Dry Cell Weight Research Articles

Biofloc wastewater for microalgae (Chlorella ellipsoidea) production: an approach to algal biomass production and nutrient remediation

ABSTRACT Bold Basal Medium (BBM) is a standard but expensive medium for Chlorella ellipsoidea culture. This study evaluated the effect of biofloc wastewater (BWW) on the growth and pigment content of C. ellipsoidea and also investigated its bioremediation potential. The experiment was conducted for 24 d in five treatments, each with three replications. BBM was replaced with BWW at 0, 25, 50, 75, and 100% levels and considered as control (0% BWW), 25% BWW, 50% BWW, 75% BWW, and 100% BWW treatment, respectively. During the exponential period, the cell dry weight, optical density, cell density, specific growth rate, chlorophyll a, b, and β-carotene content were significantly (P ˂ 0.05) higher in 75% BWW treatment, which was followed by 0% BWW, 50% BWW, 100% BWW, and 25% BWW, correspondingly. NH3, NO2, and PO4 recovery efficiency were also higher in 75% BWW treatment. Therefore, the study proves that 75% BWW has the potential to culture C. ellipsoidea as an alternative to expensive BBM along with nutrient recovery from BWW.

Understanding exopolysaccharide byproduct formation in Komagataella phaffii fermentation processes for recombinant protein production

BackgroundKomagataella phaffii (Pichia pastoris) has emerged as a common and robust biotechnological platform organism, to produce recombinant proteins and other bioproducts of commercial interest. Key advantage of K. phaffii is the secretion of recombinant proteins, coupled with a low host protein secretion. This facilitates downstream processing, resulting in high purity of the target protein. However, a significant but often overlooked aspect is the presence of an unknown polysaccharide impurity in the supernatant. Surprisingly, this impurity has received limited attention in the literature, and its presence and quantification are rarely addressed.ResultsThis study aims to quantify this exopolysaccharide in high cell density recombinant protein production processes and identify its origin. In stirred tank fed-batch fermentations with a maximal cell dry weight of 155 g/L, the polysaccharide concentration in the supernatant can reach up to 8.7 g/L. This level is similar to the achievable target protein concentration. Importantly, the results demonstrate that exopolysaccharide production is independent of the substrate and the protein production process itself. Instead, it is directly correlated with biomass formation and proportional to cell dry weight. Cell lysis can confidently be ruled out as the source of this exopolysaccharide in the culture medium. Furthermore, the polysaccharide secretion can be linked to a mutation in the HOC1 gene, featured by all derivatives of strain NRRL Y-11430, leading to a characteristic thinner cell wall.ConclusionsThis research sheds light on a previously disregarded aspect of K. phaffii fermentations, emphasizing the importance of monitoring and addressing the exopolysaccharide impurity in biotechnological applications, independent of the recombinant protein produced.

High capacities of carbon capture and photosynthesis of a novel organic carbon-fixing microalgae in municipal wastewater: From mutagenesis, screening, ability evaluation to mechanism analysis

The development of wastewater treatment processes capable of reducing and fixing carbon is currently a hot topic in the wastewater treatment field. Microalgae possess a natural carbon-fixing advantage, and microalgae that can symbiotically coexist with indigenous bacteria in actual wastewater attract more significant attention. Ultraviolet (UV) mutagenesis and dissolved organic carbon (DOC) acclimation were applied to strengthen the carbon-fixing performance of microalgae in this study. The mechanisms associated with microalgal water purification ability, gene regulation at the molecular level and photosynthetic potential under different trophic modes resulting from carbon fixation and transformation were disclosed. The superior performance of Chlorella sp. MHQ2 was eventually screened out among a large number of mutants generated from 3 wild-type Chlorella strains. Results indicated that the dry cell weight of the optimal species Chlorella sp. HQ mutant MHQ2 was 1.91 times that of the wild strain in the pure algal system, more carbon from municipal wastewater (MW) were transferred to the microalgae and re-entered into the biological cycle through resource utilization. In addition, COD, NH3-N and TP removal efficiencies of MW by Chlorella sp. MHQ2 were found to increase to 95.8% (1.1-times), 96.4% (1.4-times), and 92.9% (1.2-times), respectively, under the extra DOC supply and the assistance of indigenous bacteria in the MW. In the transcriptome analysis of the logarithmic phase, the glycolytic pathway was inhibited, and the pentose phosphate pathway was mainly carried out for microalgal life activities, further promoting efficient energy utilization. Upon analysis of carbon capture capacity and photosynthetic potential in trophic mode, the addition of NaHCO3 increased the photosynthetic rate of Chlorella sp. MHQ2 in mixotrophy whereas it was attenuated in autotrophy. This study could provide a new perspective for the study of resource utilization and microalgae carbon- fixing mechanisms in the actual wastewater treatment process.

Enhancing astaxanthin accumulation through the expression of the plant-derived astaxanthin biosynthetic pathway in Dunaliella salina

Dunaliella salina, a microalga that thrives under high-saline conditions, is notable for its high β-carotene content and the absence of a polysaccharide cell wall. These unique characteristics render it a prime candidate as a cellular platform for astaxanthin production. In this study, our initial tests in an E. coli revealed that β-ring-4-dehydrogenase (CBFD) and 4-hydroxy-β-ring-4-dehydrogenase (HBFD) genes from Adonis aestivalis outperformed β-carotene hydroxylase (BCH) and β-carotene ketolase (BKT) from Haematococcus pluvialis counterparts by two-fold in terms of astaxanthin biosynthesis efficiency. Subsequently, we utilized electroporation to integrate either the BKT gene or the CBFD and HBFD genes into the genome of D. salina. In comparison to wild-type D. salina, strains transformed with BKT or CBFD and HBFD exhibited inhibited growth, underwent color changes to shades of red and yellow, and saw a nearly 50% decline in cell density. HPLC analysis confirmed astaxanthin synthesis in engineered D. salina strains, with CBFD + HBFD-D. salina yielding 134.88 ± 9.12 μg/g of dry cell weight (DCW), significantly higher than BKT-D. salina (83.58 ± 2.40 μg/g). This represents the largest amount of astaxanthin extracted from transgenic D. salina, as reported to date. These findings have significant implications, opening up new avenues for the development of specialized D. salina-based microcell factories for efficient astaxanthin production.

Formulation of biogenic fluorescent pigmented PHB nanoparticles from Rhodanobacter sp. for drug delivery.

Biogenic nanoparticles (NPs) have emerged as promising therapeutic formulations in effective drug delivery. Despite of various positive attributes, these NPs are often conjugated with various cytotoxic organic fluorophores for bioimaging, thereby reducing its effectiveness as a potential carrier. Herein, we aim to formulate biogenic fluorescent pigmented polyhydroxybutyrate (PHB) NPs from Rhodanobacter sp. strain KT31 (OK001852) for drug delivery. The bacterial strain produced 0.5g L-1 of polyhydroxyalkanoates (PHAs) from 2.04g L-1 of dry cell weight (DCW) under optimised conditions via submerged fermentation. Further, structural, thermal, and morphological charactersiation of the extracted PHAs was conducted using advance analytical technologies. IR spectra at 1719.25cm-1 confirmed presence of C = O functional group PHB. NMR and XRD analysis validated the chemical structure and crystallinity of PHB. TG-DTA revealed Tm (168 °C), Td (292 °C), and Xc (35%) of the PHB. FE-SEM imaging indicated rough surface of the PHB film and the biodegradability was confirmed from open windro composting. WST1 assay showed no significant cell death (> 50%) from 100 to 500µg/mL, endorsing non-cytotoxic nature of PHB. PHB NPs were uniform, smooth and spherical with size distribution and mean zeta potential 44.73nm and 0.5mV. IR and XRD peaks obtained at 1721.75cm-1 and 48.42Å denoted C = O and crystalline nature of PHB. Cell proliferation rate of PHB NPs was quite significant at 50µg/mL, establishing the non-cytotoxic nature of NPs. Further, in vitro efficacy of the PHB NPs needs to be evaluated prior to the biomedical applications.

Efficient β-carotene production in engineered Saccharomyces cerevisiae using simple sugars and agricultural waste-based carbon and nitrogen sources

β-carotene, a precursor to vitamin A, holds significant promise for health and nutrition applications. This study introduces an optimized approach for β-carotene production in Saccharomyces cerevisiae, leveraging metabolic engineering and a novel use of agricultural waste. The GAL80 gene deletion facilitated efficient β-carotene synthesis from sucrose, avoiding the costly galactose induction, and achieved titers up to 727.8 ± 68.0 mg/L with content levels of 71.8 ± 0.4 mg/g dry cell weight (DCW). Furthermore, the application of agricultural by-products, specifically molasses and fish meal as carbon and nitrogen sources, was investigated. This approach yielded a substantial β-carotene titer of 354.9 ± 8.2 mg/L and a content of 60.5 ± 4.3 mg/g DCW, showcasing the potential of these sustainable substrates for industrial-scale production. This study sets a new benchmark for cost-effective, green manufacturing of vital nutrients, demonstrating a scalable, eco-friendly alternative for β-carotene production.

Development of an H2 fuel cell electrochemical system powered by Escherichia coli cells

Because of the growing high importance of the development of biocatalytic fuel cell (FC) technologies for renewable energy-producing and testing systems for medical or environmental purposes, in this study, we constructed and demonstrated an H2 FC voltammeter working with graphite sample testing micro-strips and based on Escherichia coli microbial cells. Presented H2 FC voltammeter that provides fast and precise testing of bio-electrochemical possible reactions in biosamples for H2 and other gases, is automated with software which works in NI LabVIEW programming environment, has amplifier cascade system with high internal resistance, temperature controlling and resistance cascade. Microbial Hydrogenase (Hyd) enzymes reversibly catalyze the formation and oxidation of H2. Isolation and characterization of O2-tolerant [NiFe]-hydrogenases (Hyds) have given rise to new concepts in H2 FC. Escherichia coli and [NiFe]-Hyds can be applied as a biocatalyst anode in biofuel cells (BFCs). We evaluated the efficiency of applying the 3 µl (1.5 mg cell dry weight) E. coli intact cells or crude extracts on 0.5 cm2 as anode catalyzers in the bio-electrochemical system. The highest electrical potential (up to 0.7 V) was achieved with bacterial whole cells, which were grown on glucose and glycerol.

Production of polyhydroxybutyrate from industrial flue gas by microbial electrosynthesis

The aim of this work was to produce the biopolymer polyhydroxybutyrate (PHB) from industrial flue gas as CO2 source and electrolysis originating hydrogen via microbial electrosynthesis. Besides the laboratory experiments, the experiments were carried out under industrial conditions directly on-site in a cogeneration plant. We were able to demonstrate that the use of flue gas as a CO2 source has no detectable negative effect on bacterial growth and PHB production in comparison to a pure gas mixture. In an electrochemical H-cell 333 ± 44 mg L−1 PHB were obtained, which corresponds to a PHB content of 43 ± 3 % of the cell dry weight. By using flue gas for the production of the biopolymer PHB, not only CO2 emissions are reduced, but also possible environmental pollution from non-biodegradable plastics. To the best of our knowledge, this is the first time that the production of PHB from flue gas has been demonstrated using Cupriavidus necator.

Enhancing the biosynthesis of polyunsaturated fatty acids by Rhodotorula mucilaginosa and Lodderomyces elongisporus

Single-cell oils (SCO) produced by oleaginous yeast hold promise as a sustainable alternative for producing nutritionally and pharmaceutically valuable lipids. However, the accumulation of oils varies substantially between yeast spp. Consequently, identifying well-suited producers with a high innate capacity for lipids biosynthesis is paramount. Equally important is optimizing culturing and processing conditions to realize the total lipids production potential of selected strains. The marine Rhodotorula mucilaginosa and Lodderomyces elongisporus yeast were investigated to explore their potential for polyunsaturated fatty acids (PUFAs) production on high glucose media (HGM) using two-stage culture mode. Both strains accumulated > 20% (w/w) of their dry cell weight as lipids when grown on HGM using a two-stage culture system. Both yeast isolates exhibited a maximal lipid/biomass coefficient (YL/X) of 0.58–0.66 mg/mg at 7 °C and 0.49–0.53 mg/mg at 26 °C when grown on 8% glucose and produced monounsaturated and PUFAs similar to that of Menhaden and Salmon marine oils. For the first time, significant amounts of Eicosapentaenoic acid (19%) and Eicosadienoic acid (19.6%) were produced by L. elongisporus and R. mucilaginosa, respectively. Thus, the SCO derived from these wild strains possesses significant potential as a substitute source for the industrial-scale production of long-chain PUFAs, making them a promising contender in the market.Graphical

A self-stimulating system based on a polyhydroxyalkanoates coupled induction mechanism and its applications for Halomonas

Inducible gene expression systems are vital tools for synthetic biology. This study focused on a microbial polyhydroxyalkanoate (PHA) related inducible expression system, namely, the regulation mechanism of PHA synthesis associated protein PhaR and PhaP1 in Halomonas bluephagenesis which has employed to enhance PHA synthesis under open unsterile conditions. H. bluephagenesis strains ZS19 and ZS49 with one and two copies of the PHA synthesis system (PphaP1*-phaAB) were constructed based on the PHA self-stimulating synthesis mechanism. When grown in a 7-L bioreactor, H. bluephagenesis ZS19 and ZS49 were grown to 97.4 g/L and 89.6 g/L containing 76.3% and 77.1% PHA in the cell dry weights (CDW), with productivity of 2.06 g/L/h and 2.16 g/L/h, respectively. Compared with its control, their CDW, PHB percent and productivity increased by 20.4% and 10.8%, 13.5% and 14.7 %, and 36.4 % and 43.0 %, respectively. Furthermore, H. bluephagenesis ZS97, engineered with an elongated morphology induced by the PHA-mediated PhaR-phaP1 regulatory system, exhibited a 10 % increase in cell dry weight and a morphology approximately 10 times longer than the wild-type strain, allowing easy separation from the culture broth via centrifugation.

Unlocking growth potential in Halomonas bluephagenesis for enhanced PHA production with sulfate ions.

The mutant strain Halomonas bluephagenesis (TDH4A1B5P) was found to produce PHA under low-salt, non-sterile conditions, but the yield was low. To improve the yield, different nitrogen sources were tested. It was discovered that urea was the most effective nitrogen source for promoting growth during the stable stage, while ammonium sulfate was used during the logarithmic stage. The growth time of H. bluephagenesis (TDH4A1B5P) and its PHA content were significantly prolonged by the presence of sulfate ions. After 64 hr in a 5-L bioreactor supplemented with sulfate ions, the dry cell weight (DCW) of H. bluephagenesis weighed 132g/L and had a PHA content of 82%. To promote the growth and PHA accumulation of H. bluephagenesis (TDH4A1B5P), a feeding regimen supplemented with nitrogen sources and sulfate ions with ammonium sodium sulfate was established in this study. The DCW was 124g/L, and the PHA content accounted for 82.3% (w/w) of the DCW, resulting in a PHA yield of 101g/L in a 30-L bioreactor using the optimized culture strategy. In conclusion, stimulating H. bluephagenesis (TDH4A1B5P) to produce PHA is a feasible and suitable strategy for all H. bluephagenesis.

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.