3 Biotech最新文献

Diabetic mellitus (DM) is characterized by hyperglycaemia and defective macromolecular metabolism, arising from insulin resistance or lack of insulin production. The present study investigates the potential of artemisinin, a sesquiterpene lactone isolated from Artemisia annua, to exert anti-diabetic and antioxidant effects through modulation of the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signalling pathway. Our computational analyses demonstrated a high binding affinity of artemisinin with proteins belonging to the PI3K/AKT signalling cascade. α-Amylase and α-glucosidase studies revealed a notable increase in inhibition percentages with artemisinin treatment across concentrations ranging from 10 to 160 µM. A similar significant (p < 0.05) dose-dependent inhibition of free radicals was observed for the in vitro anti-oxidant assays. Further, toxicological profiling of artemisinin in the in vivo zebrafish embryo-larvae model from 4 to 96 h post-fertilization (hpf) did not exhibit any harmful repercussions. In addition, gene expression investigations confirmed artemisinin's potential mechanism in modulating hyperglycaemia and oxidative stress through the regulation of the PI3K/AKT pathway. Overall, our investigation suggests that artemisinin can be used as a therapeutic intervention for diabetes and oxidative stress, opening up opportunities for future investigation in clinical settings.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-024-04050-2.

In the course of past two decade anthropogenic activities have reinforced, begetting soil and water defilement. A plethora of heavy metals alters and limits plant growth and yield, with opposing effect on agricultural productivity. Silicon often perceived as plant alimentary 'nonentity'. A suite of determinants associated with silicon have been lately discerned, concerning plant physiology, chemistry, gene regulation/expression and interaction with different organisms. Exogenous supplementation of silicon renders resistance against heavy-metal stress. Predominantly, plants having significant amount of silicon in root and shoot thus are barely prone to pest onset and manifest greater endurance against abiotic stresses including heavy-metal toxicity. Silicon-mediated stress management involves abatement of metal ions within soil, co-precipitation of metal ions, gene modulation associated with metal transport, chelation, activation of antioxidants (enzymatic and non-enzymatic), metal ion compartmentation and structural metamorphosis in plants. Silicon supplementation also stimulates expression of stress-resistant genes under heavy-metal toxicity to provide plant tolerance under stress conditions. Ergo, to boost metal tolerance within crops, immanent genetic potential for silicon assimilation should be enhanced. Current study, addresses the potential role and mechanistic interpretation of silicon induced mitigation of heavy-metal stress in plants.

Plant pathogens are causing substantial economic losses and thus became a significant threat to global agriculture. Effective and timely detection methods are prerequisite for combating the damages caused by the plant pathogens. In the realm of plant pathogen detection, the isothermal amplification techniques, e.g., recombinase polymerase amplification (RPA) and loop-mediated isothermal amplification (LAMP), have emerged as a fast, precise, and most sensitive alternative to conventional PCR but they often comprise high rates of non-specific amplification and operational complexity. In recent advancements, clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated nuclease Cas systems, particularly Cas12, have emerged as powerful tools for highly sensitive, specific, and rapid pathogen detection. Exploiting the collateral activities of Cas12, which selectively cleaves single-stranded DNA (ssDNA), novel detection platforms have been developed. The mechanism employs the formation of a triple complex molecule comprising guide RNA, Cas12 enzyme, and the substrate target nucleotide sequence. Upon recognition of the target, Cas12 indiscriminately cleaves the DNA strand, leading to the release of fluorescence from the cleaved ssDNA reporter. Integration of isothermal amplification methods with CRISPR/Cas12 enables one-step detection assays, facilitating rapid pathogen identification within 30 min at a single temperature. This integrated RPA-CRISPR/Cas12a approach eliminates the need for RNA extraction and cDNA conversion, allowing direct use of crude plant sap as a template. With an affordable fluorescence visualization system, this portable method achieves 100-fold greater sensitivity than conventional techniques. This review summarizes recent advances in RPA-CRISPR/Cas12a for detecting plant pathogens, covering primer design, field-level portability, and enhanced sensitivity.

This study aimed to explore the protective mechanism of Banxia Xiexin Tang (BXXXT) on liver cell damage caused by high glucose (H-G) and to clarify its molecular regulatory pathways. First, the main components in BXXXT-containing serum were analyzed by high-performance liquid chromatography (HPLC) to provide basic data for subsequent experiments. Subsequently, the effect of BXXXT on high glucose (H-G)-induced hepatocyte activity was evaluated through screening of the optimal concentration of drug-containing serum. Experimental results showed that BXXXT significantly reduced the loss of cell activity caused by high glucose. Further research focuses on the regulatory effect of BXXXT on high glucose-induced hepatocyte apoptosis, especially its effect on the PGC-1α (peroxisome proliferator-activated receptor γ coactivator-1α) pathway. Experimental results showed that BXXXT reduced high-glucose-induced hepatocyte apoptosis and exerted its protective effect by upregulating the activity of the PGC-1α pathway. BXXXT significantly increased the expression level of IGFBP1 (insulin-like growth factor-binding proteins) in hepatocytes under a high-glucose environment. It cleared mitochondrial ROS (reactive oxygen species) by enhancing SOD2 (superoxide dismutase) enzyme activity and maintained the survival of hepatocytes under a high-glucose environment. Finally, the regulation of PGC-1α by BXXXT is indeed involved in the regulation of IGFBP1 expression in hepatocytes and its downstream SOD2 effector signaling. Taken together, this study provides an in-depth explanation of the protective mechanism of BXXXT on hepatocytes in a high-glucose environment, focusing on regulating the expression of the PGC-1α pathway and IGFBP1, and reducing cell damage by scavenging ROS. This provides an experimental basis for further exploring the potential of BXXXT in the treatment of diabetes-related liver injury.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-024-04060-0.

The present study examines the use of waste cooking oil (WCO) as a substrate for medium-chain-length polyhydroxyalkanoates (mcl-PHA) production by Pseudomonas rhizophila S211. The genome analysis revealed that the S211 strain has a mcl-PHA cluster (phaC1ZC2DFI) encoding two class II PHA synthases (PhaC1 and PhaC2) separated by a PHA depolymerase (PhaZ), a transcriptional activator (PhaD) and two phasin-like proteins (PhaFI). Genomic annotation also identified a gene encoding family I.3 lipase that was able to hydrolyze plant oils and generate fatty acids as favorable carbon sources for cell growth and PHA synthesis via β-oxidation pathway. Using a three-variable Doehlert experimental design, the optimum conditions for mcl-PHA accumulation were achieved in 10% of WCO-based medium with an inoculum size of 10% and an incubation period of 48 h at 30 °C. The experimental yield of PHA from WCO was 1.8 g/L close to the predicted yield of 1.68 ± 0.14 g/L. Moreover, ¹H nuclear magnetic resonance spectroscopy analysis confirmed the extracted mcl-PHA. Overall, this study describes P. rhizophila as a cell factory for biosynthesis of biodegradable plastics and proposes green and efficient approach to cooking oil waste management by decreasing the cost of mcl-PHA production, which can help reduce the dependence on petroleum-based plastics.

Phosphorus (P) is the key to several structural molecules and catalyzes numerous biochemical reactions in plant body besides its involvement in energy transfer. Any deficit in P availability is likely to result in reduced RNA and protein content, inhibiting crop growth and development. Thus, availability of soil P is extremely crucial for plant growth especially in acid soils of India, where most of the fraction is bound to solid phase rendering their availability. The present communication deals with the isolation of elite phosphate-solubilizing bacterial (PSB) strains from the acid soils to work out their ability to improve the fertilizer P use efficiency in the acidic environment. Initially twenty-six bacteria were isolated from the acid soils of Northeastern India. Among them, ten bacteria were selected based on formation of halo zone in the Pikovskaya agar plate. In addition, these bacteria were able to solubilize insoluble zinc (Zn) and potassium (K). The isolates were subject to in vitro optimization for P solubilization under different insoluble P source utilization and at different pH and salinity conditions. Strains AN3, AN11, and AN21 exhibited significant solubilization of insoluble P, Zn, and K, and were identified as Streptomyces sp., Enterobacter sp., and Paraburkholderia caribensis. These three bacteria solubilized 206.53 to 254.08 µg mL^-1 P, 79.7 to 177.55 µg mL^-1 Zn, and 0.96 to 1.56 µg mL^-1 K from insoluble minerals. Their performance was further evaluated in pot culture experiment using green gram as test crop. These three bacteria were found to improve P uptake and dry matter accumulation in green gram plant substantially. Seed bio-priming with the PSB strains enhanced the efficiency of added P fertilizer, resulting in a 1.40 to 1.52 times higher effectiveness compared to the control. On the whole, AN11 may be ranked as best inoculant for the acidic soils of Northeastern India.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-024-04042-2.

The present study investigated antiviral peptides (AVPs) as inhibitors of SARS-CoV-2 Orf9b protein, a novel target for disrupting the Orf9b-TOM70 complex crucial for viral infection. In silico screening via molecular docking and MD simulations identified AVP1442 and AVP1896 with high binding affinities to Orf9b (- 846.3 kcal mol^-1 and - 820 kcal mol^-1, respectively), comparable to the Orf9b-TOM70 complex (- 810.99 kcal mol^-1). These AVPs interacted with key amino acid residues in Orf9b, including phosphorylation sites. In addition, AVPs also closely interacted with conserved regions in Orf9b. AVP1896 formed a hydrogen bond with Orf9b's threonine at position 84. AVP1442 interacted with Orf9b's leucine at position 15. Favorable Ramachandran plots and compactness during MD simulations for up to 100 ns suggest good stability of formed complexes. These non-toxic AVPs warrant further in vitro and in vivo evaluation, potentially as components of drug cocktails with small molecules or interferon-based therapies.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-024-04032-4.

Komagataella phaffii (previously described as Pichia pastoris) is a yeast that produces high-level heterologous proteins with a wide range of applications in medicine and industry. The methanol-induced alcohol oxidase I promoter (P_AOX1) is frequently used for protein expression in this yeast. However, limitations on the use of methanol have been observed in large-scale production, including its flammability, toxicity, and need for special handling. Here, we propose to develop a system using recombinant cells constitutively expressing pectinmethyl esterase for expression of two reporter proteins, GFP and azurin, under the control of P_AOX1 using pectin in production medium. So, this system is coherent with yeast culture medium containing pectin and heterologous gene inserted downstream of P_AOX1 can be successfully expressed without the addition of methanol. Therefore, this novel Self-inducibLe heterologous protein EXpression (SILEX) system, which does not require the addition of methanol, can be used for the production of any protein. It can also be adapted for large-scale production.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-024-04039-x.

Azo dye-laden textile wastewater must be treated before release due to various health and environmental concerns. Bioremediation of textile wastewater, however, is a challenge owing to its alkaline and saline nature as mesophilic microbes, in general, are either not able to thrive or show less efficiency under such hostile environment. Thus, pre-treatment for neutralization or salinity removal becomes a prerequisite before applying microbes for treatment, causing extra economical and technical burden. Extremophilic bacteria can be the promising bioremediating tool because of their inherent ability to survive and show toxicants removal capability under such extreme conditions without need of pre-treatment. Among extremophiles, halophilic and alkaliphilic bacteria which are naturally adapted to high salt and pH are of special interest for the decolorization of saline-alkaline-rich textile wastewater. The current review article is an attempt to provide an overview of the bioremediation of azo dyes and azo dye-laden textile wastewater using these two classes of extremophilic bacteria. The harmful effects of azo dyes on human health and environment have been discussed herein. Halo-alkaliphilic bacteria circumvent the extreme conditions by various adaptations, e.g., production of certain enzymes, adjustment at the protein level, pH homeostasis, and other structural adaptations that have been highlighted in this review. The unique properties of alkaliphiles and halophiles, to not only sustain but also harboring high dye removal competence at high pH and salt concentration, make them a good candidate for designing future bioremediation strategies for the management of alkaline, salt, and azo dye-laden industrial wastewaters.

Considering the current growing interest in new and improved enzymes for use in a variety of applications, the present study aimed to characterize a novel detergent-stable serine alkaline protease from the extremophilic actinobacterium Microbacterium metallidurans TL13 (MmSP) using a combined in silico and experimental approach. The MmSP showed a close phylogenetic relationship with high molecular weight S8 peptidases of Microbacterium species. Moreover, its physical and chemical parameters computed using Expasy’s ProtParam tool revealed that MmSP is hydrophilic, halophilic and thermo-alkali stable. 3D structure modelling and functional prediction of TL13 serine protease resulted in the detection of five characteristic domains: [catalytic subtilase domain, fibronectin (Fn) type-III domain, peptidase inhibitor I9, protease-associated (PA) domain and bacterial Ig-like domain (group 3)], as well as the three amino acid residues [aspartate (D182), histidine (H272) and serine (S604)] in the catalytic subtilase domain. The extremophilic strain TL13 was tested for protease production using agricultural wastes/by-products as carbon substrates. Maximum enzyme activity (390 U/gds) was obtained at 8th day fermentation on potato peel medium. Extracellular extract was concentrated and partially purified using ammonium sulfate precipitation methodology (1.58 folds purification fold). The optimal pH, temperature and salinity of MmSP were 9, 60 °C and 1 M NaCl, respectively. The MmSP protease showed broad pH stability, thermal stability, salt tolerance and detergent compatibility. In order to achieve the maximum stain removal efficacy by the TL 13 serine protease, the operation conditions were optimized using a Box–Behnken Design (BBD) with four variables, namely, time (15–75 min), temperature (30–60 °C), MmSP enzyme concentration (5–10 U/mL) and pH (7–11). The maximum stain removal yield (95 ± 4%) obtained under the optimal enzymatic operation conditions (treatment with 7.5 U/mL of MmSP during 30 min at 32 °C and pH9) was in good agreement with the value predicted by the regression model (98 ± %), which prove the validity of the fitted model. In conclusion, MmSP appears to be a good candidate for industrial applications, particularly in laundry detergent formulations, due to its high hydrophilicity, alkali-halo-stability, detergent compatibility and stain removal efficiency.