Biotechnologia最新文献

The exploitation of food industry wastes and their conversion into value-added products present a promising and continuously growing field, given the diversity of elaborated wastes. The current work aimed to utilize sweet cheese whey as a growth medium for acid protease production by a local fungus strain. The biochemical and physicochemical properties of the cheese whey, such as pH, conductivity, chemical oxygen demand, biological oxygen demand (BOD₅), total nitrogen and protein contents, and mineral salts, were assessed using various analytical methods. The effect of certain parameters on acid protease production by Mucor circinelloides MG603064.1 through the SmF process was investigated using the conventional design method "One factor at a time". Subsequent to characterization, the crude extract was used in a trial to create fromage frais, compared to the commercial rennin CHY-MAX^® Powder Extra. Cheese whey characterization revealed its richness in total nitrogen (1.044 ± 0.044 g/l), protein content (6.52 ± 0.04 g/l), and principal mineral salts: calcium (1.637 ± 0.037 g/l), phosphorus (1.173 ± 0.023 g/l), and chloride (1.66 ± 0.09 g/l). The optimal values of the SmF process for acid protease production, such as the inoculum size, beef extract, and KH₂PO₄ supplements, the initial pH of cheese whey, and incubation temperature were, respectively, 11% (v/v), 0.4% (w/v), 0.5% (w/v), 5.5, and 30°C. Under these conditions, the lowest milk-clotting time of 290 s was achieved, representing an 18.41-fold increase compared to the initial step using the unoptimized medium. The enzyme showed maximum milk-clotting activity at pH 5, a temperature of 60°C, and in the presence of 0.025 M of CaCl₂. The enzyme activity also significantly improved with sonication (35 kHz) for 10 min. The crude extract of M. circinelloides ensured the production of fresh cheese samples with characteristics roughly similar to those obtained by the control (CHY-MAX^® rennin). The acid protease of M. circinelloides could successfully substitute the conventional rennin in the manufacture of fresh cheese.

The seed germination dynamics of Acacia nilotica, Bauhinia rufescens, Faidherbia albida, and Piliostigma reticulatum were investigated over 28 days. Seeds were pretreated with concentrated sulfuric acid. Determined germination parameters included germination energy, germination period, germination capacity, germination inertia, and viability loss. Seeds exposed to sulfuric acid for extended periods (30, 40, 50, and 60 min) exhibited a higher germination rate (α = 0.05). For A. nilotica seeds, the 50 min acid treatment resulted in the highest germination energy of 85.5% and germination capacity of 89.5% (P = 0.001); conversely, the 60-min treatment yielded the highest germination energy and capacity, both 96.5% (P = 0.079), for P. reticulatum. In the case of B. rufescens, the 30-min treatment led to the highest germination energy of 93% and germination capacity of 88% (P = 0.001). For F. albida, all acid treatments resulted in 100% for both germination energy and germination capacity (P = 0.621). Viability losses for A. nilotica and P. reticulatum were higher (32 and 30%, respectively) than those for B. rufescens and F. albida, which were 19.5 and 6%, respectively (P = 0.000). Generally, higher germination energy resulted in lower viability loss, dependent on the species. Analyses of germination inertia and viability loss suggest that seeds of A. nilotica and P. reticulatum possess a greater ability to survive in arid land climates than those of B. rufescens and F. albida. However, due to the advantage of lower viability loss, B. rufescens and F. albida should be preferred for the natural restoration of arid land ecosystems where seed availability is a major concern.

New prophylactic vaccine platforms are imperative to combat respiratory infections. The efficacy of T and B memory cell-mediated protection, generated through the adenoviral vector, was tested to assess the effectiveness of the new adenoviral-based platforms for infectious diseases. A combination of adenovirus AdV1 (adjuvant), armed with costimulatory ligands (ICOSL and CD40L), and rRBD (antigen: recombinant nonglycosylated spike protein rRBD) was used to promote the differentiation of T and B lymphocytes. Adenovirus AdV2 (adjuvant), without ligands, in combination with rRBD, served as a control. In vitro T-cell responses to the AdV1+rRBD combination revealed that CD8+ platform-specific T-cells increased (37.2 ± 0.7% vs. 23.1 ± 2.1%), and T-cells acted against SARS-CoV-2 via CD8+TEMRA (50.0 ± 1.3% vs. 36.0 ± 3.2%). Memory B cells were induced after treatment with either AdV1+rRBD (84.1 ± 0.8% vs. 82.3 ± 0.4%) or rRBD (94.6 ± 0.3% vs. 82.3 ± 0.4%). Class-switching from IgM and IgD to isotype IgG following induction with rRBD+Ab was observed. RNA-seq profiling identified gene expression patterns related to T helper cell differentiation that protect against pathogens. The analysis determined signaling pathways controlling the induction of protective immunity, including the MAPK cascade, adipocytokine, cAMP, TNF, and Toll-like receptor signaling pathway. The AdV1+rRBD formulation induced IL-6, IL-8, and TNF. RNA-seq of the VERO E6 cell line showed differences in the apoptosis gene expression stimulated with the platforms vs. mock. In conclusion, AdV1+rRBD effectively generates T and B memory cell-mediated protection, presenting promising results in producing CD8+ platform-specific T cells and isotype-switched IgG memory B cells. The platform induces protective immunity by controlling the Th1, Th2, and Th17 cell differentiation gene expression patterns. Further studies are required to confirm its effectiveness.

Constant exposure to various environmental and endogenous stresses can cause structural DNA damage, resulting in genome instability. Higher eukaryotic cells deploy conserved DNA repair systems, which include various DNA repair pathways, to maintain genome stability. Homologous recombination (HR), one of these repair pathways, involves multiple proteins. BRCA2, one of the proteins in the HR pathway, is of substantial research interest in humans because it is an oncogene. However, the study of this gene is limited due to the lack of availability of homozygous BRCA2-knockout mutants in mammals, which results in embryonic lethality. Arabidopsis thaliana has two copies of the BRCA2 homologs: BRCA2A and BRCA2B . Therefore, the single mutants remain nonlethal and fertile in Arabidopsis. The BRCA2A homolog, which plays a significant role in the HR pathway of germline cells and during the defense response, is well-studied in Arabidopsis. Our study focuses on the functional characterization of the BRCA2B homolog in the somatic cells of Arabidopsis, using the homozygous ΔBRCA2B mutant line. The phenotypic differences of ΔBRCA2B mutants were characterized and compared with wild Arabidopsis plants. The role of BRCA2B in spontaneous somatic HR (SHR) was studied using the ΔBRCA2B-gus detector line. ΔBRCA2B plants have a 6.3-fold lower SHR frequency than the control detector plants. Expression of four other HR pathway genes, including BRE, BRCC36A, RAD50, and RAD54, was significantly reduced in ΔBRCA2B mutants. Thus, our findings convey that the BRCA2B homolog plays an important role in maintaining spontaneous SHR rates and has a direct or indirect regulatory effect on the expression of other HR-related genes.

The aim of this study was to investigate the effect of Roundup^® herbicide on the maturation of Prussian carp oocytes under laboratory conditions. The Prussian carp is currently one of the most common fish species in Polish freshwater ichthyofauna. For the investigation, oocytes from five sexually mature female Prussian carp were used, segmented into three groups, and incubated for 24 h in Cortland's saline, treated with varied concentrations of the herbicide Roundup^® (0 ng - control, R1 - 10 ng/ml, and R2 - 100 ng/ml). Subsequent to this period, assays were performed using the prepared plates to determine the level of 17α,20β-dihydroxyprogesterone (17α,20β-P) utilizing the standard ELISA technique. In determining the 17α,20β-P via ELISA, the medium was extracted from each tested oocyte group. Oocyte maturity was assessed through preservation in serra fluid, and, to categorize the maturity stage of the oocytes utilizing a four-point scale - contingent upon the nucleus's position - the formerly preserved oocytes were dehydrated and subsequently analyzed. A contrast was noted in the percentage of oocytes at varied stages between the control group and the experimental groups. Specifically, a higher concentration of Roundup^® (100 ng/ml) accelerated to expedite the initial migration of the nucleus in oocytes. In conclusion, the obtained results show the adverse effect of Roundup^® on hormonal regulation and maturation in Prussian carp oocytes.

Essential oil (EO) from Eucalyptus polybrachtea is used as complementary and traditional medicine worldwide. The present study aimed at compositional profiling of EO and molecular docking of EO's bioactive compound 1,8 cineole against fungal enzymes involved in the riboflavin synthesis pathway, namely riboflavin synthase (RS), riboflavin biosynthesis protein RibD domain-containing protein (RibD), and 3,4-dihydroxy-2-butanone 4-phosphate synthase (DBPS) as apposite sites for drug designing against aspergillosis and mucormycosis, and in vitro confirmation. The compositional profile of EO was completed by GC-FID analysis. For molecular docking, the Patchdock tool was used. The ligand-enzyme 3-D interactions were examined, and ADMET properties (absorption, distribution, metabolism, excretion, and toxicity) were calculated. GC-FID discovered the occurrence of 1,8 cineole as a major component in EO, which was subsequently used for docking analysis. The docking analysis revealed that 1,8 cineole actively bound to RS, RibD, and DBPS fungal enzymes. The results of the docking studies demonstrated that the ligand 1,8 cineole exhibited H-bond and hydrophobic interactions with RS, RibD, and DBPS fungal enzymes. 1,8 cineole obeyed Lpinsky's rule and exhibited adequate bioactivity. Wet-lab authentication was achieved by using three fungal strains: Aspergillus niger, Aspergillus oryzae, and Mucor sp. Wet lab results indicated that EO was able to inhibit fungal growth.

Nanotechnology holds significance in all fields of research, and the formation and surface alterations of nanomaterials are particularly important in this discipline. Nanoformulations synthesized with bioactive plant components play a crucial role in the improvement of several therapeutics and diagnostics. In the present study, we reported the synthesis of a curcumin nanoformulation (CN) by using curcumin and D-α-tocopheryl polyethylene glycol 1000 succinate (TPGS). The synthesized CN was characterized using dynamic light scattering, UV-Visible spectrophotometry, Fourier-transform infrared spectroscopy, field-emission scanning electron microscopy, and X-ray diffraction. Furthermore, it was evaluated for solubility, drug loading, encapsulation efficiency, stability, in vitro release, and anticancer potentials. The role of TPGS in the synthesis of CN was validated. The synthesized CN exhibited a size of 6.2 ± 1.9 nm, needle-shaped morphology, a polydispersity index of 0.164, and zeta potential of - 10.1 ± 3.21 mV, as determined by characterization techniques. Its water solubility was 2.5 × 10⁴ times higher than that of pure curcumin. The encapsulation efficiency and curcumin loading efficiency of the synthesized CN were found to be 80 and 10%, respectively, with storage stability exceeding 30 days. Moreover, the synthesized CN demonstrated significant in vitro anticancer activity against the colorectal cancer cell line HCT-116, with an IC50 value of 12.74 ± 0.54 μM at 24 h.

Canthaxanthin is an orange-red keto-carotenoid that occurs naturally and is also manufactured by synthetic methods for regular applications. In nature, canthaxanthin mainly exists in microbes such as different bacterial species, fungi, and algae, as well as in animals such as crustaceans, certain fishes, and birds. However, the amount of canthaxanthin produced in these organisms varies significantly. Additionally, the compound can be generated from genetically modified organisms using genetic engineering techniques Canthaxanthin finds extensive application as an additive in animal feed, in the pharmaceutical industry, as a coloring agent for various food products, and in cosmetics. It has powerful antioxidant properties and plays a role in lipid metabolism, neuroprotection, and immunomodulation. This article gives an extensive insight into the structure and methods of synthesis of canthaxanthin along with its various newly discovered sources identified so far. The significant applications of canthaxanthin, particularly its role in pharmaceuticals, are critically evaluated. Furthermore, the article discusses future aspects and challenges associated with canthaxanthin production and regulation.

Clonal micropropagation is an effective method for plant reproduction, applicable in both scientific and industrial domains. However, a significant number of microclones are lost during the ex vitro acclimatization process. To address this, the introduction of beneficial microorganisms into the rhizosphere of micropropagated plants could have a positive effect on the survival rates and external characteristics of acclimatized plantlets. The aim of this study was to determine the protective and growth-promoting potential of Enterococcus italicus ONU547 and its effect on micropropagated plants during acclimatization. The antagonistic activity of the bacteria was determined using the agar block method. Lepidium sativum L. seeds were inoculated with bacterial suspensions at concentrations of 10⁶, 10⁷, and 10⁸ CFU/ml. Subsequently, the roots of the microclones were treated with suspensions of 10⁶ and 10⁷ CFU/ml, and biometric characteristics were measured. The results demonstrated antagonistic properties against various phytopathogenic fungi, including Aspergillus niger, Cladosporium cladosporioides, Alternaria alternata, Alternaria tenuissima, Rhizoctonia cerealis, Penicillium expansum, and Paecilomyces variotii. Inoculation of L. sativum L. seeds resulted in improved germination rates, increased root numbers, and enhanced root and shoot lengths. Similarly, the effects of the studied bacteria on Rubus fruticosus L. and Paulownia tomentosa Steud. during the acclimatization stage led to higher survival rates, increased shoot lengths, greater node numbers, and larger leaf areas. A concentration of 10⁷ CFU/ml was identified as optimal for inoculating the microclones. The findings indicate that E. italicus ONU547 holds promise for the inoculation of micropropagated plants during the acclimatization process. Further research is recommended to establish the specific interaction mechanisms between these bacteria and plants.

Chickpea is an important food legume cultivated in semiarid regions, where water scarcity and nutrient deficiencies negatively affect crop production. This study aimed to investigate the effect of zinc and silicon from different sources, including bulk and nanostructures, on various biochemical traits of chickpea plants grown under field conditions in Maragheh, Northwest Iran. The main experimental factor consisted of three soil moisture levels: irrigation to 90% of field capacity (FC), 60% FC, and 30% FC. The subplots were assigned for foliar application of different fertilizers: control (distilled water), zinc sulfate (ZnSO), silicon dioxide nanoparticles (SiO₂ NPs), ZnSO + SiO₂ NPs, and zinc-containing mesoporous silica nanoparticles (MSNPs -Zn). The results showed that although decreased soil moisture had a negative impact on several biochemical processes, foliar application of Zn and Si in both conventional bulk and nanostructure significantly affected plant antioxidant system, plasma membrane integrity, and the concentrations of photosynthetic pigments and compatible solutes. However, the most inducing effects on catalase, ascorbate peroxidase, guaiacol peroxidase, superoxide dismutase, and anthocyanin were observed with the foliar spray of MSNPs-Zn and ZnSO + SiO₂ under 60% FC. Moreover, foliar spray of MSNPs-Zn alleviated the negative effects of water deficit stress on photosynthetic pigments (chlorophyll a /b and carotenoid content). Water stress significantly induced the accumulation of free proline in the leaves. Overall, the results indicated that foliar spray of MSNPs -Zn, especially under 60% FC, improved the plant's defense system, scavenged reactive oxygen species, and enhanced the accumulation and stability of pigments, thereby mitigating the effects of drought stress.