3 Biotech最新文献

Animal diseases are among the most debilitating issues in the animal farming industry, resulting in decreased productivity and product quality worldwide. An emerging alternative to conventional injectable vaccines is edible vaccines, which promise increased delivery efficiency while maintaining vaccine effectiveness. One of the most promising platforms for edible vaccines is duckweeds, due to their high growth rate, ease of transformation, and excellent nutritional content. This review explores the potential, feasibility, and advantages of using duckweeds as platforms for edible vaccines. Duckweeds have proven to be superb feed sources, as evidenced by numerous improvements in both quantity (e.g., weight gain) and quality (e.g., yolk pigmentation). In terms of heterologous protein production, duckweeds, being plants, are capable of expressing proteins with complex structures and post-translational modifications. Research efforts have focused on the development of duckweed-based edible vaccines, including those against avian influenza, tuberculosis, Newcastle disease, and mastitis, among others. As with any emerging technology, the development of duckweeds as a platform for edible vaccines is still in its early stages compared to well-established injectable vaccines. It is evident that more proof-of-concept studies are required to bring edible vaccines closer to the current standards of conventional vaccines. Specifically, the duckweed expression system needs further development in areas such as yield and growth rate, especially when compared to bacterial and mammalian expression systems. Continued efforts in this field could lead to breakthroughs that significantly improve the resilience of the animal farming industry against disease threats.

Whole-cell bacteria overexpressing a combo of enzymes capable of breaking down complex lignocellulosic components of cell wall is a path-breaking innovation that is eco-friendly for agricultural waste processing and sustainable environment. In this study, a whole-cell E. coli overexpressing the enzyme alpha-galactosidase is used to biodegrade sugarcane bagasse, presenting a sustainable approach for agricultural waste utilization. Alpha-galactosidase is an enzyme that breaks down alpha-D-galactose residues at the non-reducing ends of oligosaccharides (such as raffinose, stachyose, and verbascose), complex galactomannans, and galactolipids. Submerged and solid-state fermentation-mediated hydrolysis of bagasse waste using recombinant E. coli overexpressing α-galactosidase shows a decrease in the level of α-galactosides releasing sucrose and reducing sugars, indicating a continuous breakdown of the cell wall. Scanning electron microscopy indicates substantial disintegration of cell wall fibers under both submerged (12 h) and solid-state (7 days) fermentation, confirming the disruption of bagasse cell wall structural integrity. The 2XM9 media was found competent for both total protein and enzyme activity; the total protein concentration was 2553 µg/ml after 28 h of induction with an enzyme activity of 0.445 gal units/µg of protein after 16 h of induction at 24 °C. The results show that using whole-cell recombinant systems that express different cell wall-degrading enzymes could be a sustainable way to use agricultural waste, which would help with both waste management and protecting the environment.

In this study, a novel colorimetric screening method for identifying menaquinone-7 (MK-7) producing strains was established using potassium permanganate. To our knowledge, this method represents the first direct screening methodology for the identification of MK-7 producing strains. Utilizing this screening method, a new MK-7 producing strain, Bacillus subtilis GSA-184, was identified from the soil of the Tibetan Plateau. Under the optimized fermentation medium (50 g/L glycerol, 30 g/L yeast extract powder, 100 g/L soybean peptone, 1 g/L KH₂PO₄, and 1 g/L MnSO₄), the production of MK-7 was increased to 25.7 mg/L. Additionally, the maximum production of MK-7 reached 36.46 mg/L after 48 h in a 5-L fermenter.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-024-04097-1.

Hydrothermal pretreatments are commonly employed prior to the biotechnological conversion of lignocellulosic biomass (LCB) into value-added products, such as fuels and chemicals. However, the by-products of this pretreatment, including furaldehydes, lignin-derived phenolics, and carboxylic acids, can inhibit the enzymes and microbes used in the biotechnological process. In this study, LCB degrading enzymes of endophytic and litter fungi were screened for their tolerance to potential pretreatment-derived inhibitors. Several fungi produced endo- and exoglucanases that remained functional in the presence of lignocellulose-derived phenolics. Some were also active in the presence of tannic acid. Additionally, thermostable endoglucanase activity was observed in some fungi. The ability of some of these fungi to utilize furaldehyde inhibitors as a sole carbon source was also noted. The culture supernatants of the fungal strains were tested in hydrolysis experiments using microcrystalline cellulose as a substrate, in the presence of lignocellulose phenolics and tannic acid. With some strains, higher sugar yields were obtained in the hydrolysis of cellulose when phenolics were added. Our results highlight the need for more intensive exploration of endophytic and plant litter fungi for novel inhibitor-resistant cellulases for biofuel production.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-024-04087-3.

The current investigation involved the isolation of 13 endophytic fungi from Taxus sp. collected in Himachal Pradesh, India. Among these, isolate PAT-3 produced 195.13 mg/L of Taxol in reformative medium broth using microbial fermentation as an alternative source. The PAT-3 isolate was characterized as Mucor circinelloides through morphologic and molecular techniques. The PAT-3 isolate was the only one to exhibit positive results for the Taxol biosynthesis-related genes 10-deacetylbaccatin-III-10-O-acetyltransferase (dbat), Baccatin-III, 3: amino, 3 phenylpropanol transferase (bapt), and taxadienol-acetyltransferase (tat). Furthermore, human breast cancer (MCF-7) and human melanoma cancer (SKMEL-28) cell lines demonstrated the cytotoxicity of Taxol extracted from isolate PAT-3, with IC₅₀ values of 80.32 µg/mL and 77.21 µg/mL, respectively. To our knowledge, this is the first study that demonstrates the ability of the endophytic fungus M. circinelloides from Taxus sp. in the northern Himalayan region to produce paclitaxel. The study's findings show that Mucor circinelloides is an excellent alternative source of Taxol, and they may pave the way for the production of Taxol at the industrial level in future.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-024-04091-7.

Anamirta cocculus, a woody climber, is extensively utilised in traditional Asian medicine. This study investigates the cytotoxic effects of A. cocculus leaf extracts on various cancer cell lines as well as on a normal cell line. The ethyl acetate extract exhibited potent anticancer activity, with the highest cytotoxicity observed against ovarian cancer cell line (PA1) (IC₅₀ = 8.30 ± 0.38 µg/mL) and colorectal adenocarcinoma cell line (HT29) (IC₅₀ = 17.97 ± 0.63 µg/mL). Notably, the extract displayed low toxicity (18.72 ± 0.73%) on the normal human keratinocyte cell line (HaCaT) at a concentration of 100 µg/mL, indicating selective cytotoxicity towards cancer cells. The acetone extract also demonstrated significant cytotoxicity against various cancer cell lines, including A498, MG63, PA1, and UM-SCC-83B. The ethyl acetate extract of A. cocculus demonstrated potent inhibition of colony formation in HT29 and PA1 cancer cell lines while inducing apoptosis, as evidenced by membrane blebbing, chromatin condensation, and DNA fragmentation. The number of late apoptotic cells increased with an increase in concentrations of ACLE. Molecular docking studies of compounds identified through GC-MS analysis revealed strong interactions with key apoptotic proteins, including caspase-8, p53, caspase-3, and caspase-9. Compounds such as vitamin E, epoxylathyrol, squalene, and phytol showed high binding affinity to these proteins, suggesting their role in apoptosis induction. The possibility of induction of apoptotic proteins through indirect interaction by binding to other proteins or receptors cannot be ruled out. The cytotoxic effects may result from individual, combined, or synergistic actions of these compounds. Among these, epoxylathyrol emerged as a particularly promising anticancer drug candidate based on ADME analysis and binding affinity assessments, warranting further investigation.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-024-04096-2.

The rapid rise of multidrug-resistant (MDR) organisms has created a critical need for alternative treatment options. Phage therapy is gaining attention as an effective way to fight bacterial infections by using lytic bacteriophages to specifically target and kill harmful bacteria. This review discusses several phage therapeutic options and emphasizes new developments in phage biology. Phage treatment has proven to be successful against MDR bacteria, as evidenced by multiple human clinical trials that indicate favorable results in treating a range of diseases caused by these pathogens. Despite these promising results, challenges such as phage resistance, regulatory hurdles, and the need for standardized treatment protocols remain. To effectively combat MDR bacterial infections, future research must focus on enhancing phage effectiveness, guaranteeing safety for human usage and incorporating phage therapy into clinical practice.

In the current study, the ability of an indigenous marine Actinomycete Nocardiopsis dassonvillei (NCIM 5124) to degrade poly(3-hydroxybutyrate)-PHB was examined. From the whole genome sequencing data of the organism, information regarding the PHB depolymerase gene and amino acid sequence (Accession number: MCK9871921.1) was retrieved. In silico studies indicated the presence of a signal peptide characteristic of extracellular enzymes. ProtParam tool predicted that the protein had a molecular mass of 42.46 kDa with an isoelectric point of 4.51. Aliphatic and instability index values suggested that the protein was stable and the observed GARVY value indicated its hydrophilic nature. 3D structure prediction and multiple sequence alignments revealed the presence of Type I catalytic domain [including the oxyanion histidine towards the N terminal, the catalytic triad with serine (as a part of GLSAG pentapeptide), aspartate and histidine], substrate binding and linker domain. The organism was able to grow on PHB in solid and liquid media and effectively degrade it. Maximum enzyme activity (1.8 U/mL/min) was observed after 5 d of incubation in Bushnell Hass Medium containing 0.1% PHB, 1.5% sodium chloride, at 30 °C, pH 7.5 with agitation at 130 rpm. Application of the organism in disintegrating films of PHB and its copolymers was successfully demonstrated on the basis of weight loss and scanning electron microscope analysis. To the best of our knowledge, this is the first report on production of PHB depolymerase with high efficiency by N. dassonvillei, an organism that holds promise in degrading PHB-derived waste material.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-024-04079-3.

This study investigates the response of ethyl methanesulfonate-derived twenty mutant lines of Gossypium herbaceum, along with the parent type Wagad cultivar, to drought stress. Physiological parameters, such as relative water content (RWC), net photosynthesis (A), stomatal conductance (g _s), transpiration rate (E), and water use efficiency (WUE), were examined. The mutant line mut_3219 exhibited superior drought tolerance, maintaining high RWC and water retention capacity, with minimal reductions in A, g _s, and E, leading to higher WUE than parent type and other mutant lines. Chlorophyll pigments declined in all the mutants under drought. However, mut_3219 retained higher levels than mut_4785. Anthocyanin accumulation indicated a protective response. Chlorophyll fluorescence showed mut_3219 is less sensitive to drought-induced PSII damage than mut_4785, with better membrane stability and higher proline accumulation, among all other mutant lines and parent type. The morphological parameters were less affected in mut_3219 compared to mut_4785 and parent type. Molecular analyses under control and drought conditions revealed significant variations in the expression of seven drought-related genes (GhbHLH, GhMYB5, GhWRKY33, GhRAF4, GhRAF19, GhNAC2, and GhCAMTA). The relative expression of GhbHLH, GhNAC2, GhRAF4, GhRAF19, and GhCAMTA increased under drought conditions, with notable changes in mut_3219 compared to parent type and all other mutant lines, indicating its enhanced drought tolerance. These findings provide valuable insights into the molecular and physiological mechanisms underlying drought tolerance in cotton.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-024-04089-1.

The present study was aimed to synthesize the silver nanoparticles from Alangium salvifolium Wang. and evaluating its biomedical applications. The leaves of A. salvifolium collected and subjected for the standard procedure of Soxhlet extraction using distilled water as a solvent. With the help of an aqueous extract AgNPs were synthesized from silver nitrate using phyto-reduction method. Further, synthesized AgNPs were characterized using several analytical techniques such as UV, FTIR, SEM-EDX, XRD, particles size and zeta potential. Synthesized AgNPs were tested for antibacterial, antioxidant, anticancer for lung cancer cell line and flowcytometry-based pathway studies. The visual observation confirmed the formation of AgNPs from the aqueous extract by changing yellow to brown colour formation. Further, characterization techniques also confirmed the formation of AgNPs. Antibacterial activity results showed that the tested AgNPs were potent against bacterial pathogens with a higher zone of inhibition. Further, the antioxidant and anticancer activity of AgNPs revealed that the AgNPs have exhibited significant results with a good percentage of inhibition. Further, the flow cytometry studies confirmed that the AgNPs inducing apoptosis and cell cycle arrest in lung cancer. The phytochemicals of A. salvifolium plant have successfully synthesized AgNPs. In the case of performed biological activity, the synthesized silver nanoparticles exhibited potent activity. In future these AgNPs can be taken for molecular and in vivo studies to identify their efficacy using in vivo and molecular models.