. Three species of Umbelopsis were identified based on morphological characteristics and phylogenetic evidence obtained from three genes (ITS, LSU, and ACT). A new species of Umbelopsis was described and illustrated, and subsequently named U. ophiocordycipiticola. The characteristics of U. ophiocordycipiticola exhibited sporangia with a diameter ranging from 8 to 17µm. and sporangiospores that were oval to ellipsoidal in shape, irregularly angular, with dimensions of ∼1.9-2.9×1.7-3.0µm. Gas chromatography and mass spectrometry (GC-MS) were used to examine the composition of fatty acids. Notably, U. ophiocordycipiticola showed a significantly higher oil content of 50.89% in dry cell weight (DCW) compared to U. vinacea and U. ramanniana. The mean proportion of polyunsaturated fatty acids (PUFAs) in U. ophiocordycipiticola was 32.38%, and the maximum levels of γ-linolenic acid (GLA), arachidonic acid (ARA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) in U. ophiocordycipiticola were found to be 14.51, 0.24, 0.54, and 0.53%, respectively. The biodiesel quality from all three species complied with applicable standards set by the American Association for Testing and Materials (ASTM 6751) and the Brazilian National Petroleum Agency (ANP 255). The establishment of a novel species, U. ophiocordycipiticola, was strongly supported by morphological and molecular evidence. Umbelopsis ophiocordycipiticola exhibited a high-value PUFA content. Additionally, three Umbelopsis species demonstrated good quality for biodiesel production.

Polyhydroxybutyrate (PHB)-Based sustainable bioplastic derived from Bacillus sp. KE4 isolated from kitchen waste effluent

Petrochemical-derived plastics pose a major threat to the biosphere and the environment due to their acute or long-term toxicity. Therefore, to ensure sustainability, it is essential to find an effective replacement for synthetic plastics with natural and ecofriendly biomaterial. In this context, production of polyhydroxybutyrate (PHB) was produced by the Bacillus sp. KE4 using glucose as the carbon source, and a maximum dry cell weight (DCW) of 4.4 g/l with a PHB concentration of 3.1 g/l, PHB yield of 0.15 g/g was obtained, and the PHB content was 70.4% of the DCW. The extracted biopolymer was characterized using FTIR, GC-MS, 1H NMR, and 13C NMR which confirmed the structure of the biopolymer as PHB. The findings of TGA, DTG, DTA and DSC studies corresponded to PHB's excellent thermal stability. PHB biopolymer has a melting temperature of 168.7 °C and a maximum degradation temperature of 273 °C. The weight average molar mass (Mw), number average molar mass (Mn), and polydispersity index (PDI) of the extracted PHB was 1,55,436 g mol−1; 1,43,258 g mol−1; and 1.08, respectively. The XRD analysis validated the material's partly crystalline nature. FESEM-EDS and XPS were employed to determine the elemental composition of the purified PHB film. The Young's modulus, tensile strength and elongation at break showed rigid and brittle nature of PHB. Using open windrow composting method, the extracted PHB biopolymer film was assessed to determine its biodegradability. Within 28 days of soil burial, it was found that the biopolymer film had degraded by 61.34%. Cytotoxicity assessment in HepG2 cells indicated that PHB is non-cytotoxic in nature. The results have showed that the biomaterial is environmentally benign, biodegradable, and biocompatible; therefore, it may find its diverse application covering packaging, medicine, agriculture and allied fields.

Adsorption of Hg2+/Cr6+ by metal-binding proteins heterologously expressed in Escherichia coli

BackgroundRemoval of heavy metals from water and soil is a pressing challenge in environmental engineering, and biosorption by microorganisms is considered as one of the most cost-effective methods. In this study, the metal-binding proteins MerR and ChrB derived from Cupriavidus metallidurans were separately expressed in Escherichia coli BL21 to construct adsorption strains. To improve the adsorption performance, surface display and codon optimization were carried out.ResultsIn this study, we constructed 24 adsorption engineering strains for Hg2+ and Cr6+, utilizing different strategies. Among these engineering strains, the M’-002 and B-008 had the strongest heavy metal ion absorption ability. The M’-002 used the flexible linker and INPN to display the merRopt at the surface of the E. coli BL21, whose maximal adsorption capacity reached 658.40 μmol/g cell dry weight under concentrations of 300 μM Hg2+. And the B-008 overexpressed the chrB in the intracellular, its maximal capacity was 46.84 μmol/g cell dry weight under concentrations 500 μM Cr6+. While in the case of mixed ions solution (including Pb2+, Cd2+, Cr6+ and Hg2+), the total amount of ions adsorbed by M’-002 and B-008 showed an increase of up to 1.14- and 4.09-folds, compared to the capacities in the single ion solution.ConclusionThe construction and optimization of heavy metal adsorption strains were carried out in this work. A comparison of the adsorption behavior between single bacteria and mixed bacteria systems was investigated in both a single ion and a mixed ion environment. The Hg2+ absorption capacity is reached the highest reported to date with the engineered strain M’-002, which displayed the merRopt at the surface of chassis cell, indicating the strain’s potential for its application in practical environments.

Outdoor biohydrogen production by thermotolerant Rhodopseudomonas pentothenatexigens KKU-SN1/1 in a cluster of ten bioreactors system.

In tropical regions, the viability of outdoor photo-fermentative biohydrogen production faces challenges arising from elevated temperatures and varying light intensity. This research aimed to explore how high temperatures and outdoor environments impact both biohydrogen production and the growth of purple non-sulfur bacteria. Our findings revealed the potential of Rhodopseudomonas spp. as a robust outdoor hydrogen-producing bacteria, demonstrating its capacity to thrive and generate biohydrogen even at 40°C and under fluctuating outdoor conditions. Rhodopseudomonas harwoodiae NM3/1-2 produced the highest cumulative biohydrogen of 223mL/L under anaerobic light conditions at 40°C, while Rhodopseudomonas harwoodiae 2M had the highest dry cell weight of 2.93g/L. However, R. harwoodiae NM3/1-2 demonstrated the highest dry cell weight of 3.99g/L and Rhodopseudomonas pentothenatexigens KKU-SN1/1 exhibited the highest cumulative biohydrogen production of 400mL/L when grown outdoors. In addition, the outdoor enhancement of biohydrogen production was achieved through the utilization of a cluster of ten bioreactors system. The outcomes demonstrated a notable improvement in biohydrogen production efficiency, marked by the highest daily biohydrogen production of 493mL/Ld by R. pentothenatexigens KKU-SN1/1. Significantly, the highest biohydrogen production rate was noted to be 17 times greater than that observed in conventional batch production methods. This study is the first to utilize R. pentothenatexigens and R. harwoodiae for sustained biohydrogen production at high temperatures and in outdoor conditions over an extended operational period. The successful utilization of a clustered system of ten bioreactors demonstrates potential to scale-up for industrial biohydrogen production.

Streptococcus agalactiae serotype Ia capsular polysaccharide production from a Brazilian strain

The pathogen S. agalactiae, categorized as Lancefield group B streptococci (GBS), is a Gram-positive bacterium, encapsulated, anaerobic facultative bacillus, which occurs in pairs or short chains. It was responsible for more than 319,000 cases of infections in neonates. Early and late streptococci, in the course of an illness or medical condition, are infections that have high mortality and morbidity rates, with an incidence of 2.0 to 5.0 per 1000 live births in North America, Europe, and Australia. The infection of newborns usually happens through transmission from infected mothers during and after childbirth, with GBS being among the critical bacterial infections in the Brazilian epidemiological picture. S. agalactiae strains isolated from different regions of Brazil, indicate a great diversity among the isolated serotypes but with a higher incidence of serotype Ia. Publications report that, in pregnant women in Brazil, the prevalence of serotype Ia has been constant between 2002 and 2018, followed by serotypes II and IV. Serotypes Ib, III, and V also appear, but with some fluctuations in different regions of the country. The pathogen which causes invasive disease are usually surrounded by a polysaccharide capsule and is its major virulence factor, being that the key antigen in protective response triggered by polysaccharides and, more recently, protein-polysaccharide conjugate vaccines. The development of antibiotic resistance by these microorganisms to different antibiotics has been the guideline in leading therapeutic options against these infections including the use of prophylactic vaccines in pregnant women, immunosuppressed or older people. Such prophylactic alternatives can be constructed from capsular polysaccharides.Based on the experience in the production and purification of Neisseria meningitidis polysaccharides for vaccines, Bio-Manguinhos developed a process for the production, purification and characterization of S. agalactiae Ia polysaccharides. In the present study, the capsular polysaccharide from the strain isolated in clinical material of a public hospital, in Rio de Janeiro – Brazil, was obtained from simple batch cultures, with purification process achieving about 20 mg g−1 in yield for cell dry weight (CDW) and ca. of 100 % of purity. Hence, to prove that the CPS of GBS Ia can be consistently produced, purified, and characterized by NMR, HRGC-MS, and SEC-MALS – from such strain – to be used in vaccine productions against S. agalactiae.

Photosynthetic species composition determines bioplastics production in microbiomes: Strategy to enrich cyanobacteria PHB-producers

The aim of this study was to set the operating mode in regards to nutrients, temperature and light to use as a strategy to enrich a microbiome rich in cyanobacteria in polyhydroxybutyrate (PHB)-producers in order to enhance the biopolymer production. Alternate growth and accumulation phases were conducted for 197 days in a 3 L photobioreactor. Although, presence of green algae potentially reduced PHB production, the microbiome produced up to 22 % dry cell weight (dcw) PHB. Results highlighted the important role of culture composition and suggested that this methodology could be applied to a robust microbiome rich in cyanobacteria to boost PHB production.

Glycine betaine modulates extracellular polymeric substances to enhance microbial salinity tolerance

High salinity inhibits microbial activity in the bioremediation of saline wastewater. To alleviate osmotic stress, glycine betaine (GB), an osmoprotectant, is added to enhance the secretion of extracellular polymeric substances (EPS). These EPS are pivotal in withstanding environmental stressors, yet the intricate interplay between GB supplementation and microbial responses through EPS modifications—encompassing composition, molecular architecture, and electrochemical features—remains elusive in hypersaline conditions. Here we show microbial strategies for salinity endurance by investigating the impact of GB on the dynamic alterations of EPS properties. Our findings reveal that GB supplementation at 3.5% salinity elevates the total EPS (T-EPS) content from 12.50 ± 0.05 to 24.58 ± 0.96 mg per g dry cell weight. The observed shift in zeta potential from −28.95 to −6.25 mV at 0% and 3.5% salinity, respectively, with GB treatment, indicates a reduction in electrostatic repulsion and compaction. Notably, the EPS protein secondary structure transition from β-sheet to α-helix, with GB addition, signifies a more compact protein configuration, less susceptible to salinity fluctuations. Electrochemical analyses, including cyclic voltammetry (CV) and differential pulse voltammetry (DPV), reveal GB's role in promoting the release of exogenous electron shuttles, such as flavins and c-type cytochromes (c-Cyts). The enhancement in DPV peak areas (QDPV) with GB addition implies an increase in available extracellular electron transfer sites. This investigation advances our comprehension of microbial adaptation mechanisms to salinity through EPS modifications facilitated by GB in saline habitats.

Removal of sulfamethoxazole and production of cyanobacterial lipid promoted by the construction of a consortium containing a non-toxic cyanobacterium and sewage bacteria

This study evaluated the possibility of constructing a consortium containing sewage bacteria and cyanobacterial cells immobilized by Ca-alginate for the advanced treatment of antibiotic-polluted sewage coupled to cyanobacterial lipid production. Two non-toxic cyanobacterial species (Synechococcus sp. and Chroococcus sp.) were exposed to 0.5–20.0 μg/L of sulfamethoxazole (SMZ) for 4 days. Comparison between suspended and immobilized conditions indicated that immobilization increased (p < 0.05) SMZ removal percentage without reducing cyanobacterial growth performance (indicated by OD680), chlorophyll-a content, dry cell weight, lipid content and lipid weight/productivity of each cyanobacterium. Chroococcus sp. presented better antibiotic removal and lipid production ability than Synechococcus sp. After the addition of immobilized Chroococcus sp. into sewage, sewage bacteria attached to the surface and filled the pores of cyanobacterial beads. Immobilized Chroococcus sp. promoted the succession of sewage microbial community towards the dominance of Cellvibrio, Flavobacterium, Formivibrio and Pseudomonas, which participated in denitrification, phosphate accumulation and antibiotic removal. In the sewage microbial community, various functional genes upregulated by immobilized Chroococcus sp. directly showed negative correlation (p < 0.05) with sewage contaminants, including gltB, glnA and napA responsible for nitrogen removal; pstS responsible for phosphorus removal; nqrF, cbhA, glgC, malZ, pulA, uxaA and maeA responsible for COD removal; frmA, katG, maeA, uxaA, carB and cbhA responsible for SMZ removal. Coexisting sewage bacteria elevated (p < 0.05) lipid productivity of immobilized Chroococcus sp. to 29.13–36.52 mg/L/day through the upregulation of various genes related to metabolite exchange and signal transduction. Cyanobacterial cells degraded SMZ in collaboration with sewage bacteria, through deamination, nitration, bond cleavage, hydroxylation, desulfonation and nitrosation. The cyanobacteria-bacteria consortium presented 4-day removal percentages of 54.58–68.39%, 66.14–68.69%, 95.77–96.31% and 81.09–82.35% for SMZ, TN, TP and COD, respectively. In general, the cyanobacteria-bacteria consortium achieved a good combination of cyanobacterial lipid productivity and sewage treatment efficiency.

Production of iron enriched Saccharomyces boulardii: impact of process variables

About half of the 1.62 billion cases of anemia are because of poor diet and iron deficiency. Currently, the use of iron-enriched yeasts can be used as the most effective and possible way to prevent and treat anemia due to the ability of biotransformation of mineral compounds into the organic form. In this research, for the first time, Saccharomyces (S.) boulardii was used for iron enrichment with the aim that the probiotic properties of yeast provide a potential iron supplement besides improving the bioavailability of iron. Also, due to its higher resistance than other Saccharomyces strains against stresses, it can protect iron against processing temperatures and stomach acidic-enzymatic conditions. So, the effect of three important variables, including concentration of iron, molasses and KH2PO4 on the growth and biotransformation of yeast was investigated by the Box-Behnken design (BBD). The best conditions occurred in 3 g/l KH2PO4, 20 g/l molasses and 12 mg/l FeSO4 with the highest biotransformation 27 mg Fe/g dry cell weight (DCW) and 6 g/l biomass weight. Such yeast can improve fermented products, provide potential supplement, and restore the lost iron of bread, which is a useful iron source, even for vegetarians-vegans and play an important role in manage with anemia. It is recommended that in future researches, attention should be paid to increasing the iron enrichment of yeast through permeabilizing the membrane and overcoming the structural barrier of the cell wall.

Enhanced Microbial Protein Production from CO2 and Air by a MoS2 Catalyzed Bioelectrochemical System.

Carbon dioxide can be relatively easily reduced to organic matter in a bioelectrochemical system (BES). However, due to insufficient reduction force from in-situ hydrogen evolution, it is difficult for nitrogen reduction. In this study, MoS2 was firstly used as an electrocatalyst for the simultaneous reduction of CO2 and N2 to produce microbial protein (MP) in a BES. Cell dry weight (CDW) could reach 0.81±0.04 g/L after 14 d operation at -0.7 V (vs. RHE), which was 108±3 % higher than that from non-catalyst control group (0.39±0.01 g/L). The produced protein had a better amino acid profile in the BES than that in a direct hydrogen system (DHS), particularly for proline (Pro). Besides, MoS2 promoted the growth of bacterial cell on an electrode and improved the biofilm extracellular electron transfer (EET) by microscopic observation and electrochemical characterization of MoS2 biocathode. The composition of the microbial community and the relative abundance of functional enzymes revealed that MoS2 as an electrocatalyst was beneficial for enriching Xanthobacter and enhancing CO2 and N2 reduction by electrical energy. These results demonstrated that an efficient strategy to improve MP production of BES is to use MoS2 as an electrocatalyst to shift amino acid profile and microbial community.

Toxicological effects and mechanisms of lithium on growth, photosynthesis and antioxidant system in the freshwater microalga Chromochloris zofingiensis

The growing prevalence of lithium (Li) batteries has drawn public attention to Li as an emerging pollutant. The present study investigates the toxicity of Li+ on Chromochloris zofingiensis, examining physiological, biochemical and omics aspects. Results reveal hormesis effects of Li+ on C. zofingiensis growth. At Li+ concentrations below 5 mg L−1, Li+ can enhance chlorophyll content, mitochondrial activity, and antioxidant capacity, leading to increased dry cell weight and cell number. Conversely, when it exceeded 10 mg L−1, Li+ can reduce chlorophyll content, induce oxidative stress, and disrupt chloroplast and mitochondria structure and function, ultimately impeding cell growth. In addition, under 50 mg L−1 Li+ stress, microalgae optimize absorbed light energy use (increasing Fv/Fm and E TR ) and respond to stress by up-regulating genes in starch and lipid biosynthesis pathways, promoting the accumulation of storage components. Weighted gene co-expression network analysis indicates that peptidylprolyl cis/trans isomerase, GTPase and L-ascorbate oxidase might be the key regulators in response to Li+ stress. This research marks the toxic effects and molecular mechanisms of Li+ on freshwater microalga, which would improve our understanding of Li’s toxicology and contributing to the establishment of Li pollution standards.

Using the sulfide-oxidizing bacterium Geobacillus thermodenitrificans to restrict H2S release during chicken manure composting

Hydrogen sulfide (H2S) is a toxic gas massively released during chicken manure composting. Diminishing its release requires efficient and low cost methods. In recent years, heterotrophic bacteria capable of rapid H2S oxidation have been discovered but their applications in environmental improvement are rarely reported. Herein, we investigated H2S oxidation activity of a heterotrophic thermophilic bacterium Geobacillus thermodenitrificans DSM465, which contains a H2S oxidation pathway composed by sulfide:quinone oxidoreductase (SQR) and persulfide dioxygenase (PDO). This strain rapidly oxidized H2S to sulfane sulfur and thiosulfate. The oxidation rate reached 5.73 μmol min−1·g−1 of cell dry weight. We used G. thermodenitrificans DSM465 to restrict H2S release during chicken manure composting. The H2S emission during composting process reduced by 27.5% and sulfate content in the final compost increased by 34.4%. In addition, this strain prolonged the high temperature phase by 7 days. Thus, using G. thermodenitrificans DSM465 to control H2S release was an efficient and economic method. This study provided a new strategy for making waste composting environmental friendly and shed light on perspective applications of heterotrophic H2S oxidation bacteria in environmental improvements.

Effect of Iron Concentration on the Co-Production of Fucoxanthin and Fatty Acids in Conticribra weissflogii.

The production of fucoxanthin and fatty acids in Conticribra weissflogii has been examined, but there is still a lack of understanding regarding the impact of trace elements, including iron, on their co-production. To address this knowledge gap, this study investigated the effects of FeCl3·6H2O on the growth, fucoxanthin, and fatty acids of C. weissflogii. The findings revealed that the highest cell density (1.9 × 106 cells mL-1), cell dry weight (0.89 ± 0.15 g L-1), and total fatty acid concentration (83,318.13 µg g-1) were achieved at an iron concentration of 15.75 mg L-1, while the maximum carotenoid and fucoxanthin contents were obtained at an iron concentration of 3.15 mg L-1. The study demonstrated that the content of the active substance in C. weissflogii could be increased by adjusting the iron concentration, providing new information as to the more efficient co-production of fucoxanthin and fatty acids and offering experimental support for large-scale production.

Valorization of Cabbage Waste as a Feedstock for Microbial Polyhydroxyalkanoate Production: Optimizing Hydrolysis Conditions and Polyhydroxyalkanoate Production.

Establishing a platform for the bioconversion of waste resources into value-added compounds is critical for achieving a sustainable and eco-friendly economy. Herein, we produced polyhydroxyalkanoate via microbial fermentation using cabbage waste as a feedstock and metabolically engineered Escherichia coli. For this, the hydrolysis conditions of cabbage waste were optimized by focusing on parameters such as substrate and enzyme concentrations to enhance the saccharification efficiency. The phaABC operon, which encodes key enzymes responsible for polyhydroxyalkanoate biosynthesis in Ralstonia eutropha H16, was overexpressed in E. coli. Using cabbage hydrolysate as the feedstock, this engineered E. coli strain could produce poly(3-hydroxybutyrate) with a polymer content of 26.0 wt % of dry cell weight. Moreover, malic acid in cabbage hydrolysate significantly enhanced poly(3-hydroxybutyrate) production; the addition of 0.5 g/L malic acid markedly increased poly(3-hydroxybutyrate) content by 59.9%. This study demonstrates the potential of cabbage waste as a promising raw material for the microbial production of polyhydroxyalkanoate.